ClovisFerreiraDosReis TESE

UNIVERSIDADE FEDERAL DO RIO GRANDE DO NORTE
INSTITUTO METRÓPOLE DIGITAL

PROGRAMA DE PÓS-GRADUAÇÃO EM BIOINFORMÁTICA
DOUTORADO EM BIOINFORMÁTICA
Clovis Ferreira dos Reis
Análise Baseada em Biologia de Sistemas de

Dados Transcricionais de Células Progenitoras
Neurais Humanas Tratadas com Chumbo
Natal - RN
2019
Clovis Ferreira dos Reis
Análise Baseada em Biologia de Sistemas de Dados

Transcricionais de Células Progenitoras Neurais Humanas
Tratadas com Chumbo
Orientador: Prof. Dr. Rodrigo Juliani Siqueira Dalmolin – UFRN

Coorientador: Profa . Dra . Rita Maria Cunha de Almeida – UFRGS
Natal - RN
2019
Análise Baseada em Biologia de Sistemas de Dados

Transcricionais de Células Progenitoras Neurais Humanas
Tratadas com Chumbo
Tese apresentada ao Programa de Pós-graduação

em BioInformática da Universidade Federal do Rio
Grande do Norte por Clovis Ferreira dos Reis como
parte dos requisitos para a obtenção do tı́tulo de
Doutor em BioInformática.
Área de concentração: Bioinformática
Linha de Pesquisa: Genômica
Orientador: Prof. Dr. Rodrigo Juliani S. Dalmolin

Coorientadora: Profa . Dra . Rita Maria C. de
Almeida
Natal, RN
2019
Universidade Federal do Rio Grande do Norte - UFRN
Sistema de Bibliotecas - SISBI
Catalogação de Publicação na Fonte. UFRN - Biblioteca Central Zila Mamede
Reis, Clovis Ferreira dos.

Análise baseada em biologia de sistemas de dados
transcricionais de células progenitoras neurais humanas tratadas
com chumbo / Clovis Ferreira dos Reis. - 2019.
112f.: il.
Tese (Doutorado)-Universidade Federal do Rio Grande do Norte,

Instituto Metrópole Digital, Programa de Pós-Graduação em
Bioinformática, Natal, 2019.
Orientador: Dr. Rodrigo Juliani Siqueira Dalmolin.
Coorientadora: Dra. Rita Maria Cunha de Almeida.
1. Exposição ao chumbo - Tese. 2. Análise de transcriptoma -

Tese. 3. Inferência de redes - Tese. 4. Visualização de redes -
Tese. 5. Transcriptogramer - Tese. I. Dalmolin, Rodrigo Juliani
Siqueira. II. Almeida, Rita Maria Cunha de. III. Título.
RN/UF/BCZM CDU 004:577
Elaborado por Raimundo Muniz de Oliveira - CRB-15/429

Agradecimentos
Aos colegas do BioME pela convivência;

Aos professores pelo conhecimento;
Aos colegas de pesquisa Iara, Raffael, Diego e Danilo pela colaboração;
Aos orientadores Rodrigo e Rita pela inspiração;
Aos meus irmãos Jeane, Maristela, Celso, Jairo e Milene pelo afeto;
À Ivana, mesmo à distancia, pela companhia;
Aos meus filhos Junior, Victor e Larissa pelo meu legado;
Aos meus pais Euclydes e Maria das Graças pela existência; e
A todos estes pela amizade.
Sumário
Lista de ilustrações . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Lista de tabelas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Introdução 13
1 CONSIDERAÇÕES SOBRE O CHUMBO . . . . . . . . . . . . . . . 13

1.1 O chumbo como uma questão de saúde pública . . . . . . . . . . . . 13
1.2 Efeitos neurotóxicos do chumbo . . . . . . . . . . . . . . . . . . . . . 15
1.3 O impacto sistêmico do envenenamento por chumbo . . . . . . . . . 18
2 DIFERENCIAÇÃO CELULAR . . . . . . . . . . . . . . . . . . . . . 20
2.1 Caracterizando a diferenciação neuronal em um transcriptoma . . . 21
3 ANÁLISE DO TRANSCRIPTOGRAMA . . . . . . . . . . . . . . . . 24
3.1 O pacote transcriptogramer . . . . . . . . . . . . . . . . . . . . . . . . 24
Justificativa 30
Objetivos 31
Artigo 33
Discussão 44
Conclusão 53
REFERÊNCIAS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
APÊNDICE A – MATERIAL SUPLEMENTAR DO ARTIGO . . . . 61
6
Lista de ilustrações
Figura 1 – Número de casos de intoxicação por chumbo em crianças (USA) . . . . 15

Figura 2 – Resumo de vias metabólicas afetadas pelo cálcio . . . . . . . . . . . . . 18
Figura 3 – Modelo esquemático dos efeitos induzidos na célula . . . . . . . . . . . 19
Figura 4 – Modelo esquemático da diferenciação neuronal. . . . . . . . . . . . . . 21
Figura 5 – Matriz PPI de exemplo. . . . . . . . . . . . . . . . . . . . . . . . . . . 25
Figura 6 – Exemplo de ordenamento. . . . . . . . . . . . . . . . . . . . . . . . . . 25
Figura 7 – Exemplo de janela deslizante . . . . . . . . . . . . . . . . . . . . . . . 26
Figura 8 – Representação gráfica das médias de DE . . . . . . . . . . . . . . . . . 27
Figura 9 – Determinação dos clusteres funcionais . . . . . . . . . . . . . . . . . . 28
Figura 10 – Representação gráfica dos clusteres funcionais . . . . . . . . . . . . . . 28
Figura 11 – Rede hierárquica de termos de GO . . . . . . . . . . . . . . . . . . . . 29
Figura 12 – Interação Delta1/NOTCH1 . . . . . . . . . . . . . . . . . . . . . . . . 46
7
Lista de tabelas
Tabela 1 – Exemplo de enriquecimento de ontologias. . . . . . . . . . . . . . . . . 29
8
Lista de Sı́mbolos e Abreviações
A.C. – Antes de Cristo

CaM – Calmodulina
cAMP – Cyclic AMP
CaMK – CaM-kinases
CDC – US Center of Disease Control
CFM – Cost Function Minimizing
CNS – Sistema nervoso central (Central Nervous System)
DE – Expressão Diferencial
EDTA – Ácido etilenodiamino tetra-acético (Ethylenediamine tetraacetic acid)
ESC – Embryonic Stem Cells
ES-NP cells – Human embryonic-derived neural progenitor cells
Glu – Glutamato
GO – Gene Ontology
hCG – Gonadotrofina coriônica
Hb – Hemoglobina
HPT – Hipotálamo, Pituitária, Testı́culos
NGF – Nerve Growth Factor
NMDAR – Receptor N–methyl–D–aspartato
NPCs – Células progenitoras neurais humanas
PKC – Proteı́na Quinase C
PPI – Interação proteı́na proteı́na
T1 – Time-Interval 1
T2 – Time-Interval 2
9
Resumo
As consequências do envenenamento por chumbo são diversas e importantes na saúde
humana uma vez que este metal pesado pode interagir com muitos sistemas orgânicos,
afetando principalmente o sistema nervoso, com implicações graves e irreversı́veis do
neurodesenvolvimento, consolidação de memória e processos de aprendizagem em crianças.
Sua interação com componentes celulares dá-se de muitas formas, afetando proteı́nas de
ligação a ı́ons, proteı́nas de sinalização de transdução, canais iônicos transmembrana e
fatores de transcrição. Apesar da sintomatologia da intoxicação por chumbo já ser bastante
conhecida, pouco ainda se sabe sobre seus efeitos sistêmicos e sobre o seu impacto global
na modulação da transcrição de células neuronais. A fim de investigar tais efeitos sob uma
ótica de biologia de sistemas, aplicamos o pipeline do pacote transcriptogramer R/Biocon-
ductor com a finalidade de avaliar o perfil transcricional de células progenitoras neurais
humanas (NPCs) tratadas com acetato de chumbo 30µM por 26 dias. Dotado de um
método não supervisionado, o algoritmo do transcriptogramer é projetado para identificar,
em experimentos do tipo caso-controle, grupos de genes funcionalmente associados e difer-
encialmente expressos. Tal pipeline foi capaz de identificar onze clusteres diferencialmente
expressos entre os dias 3 e 11 do tratamento com chumbo. Destes, sete apresentaram
uma regulação negativa de diversos sistemas celulares envolvidos na diferenciação celular,
como organização do citoesqueleto, RNA e biossı́ntese de proteı́nas, caracterizados por
redes grandes e fortemente conectadas. Os quatro clusteres positivamente regulados
apresentaram nós esparsos e pouco conectados, principalmente relacionados a transcrição,
transporte transmembrana e transdução de sinal. Já no perı́odo subsequente, envolvendo
os dias 12 a 26 de tratamento, foi possı́vel observar uma alteração maciça do perfil de
transcrição celular com interferência em todas as camadas da regulação da expressão gênica.
Desta forma, nossos resultados sugerem que o chumbo induz modificações transcricionais
significativas nas NPCs que podem ser correlacionadas a danos e/ou adaptações de diversos
sistemas, todos decorrentes da intoxicação por este metal pesado, influenciando, assim, o
resultado final da diferenciação das células ES-NP.
Palavras-chaves: exposição ao chumbo, transcriptogramer, RNA-Seq, análise de tran-
scriptoma, time series, inferência de redes, data integration, visualização de redes.
10
Abstract
The consequences of lead poisoning are diverse and relevant to human health. Reaching all
organ systems, it mainly affects the nervous system, with severe and irreversible implications
of neurodevelopment, memory consolidation, and learning processes in children. They
interact with cellular components in many ways, affecting ion-binding proteins, transduction
signaling proteins, transmembrane ion channels, and transcription factors. If in one hand,
the symptoms of lead poisoning are well known, on the other hand, we have a lack of the
systemic effects and its impact on neuronal cell transcription modulation. In order to
investigate such effects from a systems biology perspective, we applied the transcriptogramer
R/Bioconductor package pipeline to evaluate the transcriptional profile of lead acetate-
treated human neural progenitor cells (NPCs) 30µM for 26 days. The transcriptogramer
algorithm is designed to identify functionally associated and differentially expressed gene
groups in case-control experiments in an unsupervised way. It was able to identify eleven
differentially expressed clusters between days 3 and 11 of the lead treatment. Of these,
seven presented negative regulation of several cellular systems involved in cell differentiation,
such as cytoskeleton organization, RNA and protein biosynthesis, characterized by large
and tightly connected networks. The four clusters that were positively regulated presented
sparse and poorly connected nodes, mainly related to transcription, transmembrane
transport, and signal transduction. In the subsequent period, involving days 12 to 26 of
treatment, it was possible to observe a massive alteration of the cellular transcription
profile with interference in all layers of gene expression regulation. Thus, our results
suggest that lead induces significant transcriptional modifications in NPCs which can be
correlated to damage and/or adaptations of various systems, all resulting from intoxication
by this heavy metal, thus influencing the result of ES-NP cell differentiation.
Key words: lead (Pb) exposure, transcriptogramer, RNA-Seq, transcriptome analysis,
time series, network inference, data integration, network visualization.
11
Introdução
13
1 Considerações sobre o chumbo
O chumbo é, provavelmente, um dos primeiros minerais que o homem veio a utilizar em
metalurgia, sendo o artefato mais antigo feito com este material datado em 4300–4000 A.C.
[1]. De sı́mbolo quı́mico P b, número atômico 82 e massa atômica igual a 207,2 daltons é
classificado como metal pesado. É o metal estável de maior número atômico e o sétimo metal
mais denso, com densidade de 11, 34g/cm3 , atrás da platina, tungstênio, ouro, urânio, mercúrio
e paládio, sendo entre estes o mais abundante. Apresenta valência variável, podendo assumir
as valências 2 e 4 (P b2+ e P b4+ , respectivamente).
Possui as seguintes caracterı́sticas fı́sico-quı́micas:
– Maleabilidade, podendo ser facilmente reduzido a lâminas finas;
– Ductibilidade, podendo deformar-se sem romper-se sob pressão cisalhante;
– Baixa condutividade elétrica;
– Baixo ponto de fusão, tornando-se lı́quido a 327,4o C e;
– Altamente resistente à corrosão.
Tais caracterı́sticas o tornam um material muito versátil e de fácil manipulação, fato este
que, aliado ao seu baixo custo de produção e abundância, o tornam um insumo importante
em diversas áreas econômicas, sendo o quinto metal mais consumido pela indústria, superado
apenas pelo ferro, cobre, alumı́nio e o zinco [2]. Devido às suas caracterı́sticas isolantes e
de resistência à oxidação, é largamente empregado na indústria elétrica, em acumuladores de
energia (baterias) e em forros para cabos elétricos. Possuindo elevada densidade, se presta
muito bem ao revestimento de ambientes como isolante acústico e eletromagnético, assim
como blindagem contra radiações de alta energia. Seu baixo ponto de fusão e capacidade
de associação com outros metais, permite o seu uso em soldas, fusı́veis, ligas metálicas e na
tipografia tradicional. É utilizado na indústria de tintas, vidros e cerâmicas na composição de
diversos pigmentos, sendo os mais comuns os brancos, na forma de carbonatos e sulfatos de
chumbo, e o amarelo, na forma de cromato de chumbo. Além disso, por ser um metal tóxico,
também é muito empregado como moluscicida na pintura de cascos de navios, como bactericida,
em especial para bactérias gram-positivas, e como fungicida. Desta forma, percebe-se que o
chumbo é um metal muito presente em nosso dia-a-dia, apesar de sua severa toxicidade[2, 3].
1.1 O chumbo como uma questão de saúde pública

Cientes dos efeitos tóxicos e nocivos do chumbo sobre o organismo, autoridades ao redor
do mundo empenharam-se nas últimas décadas para a redução a nı́veis aceitáveis deste tipo de
Considerações sobre o chumbo 14
contaminação ambiental. A gestão de resı́duos contaminados e a proibição do uso de aditivos à

base do metal nos combustı́veis são exemplos de medidas adotadas. Como consequência, o uso
indiscriminado de chumbo foi reduzido significativamente nos paı́ses industrializados. Entretanto,
alguns paı́ses em desenvolvimento possuem, ainda hoje, legislação bastante permissiva ou
inexistente no tocante ao uso do elemento e controle de seus rejeitos. É comum a utilização de
chumbo em diversos produtos oriundos de tais paı́ses, especialmente como compostos utilizados
em plásticos, tintas, equipamentos eletrônicos e baterias.
Desta forma, ainda existe uma ampla gama de fontes de exposição ao chumbo, onde se
pode citar:
• Poeira de tinta de casas antigas; • Fundições;
• Água proveniente de encanamentos anti-

• Empresas de reciclagem de baterias de
gos;
automóveis;
• Cerâmicas de baixa temperatura;
• Cosméticos;
• Joias;
• Medicamentos;
• Brinquedos;
• Rejeitos de mineradoras; • Alimentos contaminados; etc.
Alguns grupos populacionais também são especialmente suscetı́veis à contaminação por

chumbo, devido às suas atividades, padrões comportamentais ou caracterı́sticas biológicas, onde
se destacam as crianças, grupos ocupacionais, cujo trabalho os aproxima de fontes de chumbo
e populações desfavorecidas economicamente, cuja desnutrição pode acelerar a absorção do
metal [4].
Dentre todos os grupos de risco, o que mais causa preocupação é o das crianças, uma
vez que as alterações neurológicas causadas durante o desenvolvimento do sistema nervoso
central (do inglês Central Nervous System – CNS) podem ser consideradas irreversı́veis [5].
Organizações mundiais como as Nações Unidas e a World Health Organization, encontram-se
empenhadas na completa eliminação do uso de chumbo como pigmento de tintas, considerado
hoje uma das principais causas da intoxicação por chumbo em crianças [6]. Apesar destes
esforços, dados recentes ainda mostram nı́veis elevados de contaminação confirmada em
crianças com idade inferior a 72 meses nos Estados Unidos, paı́s que pode ser considerado
como padrão na regulamentação do uso do metal (Figura 1).
Uma vez que não há nenhuma função conhecida desempenhada pelo chumbo na fisiologia
humana, nenhum nı́vel de contaminação pode ser considerado seguro. O nı́vel máximo para
“desencadeamento de ações de combate à intoxicação” é de 5 µg/dL de sangue [8].
No Brasil, apesar de existir legislação sobre o uso e manuseio de rejeitos de produtos
contaminados por chumbo, esta encontra-se fragmentada dentro de diversas portarias e
resoluções de órgãos públicos, que também tratam de contaminação por diversos outros metais
Figura 1 – Número de casos confirmados de intoxicação por chumbo em crianças com

idade inferior a 72 meses ocorridos nos Estados Unidos entre os anos de 2012 e
2017. As linhas coloridas indicam os diferentes nı́veis de contaminação
medidos em µg/dL de sangue.
Fonte: US Center of Disease Control (CDC) [7]
e substâncias tóxicas, não existindo nem mesmo um nı́vel máximo de contaminação por chumbo
que seja oficialmente aceito [9, 10, 11]. Também não há dados oficiais sobre intoxicação
por chumbo. As ações de combate à contaminação são tomadas sob demanda e os casos
reportados em estudos são, em sua maioria, decorrentes de acidentes ambientais ou da iniciativa
de instituições de pesquisa [12, 13, 14].
1.2 Efeitos neurotóxicos do chumbo

O chumbo é uma neurotoxina de efeito acumulativo e excreção lenta, capaz de causar
danos irreversı́veis a diversos sistemas biológicos, depositando-se em tecidos como ossos, dentes
e sangue, com efeitos especialmente danosos ao sistema nervoso [5]. Quando armazenado em
ossos e dentes, pode ali permanecer por décadas, sendo paulatinamente liberado na corrente
sanguı́nea anos após os episódios de intoxicação. Adota-se internacionalmente o valor de
5µg/dL de sangue total como nı́vel máximo aceitável para intoxicação por chumbo.
Uma vez inalado ou ingerido, ele é rapidamente transportado à corrente sanguı́nea onde
irá ligar-se à membrana e proteı́nas intracelulares de eritrócitos. Apenas cerca de 3% de todo
chumbo absorvido permanecerá diluı́do no plasma sanguı́neo, permanecendo à disposição para
ser absorvido pelos demais tecidos do organismo [15, 16].
Este metal pesado pode acumular-se dentro das células intoxicadas, tanto de forma
aguda como crônica, associando-se a proteı́nas celulares e formando estruturas intracelulares
denominadas corpos de inclusão (do inglês inclusion bodies). Tais estruturas podem ser
particularmente observadas nos tecidos renal e cerebral [17, 18, 19, 20]. Afetando diversos
tecidos, a intoxicação por chumbo possui uma ampla gama de sintomas já bem descritos. Um
dos primeiros sintomas a aparecer em pessoas envenenadas por chumbo são as cólicas, náuseas
e vômitos. No sistema renal, pode causar nefrite tubulointersticial. No sistema cardiovascular,
tem sido associado a lesões cardı́acas, anormalidades eletrocardiográficas e aumento na pressão
sanguı́nea. No sistema musculoesquelético prejudica o desenvolvimento de ossos e dentes.
Estudos associam a esterilidade masculina a desordens no eixo HPT (Hipotálamo, Pituitária,
Testı́culos) causando, de forma ainda pouco conhecida, a redução da espermiogênese [21].
Também atribui-se como provável causa da esterilidade feminina a natural afinidade que o
chumbo possui com a gonadotrofina coriônica (hCG ), ligando-se diretamente a ela, quando irá
causar sua alteração conformacional e consequente inativação [22]. Outro mecanismo bastante
conhecido dá-se pela substituição do ı́on Zn2+ por P b2+ na proteı́na ALAD, impossibilitando
a sı́ntese Heme e, consequentemente, da hemoglobina (Hb), causando anemia no paciente
intoxicado [23].
Os efeitos da intoxicação por P b2+ são particularmente danosos quando tal exposição
ocorre durante o desenvolvimento do CNS, já que nestas condições tal sistema apresenta
uma maior capacidade de absorção de nutrientes e micronutrientes. Associado a desordens
cognitivas graves em crianças, são comuns os casos de pacientes intoxicados que apresentam
dificuldade de aprendizado e de consolidação da memória [23, 24, 25]. Mesmo quando em
baixas concentrações, problemas como alterações comportamentais, diminuição da capacidade
intelectual e de concentração foram reportados[26].
1.2.1 Causas da severa toxicidade do chumbo

Atribui-se a severa toxicidade do chumbo a sua capacidade de mimetizar ı́ons de Zn2+ e
Ca2+ . Estes dois metais participam de centenas de processos biológicos fundamentais à vida.
Desta forma, a sua substituição por chumbo acaba por causar um efeito tóxico generalizado
e sistêmico, afetando inúmeros componentes celulares com os quais tais ı́ons interagem e
interferindo em todas cascatas de sinalização por eles reguladas.
O papel do Zn2+
O zinco atua de forma direta ou indireta em inúmeros sistemas celulares, que envolvem
desde divisão celular, sı́ntese de DNA e sı́ntese proteica até a regulação do sistema imunológico.
É associado a muitas patogêneses neuronais, tais como a doença de Alzheimer, doença de
Parkinson, esclerose lateral amiotrófica, depressão e esquizofrenia, dentre outras [27, 28, 29].
Possui papel importante na sinalização de Ca2+ , uma vez que atua como bloqueador dos canais
de cálcio dependentes de voltagem [30]. Encontrado de forma abundante em tecido nervoso,
pode ser vinculado a terminais pré-sinápticos, em especial em neurônios glutamatérgicos, onde
regula a excitabilidade neuronal e a plasticidade sináptica. Também é associado a processos de
diferenciação neuronal e diferenciação morfológica de neurônios [27, 28, 29, 31].
Desempenha papel fundamental na estabilização da estrutura terciária de uma famı́lia de

proteı́nas denominadas zinc-fingers, compostas por domı́nios compactos contendo α hélices
e folhas β, cujas dobras e conformação são mantidas pelo Zn2+ . Codificadas por cerca de
3% do genoma humano, estas proteı́nas participam de inúmeros processos metabólicos, como
reconhecimento de DNA, empacotamento de RNA, ativação da transcrição, montagem e
folding de proteı́nas, regulação de apoptose, ligação de lipı́dios, etc. [23].
O papel do Ca2+
Assim como o zinco, o cálcio participa direta ou indiretamente de quase todos os processos
metabólicos humanos, atuando como fator de regulação na atuação de diversas enzimas e
proteı́nas.
Uma das principais proteı́nas afetadas pelo cálcio é a calmodulina (CaM), responsável pela
modulação de mais de 40 outras proteı́nas. Possuindo quatro sı́tios de ligação ao Ca2+ , tem
sua conformação alterada em função da abundância momentânea daquele ı́on, permitindo
sua interação com grupos diferentes de proteı́nas[32]. Abundante no CNS de mamı́feros, é
geralmente encontrada em locais envolvidos com a neurotransmissão, como nas membranas
pós-sinápticas, próximo à proteı́nas de densidade pós-sináptica (do inglês postsynaptic density
– PSD) e a vesı́culas sinápticas [33]. Dentre suas diferentes funções, pode-se destacar sua
participação no metabolismo de nucleotı́deos cı́clicos, na regulação de bombas Ca2+ , transporte
de ı́ons, fosforilação e desfosforilação de proteı́nas, montagem e desmontagem de microtúbulos,
sı́ntese de neurotransmissores, em especial a noradrenalina e serotonina, além da liberação de
neurotransmissores.
Além da CaM, outras proteı́nas dependentes de cálcio podem ser listadas como importantes
para o perfeito funcionamento do CNS. A proteı́na Quinase C (PKC) pode ser associada a
funções de controle do citoesqueleto de actina, de dinâmica de crescimento de microtúbulos, no
controle da plasticidade sináptica, onde regula os nı́veis de receptores NMDAR pós-sinápticos,
podendo ser assim correlacionada de forma direta ou indireta a diversas funções de aprendizado
e cognitivas [34]. As CaM-kinases (CaMK) têm papel fundamental no desenvolvimento e
funcionamento do neurônio, seja pela sua função na modulação de microtúbulos, seja na sua
atuação como fator ativo na potencialização sináptica durante o aprendizado [35]. Por fim,
o AMP cı́clico (do inglês Cyclic AMP – cAMP) pode também ser citado, já que participa
ativamente das cascatas de sinalização que implicarão nos processos de crescimento axonal,
interação entre astrócitos e neurônios, e na liberação de neurotransmissores [33, 23]. Um
resumo das principais vias metabólicas envolvidas e suas respectivas funções relacionadas ao
CNS podem ser vistas na Figura 2.
Figura 2 – Resumo de algumas vias metabólicas afetadas pelo cálcio e principais funções
do sistema nervoso central envolvidas.
1.3 O impacto sistêmico do envenenamento por chumbo

Uma vez introduzido no organismo, o chumbo irá causar impacto em diversos sistemas
metabólicos e em diferentes nı́veis dentro de cada um destes sistemas, perturbando ou im-
pedindo o funcionamento de muitas proteı́nas e enzimas, bem como alterando todo equilı́brio
homeostático iônico, podendo interferir nos seguintes processos, dentre outros:
• Diminuir a frequência de abertura de canais associados a receptores de acetilcolina;
• Interferir em canais de cálcio dependentes de voltagem, impedindo o seu correto fun-

cionamento e dificultando a entrada de cálcio no neurônio;
• Substituir ı́ons Ca2+ em metaloproteı́nas e receptores ionotrópicos, causando sua perda

de função, impedindo a transdução cálcio-dependente e interrompendo cascatas de
sinalização;
• Substituir o zinco como ligante nos zinc-fingers, alterando sua estrutura e interferindo
nos processos celulares por eles regulados;
• Substitui o Zn2+ em receptores NMDAR em vias glutamatérgicas, conhecidos por serem

fortemente modulados por este ı́on, causando inibição destes receptores e impedindo a
entrada dos ı́ons Ca2+ ;
• Causar estresse oxidativo, etc. [23]
Como consequência, uma diversa gama de sistemas metabólicos acabam por ser compro-
metidos, afetando processos neuronais que envolvem desde a expressão gênica até a liberação
de neurotransmissores, com consequências diretas sobre a capacidade de atenção, memória e
aprendizado.
Desta forma, o chumbo acaba por interferir, direta ou indiretamente, em inúmeros processos
celulares e afeta de forma sistêmica e global processos fundamentais ao bom funcionamento
das células. Um modelo esquemático das possı́veis interações do chumbo com os diversos
sistemas celulares pode ser visto na Figura 3.
Despolarização de Membrana Neurotransmissores e hormônios
Canais de Cálcio Adenilato Ciclase
OO Glutationa
O Glutationa Proteina Quinase C

Reductase
Glutationa
io
Peroxidase alc Transdução Cálcio
Estresse Oxidativo dec
ns Dependente
Glutataiona a io
Sintetase etiz
Mim
Pb2+ Receptores
ionotrópicos
Metaloproteínas Perda de
Função
Mimetiza ions de calcio
Aminolevulinato Ft Transcrição
Desidratase Zinc Fingers
5-Nucleotidase
EGR-1
Diferenciação Celular
SP1
Calmodulina
Sintese Heme Proteção e Concensação
Protamina P2
da Cromatina
Proliferação Apurinic/Apyrimidinic
Anemia
Celular Endonuclease 1
Reparo de DNA
Estresse Oxidativo
Legenda:
Estimulação
Mitocondria Inibição
Ligação Núcleo
Figura 3 – Modelo esquemático dos efeitos induzidos na célula pela intoxicação por
chumbo (traduzido de [23]).
20
2 Diferenciação celular
O processo de diferenciação celular ocorre em diversas fases sucessivas, onde células

não especialistas diferenciam-se em células cada vez mais especializadas, que irão formar os
diferentes tecidos encontrados em organismos complexos. O primeiro tipo de células nesta
cadeia de diferenciação são as células-tronco totipotentes, as quais podem se diferenciar
em qualquer outro tipo de células, incluindo-se aı́ células de tecidos extraembrionários. São
normalmente encontradas em tecidos embrionários até a fase de mórula e irão diferenciar-se
em duas linhagens: trofectoderma e as células-tronco pluripotentes. Estas últimas possuem o
potencial para diferenciarem-se em qualquer célula fetal ou adulta.
Quando isoladas, podem ser adaptadas e propagadas in vitro neste estado não diferenciado,
recebendo o nome de células tronco embrionárias (do inglês embryonic stem cells - ESC).
Através de protocolos adequados, tais células podem ser diferenciadas em células da endoderme,
mesoderme ou ectoderme, para, a seguir, tornarem-se células progenitoras de tecidos especı́ficos,
passando a ser classificadas como células multipotentes, capazes de se diferenciar em um
número limitado de células especı́ficas.
As células-tronco neurais são um exemplo deste tipo de células, sendo capazes de dar
origem a muitos, se não todos, os tipos de células gliais e neuronais do CNS [36]. Caso um
conjunto especı́fico de genes seja ativado neste momento, tais células sofrem nova diferenciação,
tornando-se células progenitoras neurais (NPCs), perdendo a capacidade de diferenciarem-se
em células da glia, podendo apenas se transformarem nos diversos tipos de neurônios. A Figura
4 ilustra todo este processo. As NPCs podem ser caracterizadas com base em sua localização
no cérebro, morfologia, distribuição temporal e perfil de expressão gênica.
Diferenciação celular 21
Figura 4 – Modelo esquemático da diferenciação neuronal a partir de células-tronco

pluripotentes. Uma célula pluripotente se especializa, diferenciando-se em uma
célula-tronco neural multipotente, que possui a capacidade de diferenciar-se
em células progenitoras gliais e neurais. Aquelas poderão se diferenciar em
diversos tipos de células que irão compor a glia e estas últimas poderão
diferenciar-se nos diversos tipos de neurônios. Os nomes em vermelho sobre a
célula progenitora neural e sobre o neurônio indicam alguns dos genes que são
caracteristicamente expressos nestes respectivos estágios da diferenciação
celular.
2.1 Caracterizando a diferenciação neuronal em um transcriptoma

A melhor forma de caracterizar a diferenciação de NPCs em neurônios a partir de dados tran-
scricionais é pela identificação do perfil de expressão de genes especı́ficos e caracterı́sticos destas
diversas fases, que atuarão como marcadores de tipo de especialização [37, 38]. Marcadores,
no caso em questão, podem ser definidos como sendo genes que, devido às suas caracterı́sticas
de expressão em fases especı́ficas do desenvolvimento celular, podem ser utilizados para realizar
a análise fenotı́pica do estágio de diferenciação celular. Assim, no presente trabalho, foram
empregados dois tipos distintos de marcadores: marcadores para NPCs e marcadores neuronais.
2.1.1 Genes marcadores de NPCs

Musashi1 (MSI1)
O MSI1 é um gene cujos valores de expressão se apresentam enriquecidos nas fases iniciais
do desenvolvimento do CNS. Sua elevada taxa de transcrição ocorre durante o desenvolvimento
cerebral, principalmente em NPCs, células da camada granular externa cerebelar, ependimócitos
e astrócitos, mas nunca em neurônios ou oligodendrócitos. Podem ainda ser encontrados

traços de sua expressão em neurônios jovens, mas que vão desaparecendo conforme ocorre sua
maturação [39].
Nestina (NES)
Inicialmente identificada em células tronco neuroepiteliais de ratos, a Nestina é um indicador

importante da neuroplasticidade das células. A expressão dos genes produtores de Nestina é
encontrada em elevados nı́veis em células da glia radial, neurolemócitos (células de Schwann),
células da crista neural, precursores de oligodendrócitos, células gliais de retina (células de
Müller) e NPCs, dentre outras células em diferenciação [40].
NOTCH1
Notch1 é um gene que pode ser correlacionado à diferenciação celular em diversos tecidos
cerebrais. Sua deficiência durante os estágios iniciais de desenvolvimento do CNS pode levar à
falha de diferenciação e à apoptose celular, especialmente em células da glia. Existem indı́cios
que sugerem que muitos mecanismos de diferenciação de células tronco neurais, incluindo-se aı́
as NPCs, são controlados pelas rotas de sinalização do Notch1 [41].
SOX1
Sox1 é um membro da famı́lia de fatores de transcrição SoxB1 e sua expressão está

correlacionada aos passos iniciais da neurogênese. A expressão forçada deste gene, provocada
in vitro, mostra expressivo aumento na taxa de geração de células da neuroectoderme e de
neurônios [42].
2.1.2 Genes marcadores neuronais

Tirosina Hidroxilase (TH)
A TH é uma enzima considerada crı́tica para o funcionamento de diversos tipos de neurônios,

como neurônios adrenérgicos, noradrenérgicos e, principalmente, neurônios dopaminérgicos.
Essencial na conversão da L-Tyrosina em L-DOPA, onde atua como catalizador, é elemento
indispensável na cascata que produzirá a dopamina, noradrenalina e adrenalina, sendo assim
elemento fundamental na caracterização de atividade neuronal. Baixos nı́veis de produção de
TH estão relacionadas a doenças do CNS, especialmente o mal de Parkinson [43].
NEUROD6
Membro da subfamı́lia NEUROD, pertencente ao grupo dos fatores de transcrição basic-

helix-loop (bHLH), o NEUROD6 é indispensável ao crescimento dos axônios do neocortex.
Seus elevados nı́veis de expressão estão associados ao desenvolvimento de neurônios maduros a

partir de neurônios imaturos, em especial os dopaminérgicos [44].
Doublecortina (DCX)
A doublecortina (DCX) é uma fosfoproteı́na necessária ao desenvolvimento dos microtúbulos.

É associada à diferenciação neuronal, sendo expressa por ocasião da migração de neurônios
no sistema nervoso central durante o seu desenvolvimento. É considerada um marcador de
neurogênese, particularmente relacionada à geração de novos neurônios no gyrus dentatus do
hipocampo [45, 46].
RBFOX3
Codificada pelo gene RBFOX3, a Proteina Neuronal Nuclear (NeuN) é comumente encon-
trada nos núcleos e citoplasma perinuclear da maioria dos neurônios do CNS de mamı́feros.
É frequentemente utilizada como um marcador neuronal devido a esta sua propriedade lo-
calizacional. Anticorpos monoclonais à proteı́na NeuN têm sido utilizados ativamente na
determinação da existência de diferenciação neuronal e na avaliação do estado funcional de
neurônios [47].
GAD1 e GAD2
Estes genes são responsáveis pela codificação das formas 1 e 2 da descarboxilase do ácido
glutâmico. Estas enzimas catalizam a produção de ácido gama-aminobutı́rico a partir do ácido L-
glutâmico. Também conhecidos como GAD67 e GAD65, respectivamente, estes genes, além de
desempenhar funções no pâncreas, relacionando-os ao diabetes do tipo 1 (insulinodependente),
são considerado marcadores de diferenciação celular de neurônios GABAérgicos, estando sua
expressão intimamente correlacionada com os nı́veis de expressão da TH neste tipo de neurônio
[48].
24
3 Análise do transcriptograma
3.1 O pacote transcriptogramer

Para realizar uma análise sistêmica do transcriptoma foi utilizado o pacote transcriptogramer
do R, desenvolvido pelo nosso grupo de trabalho. Ele presta-se a realizar uma análise de
transcriptomas baseada em biologia de sistemas, que, em última análise, permite a identificação
de grupos funcionais de genes diferencialmente expressos em experimentos do tipo caso-
controle, pois provê ferramentas para criação de transcriptogramas, análise topológica dos
dados, análise de expressão diferencial e de enriquecimento de ontologia de genes (GO) [49, 50].
O método utiliza uma rede de interação proteı́na-proteı́na (PPI) extraı́da do STRINGdb,
ordenada unidimensionalmente de acordo com a probabilidade de interação entre os diferentes
pares de proteı́nas.
3.1.1 Sequencia ordenada de genes

Conforme o método descrito em [50], de posse da rede PPI, cria-se uma matriz simétrica de
interação proteı́na-proteı́na de N × N elementos, onde posições contendo o valor 1 representam
a interação entre duas proteı́nas quaisquer e 0 a inexistência de interação, como pode ser visto
no exemplo da Figura 5(a). Calcula-se, então, o custo E desta matriz aleatória utilizando-se
a equação 3.1, onde dij representa a distância dos diferentes pares de pontos da matriz em
relação à diagonal principal. Valores elevados de dij representam proteı́nas que interagem, mas
encontram-se em posições distantes na matriz. Assim, o desejado é reorganizar as proteı́nas de
forma que esta nova ordem possua o menor custo, garantindo que proteı́nas com algum tipo
de interação encontrem-se o mais próximo possı́vel. Para tanto, o algoritmo Cost Function
Minimizing (CFM) toma duas proteı́nas aleatoriamente, troca-as de posição ( Figura 5(b)
) e calcula o novo custo E 0 . Caso este seja menor que o valor inicial, a mudança é aceita.
Caso contrário ela é rejeitada. Nova troca aleatória é realizada até que o valor de E tenha se
estabilizado (Figura 5(c)).
N X
X N
E= dij {|Mi,j − Mi+1,j | + |Mi,j − Mi−1,j | + |Mi,j − Mi,j+1 | + |Mi,j − Mi,j−1 |}
i j
(3.1)
A Figura 6 representa uma matriz PPI de S. cerevisiae [50] antes e após o ordenamento.
Os clusteres observados próximos à diagonal principal assinalam regiões onde existem grande
interação entre proteı́nas vizinhas. Ao término deste procedimento, obtém-se uma lista de
genes/proteı́nas, ordenadas pela similaridade de processos biológicos dos quais participam. Tal
ordenamento será utilizado pelo transcriptogramer como eixo dos X.
Análise do transcriptograma 25
(a) Ordem aleatória (b) Troca aleatória de posição (c) Ordenado
Figura 5 – Matriz PPI de exemplo. As letras representam proteı́nas e os números 1

representam as interações entre elas. Proteı́nas que não interagem entre si
recebem o valor 0 na célula correspondente da matriz de adjacência (omitido
na figura para fins de clareza). (a) Estado inicial da matriz em uma ordem
aleatória qualquer. Neste estado é calculado o custo inicial E da matriz,
segundo a equação 3.1. (b) As proteı́nas B e J são aleatoriamente escolhidas e
tem suas posições trocadas, sendo o novo custo E 0 calculado. (c) Estado final
do processo, onde E se estabiliza em um valor mı́nimo.
(a) Matriz PPI em ordem aleatória (b) Matriz PPI ordenada
Figura 6 – (a) Exemplo de matriz PPI em uma ordem aleatória. (b) A mesma matriz
após a aplicação do algoritmo de ordenação Cost Function Minimizing (CFM)
(figura adaptada de [50])
3.1.2 Expressão diferencial (DE)

O pacote transcriptogramer realiza o cáculo da expressão diferencial utilizando-se das
funcionalidades do pacote limma (Linear Models for Microarray Data [51]). Este último faz
uso de modelos lineares a fim de avaliar a expressão diferencial em experimentos de RNA-seq
ou microarray, do tipo caso-controle com múltiplas amostras. Em suma, o transcriptogramer

fornece ao pacote limma os valores de expressão das diversas amostras disponı́veis e um vetor
lógico, onde tais amostras são identificadas como sendo caso ou controle, e obtém como
resultado valores de DE dos diversos genes/proteı́nas e um p-valor associado.
3.1.3 Valores médios de expressão – Janela deslizante

De posse da lista ordenada de proteı́nas e de seus valores de expressão diferencial (microarray
ou RNA-seq) entre caso-controle, aplica-se uma janela deslizante de raio R arbitrário, conforme
ilustrado na Figura 7(a). Os valores de expressão diferencial encontrados sob a janela tem a
sua média calculada e esta passa a representar o valor de DE da posição central da janela.
Assim, tal valor pode ser interpretado como a DE de uma região do interatoma que envolve
um número de proteı́nas contı́guas igual a 2R + 1, já que proteı́nas encontradas em posições
vizinhas tem afinidade funcional, garantida pelo ordenamento prévio. A Figura 7(b) ilustra
a situação final do processo envolvendo a janela deslizante e a Figura 8, sua representação
gráfica.
Apesar de não haver um tamanho determinado para o raio R, o uso de valores entre 70 e
120 tem se mostrado adequado.
(a) Lista ordenada de proteı́nas e seus respectivos valores de expressão
(b) Resultado final após a aplicação da janela deslizante
Figura 7 – Exemplo de lista ordenada de proteı́nas, com respectivos valores de expressão

diferencial (DE), onde será aplicada uma janela deslizante de raio igual a 2.
(a) Os valores sob a janela tem sua média calculada e esta passa a ser o valor
de referência de DE da posição central da janela. A janela é então deslocada
para a direita e o processo se repete. (b) Situação final após a janela percorrer
todas as posições da lista ordenada.
Figura 8 – Exemplo de representação gráfica das médias de DE após o processo de janela

deslizante. O eixo dos X representa as posições da lista ordenada de proteı́nas.
O eixo dos Y representa os valores de DE dentro das diferentes janelas. A
linha horizontal preta representa os valores de expressão de referência do
controle e a linha cinza os valores médios de expressão do caso.
3.1.4 Clusterização
De forma semelhante ao processo utilizado na obtenção dos valores médios de DE por janela
deslizante, os clusteres funcionais considerados significativos são obtidos pela utilização de uma
janela de mesmo raio que irá deslizar por sobre os p-valores obtidos do pacote limma. Um
cluster irá crescer sempre que dentro da janela houver pelo menos dois p-valores menores que
um limiar predeterminado, sendo os limites de tal cluster estendido até a posição do segundo
p-valor considerado significativo. A Figura 9 ilustra o processo onde uma janela deslizante de
raio 2 encontra dois p-valores menores que 0,01 nas posições 5 e 7 do ordenamento, estendendo
o cluster 1 até aquela última posição. A Figura 10 ilustra a representação gráfica dos clusteres
por sobre o gráfico de médias de DE.
Figura 9 – Exemplo de lista ordenada de proteı́nas, com respectivos p-valores de DE,

onde será aplicada uma janela deslizante de raio igual a 2 para determinação
de clusteres significativos.
Figura 10 – Exemplo de representação gráfica das médias de DE e dos clusteres funcionais

considerados significativos, identificados por cores.
3.1.5 Enriquecimento dos clusteres por gene ontology (GO)

O enriquecimento de GOs presentes dentro de cada cluster é realizado tomando-se a relação
de genes/proteı́nas que participam de cada um dos clusteres identificados, apresentando-as ao
pacote topGO [52]. Como resultado, obtém-se uma lista de ontologias que foram consideradas
relevantes. A Tabela 1 ilustra um exemplo de resultado de algumas GOs consideradas relevantes
em um hipotético cluster 3, onde podem ser identificados os ID das GOs, a descrição de cada
termo e a significância do enriquecimento, representada pelo p-valor.
Cluster GO.ID Term Annotated Significant Expected pValue

GO:0006811 ion transport 1299 46 17.73 2.93e-05
3 GO:0051259 protein complex oligomerization 445 17 6.07 1.83e-05
GO:0055085 transmembrane transport 1229 42 16.77 3.32e-04
1
Tabela 1 – Exemplo de enriquecimento de ontologias.
3.1.6 Rede hierárquica de termos de GO

A fim de facilitar a interpretação das GOs enriquecidas, desenvolvemos uma ferramenta
que constrói um dendrograma contendo a hierarquia das ontologias de um determinado cluster.
Para tanto, ele calcula o ı́ndice Jaccard de cada par de GOs criando uma matriz de adjacência
utilizada na construção daquele dendrograma. Um exemplo desta rede hierárquica pode ser visto
na Figura 11. Esta função estará disponı́vel em versões futuras do pacote transcriptogramer.
Figura 11 – Exemplo de uma rede hierárquica de termos de GO. Os tamanhos dos nós são
proporcionais ao números de genes associados à GO. As cores dos nós
representam a razão normalizada entre o número de genes enriquecidos
presentes na GO pelo número total de genes associados a ela.
1
A coluna Cluster informa o identificador do cluster ao qual as GOs pertencem. A coluna GO.ID
mostra o identificador da GO que apresentou enriquecimento. A coluna Term exibe a descrição da GO.
A coluna Annotated informa o total de genes estão anotados na GO. A coluna Significant informa
a quantidade de genes pertencentes ao cluster que foram identificados na GO. A coluna Expected
informa a quantidade máxima de genes que seria esperada em uma distribuição aleatória. E a coluna
pValue informa o p-valor corrigido, obtido no teste estatı́stico de enriquecimento.
30
Justificativa
O chumbo é um insumo praticamente indispensável à vida moderna. Seus efeitos nocivos

graves, cujos sintomas já são bem conhecidos, causam sequelas muitas vezes irreversı́veis à
saúde humana, em especial ao sistema nervoso central, onde a exposição precoce em crianças
leva a disfunções neuronais permanentes com consequente defasagem no desenvolvimento
intelectual, aprendizagem e processos envolvendo memória. Ao longo de anos, diversos estudos
toxicológicos na área foram levados a cabo e foram identificadas uma série de proteı́nas capazes
de interagir com o chumbo, cujas associações explicam alguns dos muitos sintomas observados.
Porém as análises de tais interações ocorrem, via de regra, de forma estanque e isolada.
Apesar dos esforços e avanços, nota-se que tais interações carecem de um olhar mais
abrangente, onde a funcionalidade de diversos componentes celulares possa ser observada como
um conjunto integrado. Assim, uma análise sistêmica dos efeitos do chumbo sobre a atividade
neuronal faz-se necessária para um melhor entendimento da influência deste metal pesado nas
diversas vias metabólicas e redes de interação, de forma que se possa ter uma compreensão
holı́stica dos processos envolvidos e da sintomatologia decorrente.
31
Objetivos
Objetivo geral
– Analisar o perfil transcricional de células progenitoras neurais humanas (NPCs) tratadas
com acetato de chumbo, de forma a identificar, sob uma ótica sistêmica, as possı́veis
consequências da exposição ao chumbo de células neurais em desenvolvimento.
Objetivos especı́ficos
– Identificar marcadores que permitam caracterizar a diferenciação de células neuronais
nas amostras selecionadas;
– Identificar, em um esquema caso/controle, grupos diferencialmente expressos, clusterizando-

os por similaridade funcional, pela utilização do pacote R/Bioconductor transcriptogramer ;
– Caracterizar as redes de interação proteı́na-proteı́na moduladas, baseadas nas anotações

do Gene Ontology ;
– Analisar os padrões de expressão diferencial encontrados, correlacionando-os com al-

terações metabólicas relacionadas ao envenenamento por chumbo descritas na literatura.
Artigo:
Systems Biology-Based Analysis Indicates
Global Transcriptional Impairment in
Lead-Treated Human Neural Progenitor Cells
Autores: Clovis F. Reis, Iara Souza, Diego Arthur D. Morais, Raffael A. Oliveira, Danilo O.
Imparato, Rita M. De Almeida e Rodrigo J. Dalmolin
Artigo publicado em 10 de Setembro de 2019

Frontiers in Genetics
doi: 10.3389/fgene.2019.00791
ORIGINAL RESEARCH
published: 10 September 2019
doi: 10.3389/fgene.2019.00791
Systems Biology-Based Analysis

Indicates Global Transcriptional
Impairment in Lead-Treated Human
Neural Progenitor Cells
Clovis F. Reis 1†, Iara D. de Souza 1†, Diego A. A. Morais 1, Raffael A. C. Oliveira 1,
Danilo O. Imparato 1, Rita M. C. de Almeida 2 and Rodrigo J. S. Dalmolin 1,3*
1Bioinformatics Multidisciplinary Environment — IMD, Federal University of Rio Grande do Norte, Natal, Brazil, 2 Institute of
Physics and National Institute of Science and Technology: Complex Systems, Federal University of Rio Grande do Sul, Porto
Alegre, Brazil, 3 Department of Biochemistry — CB, Federal University of Rio Grande do Norte, Natal, Brazil
Lead poisoning effects are wide and include nervous system impairment, peculiarly
Edited by: during development, leading to neural damage. Lead interaction with calcium and zinc-
Argyris Papantonis,
University Medical Center
containing metalloproteins broadly affects cellular metabolism since these proteins are
Göttingen, Germany related to intracellular ion balance, activation of signaling transduction cascades, and gene
Reviewed by: expression regulation. In spite of lead being recognized as a neurotoxin, there are gaps in
Chiara Piubelli, knowledge about the global effect of lead in modulating the transcription of entire cellular
Ospedale Sacro Cuore
Don Calabria, Italy systems in neural cells. In order to investigate the effects of lead poisoning in a systemic
Marco Vanoni, perspective, we applied the transcriptogram methodology in an RNA-seq dataset of
University of Milano-Bicocca,
human embryonic-derived neural progenitor cells (ES-NP cells) treated with 30 μM lead
Italy
acetate for 26 days. We observed early downregulation of several cellular systems involved
*Correspondence:
Rodrigo J. S. Dalmolin with cell differentiation, such as cytoskeleton organization, RNA, and protein biosynthesis.
rodrigo.dalmolin@imd.urfn.br The downregulated cellular systems presented big and tightly connected networks. For
† These authors have contributed long treatment times (12 to 26 days), it was possible to observe a massive impairment in
equally to this work cell transcription profile. Taking the enriched terms together, we observed interference in
all layers of gene expression regulation, from chromatin remodeling to vesicle transport.
Specialty section:
This article was submitted to Considering that ES-NP cells are progenitor cells that can originate other neural cell
Systems Biology, types, our results suggest that lead-induced gene expression disturbance might impair
a section of the journal
Frontiers in Genetics
cells’ ability to differentiate, therefore influencing ES-NP cells’ fate.
Received: 26 March 2019 Keywords: lead exposure, lead poisoning, transcriptogramer, RNA-seq, transcriptome analysis, network
Accepted: 26 July 2019 inference, data integration, network visualization
Published: 10 September 2019
Citation:
Reis CF, de Souza ID, Morais DAA, INTRODUCTION
Oliveira RAC, Imparato DO,
de Almeida RMC and Dalmolin RJS Lead is largely used in industry and is very toxic to biological systems. This heavy metal
(2019) Systems Biology-Based accumulates in hard tissues, remaining in bones and teeth for decades (Ronis et al., 2001).
Analysis Indicates Global
Lead systemic effects can be observed through a wide range of lead poisoning symptoms. It
Transcriptional Impairment in
Lead-Treated Human
includes anemia, abdominal pain, vomiting, cardiovascular system impairment, nephropathies,
Neural Progenitor Cells. and abnormal spermatogenesis (Flora et al., 2012; Mitra et al., 2017). Several studies describe
Front. Genet. 10:791. lead toxicity in central nervous system as well as its relation to irreversible brain development
doi: 10.3389/fgene.201900791. impairment (Finkelstein et al., 1998; Baranowska-Bosiacka et al., 2012; Stansfield et al., 2012).
Frontiers in Genetics | www.frontiersin.org 1 September 2019 | Volume 10 | Article 791

Reis et al. Transcriptogram Analysis of Lead-Treated NPCs
This diversity of systemic effects indicates that lead interacts MATERIALS AND METHODS
with many cellular components. A recent review conducted
by our group highlighted proteins that are described to Data Selection and Preprocessing
directly interact with lead and how those interactions could Data from 26 RNA-seq of human embryonic-derived neural
be related to lead poisoning symptoms (de Souza et al., 2018). progenitor cells (ES-NP cells) samples treated with lead acetate
Lead strongly inhibits NMDA receptors, a transmembrane 30 μM in a time-series experiment (day 1 to day 26) and 27
calcium channel from glutamatergic neuronal pathways that respective control samples (day 0 to day 26) were obtained from
performs a crucial role in brain development (Nihei et al., Gene Expression Omnibus (accession GSE84712) (Jiang et al.,
2000; Baranowska-Bosiacka et al., 2012). NMDA receptor 2017). Raw data sequence reads were downloaded from Sequence
inhibition is related to lead binding in the NMDA receptor Read Archive (accession SRP079342) and processed following
zinc-binding site. In fact, much of lead toxicity relies on its new Tuxedo protocol (Pertea et al., 2016) using Ensembl
binding to metalloproteins with divalent ions as prosthetic GRCh38 Human genome reference and annotation (release 91).
groups, especially calcium and zinc. Metalloproteins are Counts were then filtered, normalized, and converted into log2-
involved with a variety of cellular processes, from regulation counts-per-million (log-CPM) following limma R/Bioconductor
of cellular ion balance to activation of signaling cascades, package protocol (Ritchie et al., 2015). Principal component
inducing changes in cell metabolism. Calcium-interacting analysis (PCA) was performed with all samples in order to check
proteins perform important roles in cell transduction data quality (Supplementary Figure S1).
cascades. Lead mimics calcium on protein kinase C (PKC)
isoforms and calmodulin (CaM), triggering their downstream
signaling cascades (Richardt et al., 1986; Sun et al., 1999). Lead Group Selection and Transcriptogram
interacts with zinc finger transcription factor family members Analysis
by substituting zinc ions, impairing zinc fingers DNA binding, Differential expression of functionally associated gene groups
and directly interfering in gene expression (Zawia et al., 1998; among ES-NP cells treated with lead acetate 30 μM and
Reddy and Zawia, 2000; Ghering et al., 2005). Taken together, respective controls was performed using transcriptogramer
lead interaction to metalloproteins can massively interfere at R package (Morais et al., 2019). The transcriptogramer is a
the cell transcription profile (Guilarte and McGlothan, 1998; systems biology-based method to analyze transcriptomes,
Ordemann and Austin, 2016). which uses PPI information to identify differentially expressed
Here, we investigate lead poisoning transcriptional effects functionally associated gene groups in case-control designed
from a systemic perspective by applying the transcriptogram experiments (Rybarczyk-Filho et al., 2011; Morais et al., 2019).
pipeline to a lead poisoning experiment data publicly available. Briefly, the method sorts the genes in one dimension by the
Transcriptogram pipeline consists of a systems biology-based probability that their products collaborate in a biological
methodology designed to perform transcriptional analysis function and uses the resulting ordered gene list to project
of functionally associated gene groups in case-control the expression of functionally associated genes in a sliding
experiments (Rybarczyk-Filho et al., 2011). It implements a window with a given radius. As a result, the transcriptogramer
non-supervised approach based on protein–protein interaction evaluates the expression of entire genetic/biochemical systems
(PPI) to identify differentially expressed protein subnetworks. in a biological condition. Transcriptogramer package uses the
Transcriptogram pipeline results in global transcriptional limma algorithm to compute the significance of functionally
profiles of each biological phenomenon evaluated, evidencing associated gene groups expression variation in case-control
the biochemical systems collectively altered in consequence experiments, identifying differentially expressed clusters in a
of a given disturbance. We analyzed an RNA-seq experiment non-supervised way.
of human embryonic-derived neural progenitor cells (ES- To allow transcriptogramer statistical evaluation, lead-
NP cells) treated with 30 μM lead acetate for 1 to 26 days treated samples were grouped by expression similarity using
(Jiang et al., 2017). Our analysis has focused on identifying PCA clustering. Clustering analysis was conducted on the
early and late global lead-induced transcriptional alterations. 30 μM lead-treated samples using the first 17 principal
Supporting previous findings from Jiang et al. (2017), our components, which corresponds to about 95% of the
results showed that lead interferes in many cellular processes cumulative variance (Supplementary Figure S2). Group
(e.g., cell cycle regulation, macromolecule metabolism, selection was performed by a non-supervised algorithm using
response to DNA damage, and cytoskeleton organization). HCPC function from FactoMineR R/CRAN package (Lê et al.,
Additionally, our analysis pointed to different levels of 2008) and methodology from Husson et al. (2010). Clustering
lead-induced perturbation when considering early and late was carried out using automatic cluster detection, identifying
exposure. It was possible to observe a high transcriptional three groups of samples. Group 1, defined as time-interval 0,
downregulation of well-connected systems at initial days of included only two samples (day 1 and 2) and therefore was
exposure. In a second time interval, we observed an overall discarded for transcriptogramer analysis. The remaining two
transcriptional modification suggesting that lead exposure groups were defined for the study as time-interval 1 (days
results in massive interference in gene expression regulation 3 to 11; n = 9) and time-interval 2 (days 12 to 26; n = 15)
in ES-NP cells. (Supplementary Figure S3).

Progression of the Neuronal Markers (i.e., the number of genes associated to the GO term), and the
To characterize the differentiation of the neural progenitors node color represents the ratio of significantly enriched genes
(NPCs) to neuronal cells, we evaluated the expression (log-CPM) over the count of all annotated GO term genes, normalized by
of a set of well-known markers for NPCs and neurons. We used the max ratio found in the cluster. Clusters dendrograms were
the genes MSI1, NES, NOTCH1, and SOX1 as NPC markers, generated using 0.25 Jaccard as a cutoff parameter, and the
and genes TH, NEUROD6, DCX, RBFOX3, GAD1, and GAD2 final layout was shown using RedeR R/Bioconductor package
as neuronal markers (Kaneko et al., 2000; Lutolf et al., 2002; (Castro et al., 2012).
Wiese et al., 2004; Cossette et al., 2005; Suter et al., 2009; White
and Thomas, 2012; Gusel’nikova and Korzhevskiy, 2015; Khan
et al., 2017; Mathews et al., 2017). As explained in the section RESULTS
above, the transcriptional data used here are a time-series
experiment including 53 samples: 26 lead-treated (lead acetate
Expression Markers for ES-NP Cells
30 μM) samples (day 1 to day 26), 26 control samples (day 1 to Differentiation
day 26), and day 0 sample. To test marker expression variation ES-NP cells are neural progenitor cells able to differentiate into
by time, we subdivide both control and treated samples into neurons and glial cells (Selvaraj et al., 2012) and the modified
three groups each: i) time-interval 0 (from day 0 to 2), ii) time- protocol used by Jiang and collaborators points to neuronal
interval 1 (from day 3 to 11), and iii) time-interval 2 (from day differentiation (Chambers et al., 2009; Jiang et al., 2017). The
12 to 26). The same unique day 0 sample was used to compose samples comprise a time-course experiment from 26 RNA-seq
control time-interval 0 and treated time-interval 0. The marker of human embryonic-derived neural progenitor cells (ES-NP
expression differences among the time intervals (for both cells) samples treated with lead acetate 30 μM (day 1 to day 26)
control and lead-treated) were obtained by a pairwise t-test and 26 respective control samples (day 1 to day 26) plus day 0.
using the pairwise.t.test function from stats R package. The The treated samples were grouped by PCA followed by non-
p-values were corrected by the FDR method, and p-values below supervised clustering, producing three groups of samples:
0.01 are considered significant. i) time-interval 0, comprising days 0 to 2; ii) time-interval 1,
comprising days 3 to 11; and iii) time-interval 2, comprising days
12 to 26 (see Materials and Methods and Supplementary Material
Transcriptogramer Analysis Online for details). We evaluated the expression dynamics of
Transcriptogramer differential expression method was applied to neuroprogenitor cells (NPCs) marker genes, as well as neuronal
each time interval comparing treated groups with the respective cell marker genes, to evaluate the ES-NP cells differentiation
control, using ordered gene list built under STRING V10 progression (Figure 1, Supplementary Figures S4 and S5).
combined score ≥700, window radius = 80. Differential expression Figure 1 shows the expression of NPC markers (NES, NOTCH1,
statistical relevance was assessed by Fisher test with Benjamini- MSI1, and SOX1) and neuronal cell markers (DCX, GAD1,
Hochberg adjustment of p-values (p-value ≤ 0.001). After GAD2, NEUROD6, TH, and RBFOX3) in both control cells and
differential expression analysis, the PPI networks of significant treated cells. NES and MSI1 are described as highly expressed
clusters were plotted using clusterVisualization transcriptogramer in progenitor-like cells (Kaneko et al., 2000; Wiese et al., 2004).
function, and GO enrichment analysis of each differentially NOTCH1 and SOX1 are also described as highly expressed in
expressed cluster was performed using clusterEnrichment NPCs and are the main drivers of differentiation process (Lutolf
function. Final network manipulation was performed using et al., 2002; Suter et al., 2009), while DCX is an important
RCytoscape R/Bioconductor (Shannon et al., 2013) and RedeR marker for immature neurons (Mathews et al., 2017). TH and
R/Bioconductor packages (Castro et al., 2012). Connectivity NEUROD6 are recognized as dopaminergic markers (White and
graphs (Figures 2B, D) represent the average network Thomas, 2012; Khan et al., 2017). GAD1 and GAD2 are gabaergic
connectivity and were plotted using the circlize R/CRAN package differentiation markers and were found to have a good correlation
(Gu et al., 2014). with TH expression in gabaergic neurons (Cossette et al., 2005).
RBFOX3 is a postmitotic neuronal marker (Gusel’nikova and
Korzhevskiy, 2015). Figure 1 shows the mean expression levels
GO Terms Hierarchical Networks of the aforementioned genes in each time interval for control
The hierarchical networks of significantly enriched GO terms and treated cells. The majority of NPC markers decreased their
in each differentially expressed cluster were built based on expression in both control and lead-treated samples when
the similarity among any pair of enriched GO terms (i.e., comparing the initial and final time intervals. The exception is
the number of shared genes in each pair of GO terms). The NOTCH1 expression, which was significantly altered in control
method calculates the Jaccard’s Index of each GO term pair, samples only when comparing time-interval 1 and 2, but not when
all-against-all, and builds an adjacency matrix that is used comparing time-interval 0 and 2. Neuronal marker expression
to compute a dendrogram. The resulting dendrogram is then increased in the same period in both control and lead-treated
plotted as a tree-and-leaf network using RTN R/Bioconductor samples, except for TH expression, which was not significantly
package (Fletcher et al., 2013; Castro et al., 2016). Each node altered in lead-treated samples (Figure 1). The dynamics of
in the resulting hierarchical network represents a significantly NPC and neuronal marker expression observed in Figure 1 is
enriched GO term. Node size is proportional to GO term size consistent with neuronal differentiation since NPC marker

expression decreased while neuron marker expression increased contributes proportionally to the time-interval 1 interactome
along with the experiment. However, the markers transcriptional (Supplementary Figure S18). Collectively, downregulated
dynamics are significantly different when comparing treated and clusters are more connected when compared to upregulated
control samples (Supplementary Figures S4 and S5). clusters on time-interval 1 (Figure 2B).
Transcriptogramer analysis of time-interval 2 shows a global
alteration in cell transcription pattern, suggesting a progressive
Transcriptional Profile and Clusters cellular response among time intervals. As shown in Figure 2C,
Network Dynamics on Time Intervals many clusters merged as they became larger. The merged clusters
To evaluate the transcriptional disturbance caused by lead, we in time-interval 2, as well as clusters identified only on this
applied the transcriptogram pipeline to time-interval 1 and 2, time interval, were represented by letters (Figure 2C). At total,
always comparing lead-treated samples to their respective almost 90% of lead-treated ES-NP cell interactome was altered
control samples (see Materials and Methods and Supplementary in time-interval 2. All upregulated clusters in time-interval 1
Material Online for details). The transcriptogram is a systems expanded in time-interval 2: clusters 1, 2, and 3 merged into
biology-based method for transcriptional analysis, which uses cluster A and cluster 4 drastically increased in time-interval 2.
PPI information to build an ordered gene list where interacting Additionally, upregulated clusters increased their transcriptional
genes are expected to get closer to each other (Rybarczyk- differences against the control group (black line in Figure 2C).
Filho et al., 2011). The ordered gene list is then used to project Downregulated peaks decreased against the control group
the expression of functionally associated gene groups that are when compared to time-interval 1. It indicates a strong early
likely to participate in common biochemical pathways. modulation of downregulated clusters that decreases with time-
Transcriptogramer analysis of time-interval 1 identified interval transition. Cluster average connectivity is more equalized
11 clusters of differentially expressed gene sets, which were in time-interval 2 where the upregulated clusters enhanced their
sequentially numbered (Figure 2A). Clusters 1 to 4 were average connectivity (Figure 2D).
upregulated while clusters 5 to 11 were downregulated The dynamic involving the number of nodes and the average
(Figure 2A). Figure 2B represents the subnetwork involving connectivity of each cluster in the different time intervals can
the 11 differentially expressed clusters of time-interval 1. All be better observed in Figure 3. It is possible to observe that all
clusters have more inner than outer connections, indicating upregulated clusters in time-interval 1 strongly increased in
that methodology was able to identify functionally associated number of nodes in time-interval 2 (Figure 3, clusters A and 4).
gene groups. This representation illustrates how each cluster While cluster 4 highly increased the average connectivity,
clusters 1, 2, and 3 (merged as cluster A in time-interval 2)
average connectivity barely changed. Clusters identified as
downregulated in time-interval 1 also increased in time-
interval 2, except by clusters 6 and 8. However, the expansion
observed in downregulated clusters was smaller when compared
with the expansion experienced by upregulated clusters in time-
interval transition. In spite of increasing number of nodes, clusters
identified as downregulated in time-interval 1 showed a slight
decrease in average connectivity in time-interval 2, except by
clusters 5 and 11 that have a particular dynamic on time-interval
transition: they started as downregulated clusters on time-
interval 1 and expanded on time-interval 2, and then presented
down- and upregulated portions on the transcriptogram. The
expansion in the number of nodes observed in clusters 5 and 11
was followed by an increase in average connectivity.
FIGURE 1 | NPC and neuronal marker progression. Left graphics show the Functional Enrichment of Upregulated
control group average expression values in log-CPM, for NPC markers (on
top) and neuronal markers (on bottom). Right graphics show the lead-treated
Clusters in Time-Interval 1
sample average expression values. Colors represent distinct markers.
According to our data, lead treatment was able to impair
Numbers at the X-axis identify the time intervals starting on zero, for samples ES-NP cell global expression in both time intervals, either by
from day 0 to 2, and time-intervals 1 and 2, from day 3 to 11, and day 12 increasing or decreasing gene group transcription. As shown
to 26, respectively. Numbers at the Y-axis represent the averaged value of in Figure 3, clusters identified as upregulated in time-interval 1
samples expression inside each time interval. Continuous lines represent
have low average connectivity. Figure 4 shows the PPI network
significant alterations on the expression level when compared with the
previous time interval (pairwise t-test, corrected by FDR with p-value ≤ 0.01). of each cluster identified as upregulated in time-interval 1 and,
Dotted lines represent alterations considered non-significant. Lines started except by a few nodes in cluster 1 and cluster 4, the nodes of
and finished with a squared dot represent markers that present significant upregulated clusters are poorly connected. We then performed
changes detected by pairwise t-test between time-interval zero and time- Gene Ontology enrichment analysis of each upregulated cluster.
interval 2.
Transcriptogramer R/Bioconductor package implements a Gene

FIGURE 2 | Transcriptogram and connectivity diagram of time-intervals 1 and 2. (A, C) The transcriptogram of time-interval 1 (day 3 to 11, n = 9) and time-interval
2 (day 12 to 26, n = 15), respectively, from control and treated samples. The X-axis represents gene position, and the Y-axis the relative expression. The solid
black line represents control expression average, and the solid gray line represents the lead treatment expression average relative to control. Colored lines highlight
the groups of differentially expressed genes that compose the 11 identified clusters. Both transcriptograms were performed using radius = 80, and differentially
expressed clusters were selected with p-value ≤ 0.001. Clusters are represented as PPI networks and were functionally characterized by GO biological process
enrichment. Colored bars between transcriptograms illustrate the area occupied by each cluster on the X-axis. Top bar indicates the time-interval 1 clusters,
sequentially enumerated from left to right. Bottom bar indicates the time-interval 2 clusters identified by the correspondent former cluster number and color. Clusters
that arise only on time-interval 2 or embrace two or more clusters from time-interval 1 are labeled with capital letters and identified with a new color or the same
color of the former cluster with greater overlapping area, respectively. (B, D) The average network connectivity from time-interval 1 differentially expressed clusters.
The area occupied by each cluster in the circumference is proportional to the cluster average connectivity in the subnetwork (ratio between the number of cluster
connections and the number of cluster nodes). Clusters are identified by the same labels and colors used in (A) and (C). Colored lines on the circle perimeter
represent the area occupied by each cluster, normalized by the total number of cluster component interactions. Colored lines inside the circle illustrate the inner and
outer average connectivity, defined as the ratio between the number of all cluster protein connections and the number of proteins of each cluster. Lines connecting
the same cluster represent inner connectivity (connections among genes belonging to the same cluster), while lines connecting different clusters represent outer
connectivity. Node connections not included at any clusters are not represented.
Ontology enrichment analysis. For each differentially expressed transport. Cluster 4 had 19 enriched terms, which were related
cluster, we calculated the distance, all-against-all, of significantly with cellular adhesion and signaling transduction, especially
enriched GO terms. The distance is based on the Jaccard Index GTP-mediated cascades. There was no GO term significantly
between any GO term pair. The resulting adjacency matrix is enriched associated with cluster 2. It is important to note that
then used to plot a hierarchical network of significantly enriched only cluster 1 GO terms were sufficiently associated with each
GO terms associated with each cluster identified as differentially other to form a hierarchical network. Accordingly, lead-induced
expressed in transcriptogram analysis (see the Materials and increase in gene expression in time-interval 1 seems not to be
Methods section for details). GO enrichment analysis of cluster 1 related to massive biochemical system modulation.
resulted in 37 significantly enriched GO terms. The terms were
associated with molecule biosynthesis, especially RNA molecules,
transcription, and gene expression regulation (Figure 4 and Functional Enrichment of Downregulated
Supplementary Table S1). The four cluster 3 enriched terms Clusters in Time-Interval 1
were related to transmembrane transport, protein complex Among the clusters identified as downregulated on time-
oligomerization, ion transport, and cellular potassium ion interval 1, clusters 7 to 10 represent large and highly

FIGURE 3 | Evolution of network average connectivity and number of nodes. Cluster labels refer to time-interval 2. Clusters labeled with numbers were identified in
time-intervals 1 and 2; merged clusters, as well as new clusters, were labeled with letters. Bar colors refer to cluster colors at time-intervals 1 and 2, respectively.
Stacked bar charts represent clusters of time-interval 1 merged on time-interval 2. Numbers at the X-axis identify the time interval. Red, blue, and gray bars denote
the expression status (upregulated clusters, downregulated clusters, and absent cluster at that time interval, respectively). Clusters 5 and 11 have both upregulated
and downregulated expression patterns on time-interval 2. Top panels show the number of nodes variation through time intervals. Bottom panels represent the
average connectivity (< k>) variation through time intervals.
connected networks (Figure 5). Terms significantly enriched GO terms. Time-interval 2 total interactome enlargement is
in those clusters include processes related to RNA metabolism followed by an increase in the total number of enriched GO
and gene expression regulation. GO enrichment analysis terms (Supplementary Figure S7). In the first time interval, the
of cluster 7 resulted in 242 enriched GO terms. Among majority of cluster terms are related to RNA biosynthesis and
them, there were terms related to chromatin organization, metabolism, pointing to an acute disturbance of transcriptional
DNA-damage cellular response, negative regulation of cell pattern induced by lead exposure. In the second time interval,
cycle, negative regulation of transcription processes, and it is difficult to identify a unique biological process affected,
negative regulation of gene expression. GO terms associated pointing to a massive alteration in cell metabolism.
with cluster 8 (43 terms) were related to RNA biosynthesis,
particularly of non-coding RNAs. Cluster 9 terms (25 at
total) were associated with mRNA processing and protein DISCUSSION
biosynthesis and transport. Similarly, cluster 10 was enriched
Lead poisoning effects are broad and affect several cellular
with terms related to protein biosynthesis, also having terms
systems (Neal and Guilarte, 2010; Mitra et al., 2017). Among
of RNA catabolism regulation (96 terms, at total).
them, neurological effects of lead poisoning are critical,
Clusters 5, 6, and 11 are characterized by the shifting
especially during development (Lidsky and Schneider, 2003;
expression between time intervals (Figure 2). PPI networks
Martino and Pluchino, 2006; White et al., 2007; Sanders et al.,
and enriched terms are presented in Supplementary Material
2009). According to our results, lead treatment was able to
Online (Supplementary Figure S6). GO enrichment analysis
strongly decrease the expression of several cellular systems in the
of cluster 5 resulted in 29 enriched GO terms associated with
first 11 days of treatment, resulting in a massive transcriptional
vesicle formation, transport, and exocytosis. GO enrichment
impairment observed at the end of the treatment. Lead is classified
analysis of cluster 6 resulted in 188 terms mainly related to
as a potent neurotoxin, interfering with specific brain areas and
cell cycle regulation and cytoskeleton organization. Finally,
neuronal pathways, being particularly harmful to children. Early
cluster 11 enrichment results in 20 terms related to oxidative
and chronic exposure, even to low lead levels, were associated
metabolism and ATP metabolic process. In contrast with
with brain damage, abnormal neurodevelopment, impairment of
upregulated clusters, downregulated cluster GO terms are
IQ levels, neuropsychological dysfunction, as well as behavioral
numerous (Supplementary Table S1). Additionally, enriched
disorders (Meyer et al., 2008; Schneider et al., 2012; Schnur
terms in each downregulated cluster are functionally related
and John, 2014). The results observed here suggest that lead
to each other inside the cluster, as can be observed by the GO
neurotoxicity during development could be associated with a
term hierarchical networks (Figure 5 and Supplementary
huge transcriptional impairment in developing nerve cells.
Figures S8–S17).
Cell differentiation requires strict regulation of gene
expression in order to ensure correct cell fate. Some studies
Functional Enrichment in Time-Interval 2 report lead exposure impact on embryonic stem cells
The massive alteration in transcription profile observed on time- differentiation (Huang and Schneider, 2004; Abdullah et al.,
interval 2 reflects on cluster network properties, especially the 2014; Senut et al., 2014). In a study involving embryonic stem
number of nodes and connectivity, and also in their enriched cells cultivated with lead acetate 10 μM for 5 days, the authors

FIGURE 4 | PPI network visualization and term dendrograms of upregulated clusters. Transcriptogram, cluster names, and cluster colors correspond to time-
interval 1. The circle over the transcriptogram indicates the highlighted upregulated clusters. PPI network nodes are represented as ellipsis. Upregulated cluster
networks are small and poorly connected, as it is possible to observe in network representation. Dendrograms refer to clusters enriched GO hierarchy. Dendrogram
node sizes are proportional to the number of terms held by each GO term. Dendrogram colors represent the normalized terms occupation rate, where darker
colors indicate the ratio between the number of cluster genes overlapping to the term genes and the number of term genes, normalized by the maximum ratio
found in the cluster.
observed significant proliferation inhibition in neurospheres the marker expression dynamics were not the same among
collected from rat brain regions (Huang and Schneider, 2004). treated and control (as shown in Supplementary Figures
In contrast, Senut et al. (2014) observed exposure to lead S4 and S5), which suggests that the differentiation process
acetate (0.4 and 1.9 μM) did not affect the differentiation was impaired in some extent.
of human embryonic stem cells (hESCs) to NPCs. However, Jiang and collaborators investigated the expression of specific
these authors showed that lead could change neuronal features genes to identify possible lead-associated disease development
such as neurites and branching when NPCs differentiate biomarkers. They found transcriptional alterations in genes
to neurons, while also altering the methylation pattern of involved with cell–cell signaling, cell development, and cell
genes involved in neurogenetic signaling pathways (Senut cycle, as well as response to stress and DNA damage. The
et al., 2014). The study conducted by Jiang and collaborators authors have described early modulation of cell cycle genes,
has investigated the effect of lead treatment in ES-NP cells neurotransmitter transport, and organelle fission. Later
during differentiation to neuronal cells to evaluate lead- modulated genes were associated to positive regulation of
caused neurotoxicity (Jiang et al., 2017). However, there is biological, metabolic, and immune system processes (Jiang
no assay in the original paper to assess the differentiation et al., 2017). Here, we investigate the same data from a systemic
process efficiency. We measured the expression of classical perspective, since transcriptogramer provides a global view of
NPCs and neuronal marker genes during the experiment to cellular metabolism by indicating functionally associated gene
evaluate the differentiation of ES-NP cells in neuronal cells. sets with altered expression in a given biological condition. The
Overall, the NPC markers decrease their expression and the approach used here allowed to globally assess the transcriptional
neuronal markers increased their expression throughout the impact of lead treatment in impairing entire biochemical
experiment in both control and treated cells. This pattern is systems, and therefore, our results are complementary to the
consistent with a successful differentiation process. However, results of Jiang and collaborators.

FIGURE 5 | PPI network visualization and terms dendrograms of downregulated clusters. Transcriptogram, clusters names, and cluster colors correspond to the
time-interval 1. The circle over the transcriptogram indicates the highlighted downregulated clusters. Nodes are the cluster’s relevant proteins but are not explicitly
represented. The confluence of edges identifies them. Dendrograms refer to the clusters enriched GO hierarchy. Node sizes are proportional to the numbers of
terms held by each GO term. Dendrogram colors represent the normalized term occupation rate, where darker colors indicate the ratio between the number of
cluster genes overlapping to the term genes and the number of term genes, normalized by the maximum ratio found in the cluster. Zoom areas on the dendrogram
show relevant terms based on the occupation rate index.
According to our results, lead treatment was able to decrease terms in cluster 8 are related to nucleotide metabolic process
entire cellular systems in time-interval 1 as denoted by the large and nucleotide salvage, especially purine-containing ribose-
and densely connected downregulated networks, representing phosphate nucleotides, such as ATP and GTP. It might reflect,
biological functions related to cell cycle regulation, chromatin to some extent, into impairment in energetic metabolism.
modification, cytoskeleton organization, RNA biosynthesis This illustrates lead interference in the various layers of gene
and transcription regulation, protein biosynthesis and protein expression regulation, from chromatin modifications to cellular
transport, vesicle formation, and exocytosis. When taking exocytosis. On the other hand, upregulated networks in time-
together, the downregulation of those biochemical systems interval 1 were composed of few nodes poorly connected. It
suggests deregulation in the vital cellular process, which could indicate an initial activation of systems, which will be
might result in a differentiation impairment. Several GO massively modulated later, as observed in time-interval 2.

Reinforcing that, several GO terms significantly enriched impairment during ES-NP cell differentiation, affecting
in time-interval 1 upregulated clusters were associated with several biochemical systems crucial to neurodevelopment,
transcriptional modulation, such as ion transport, GTP- such as mTOR signaling. The comprising in transcriptional
mediated cascades, and gene expression regulation itself. On homeostasis is compatible with neuronal and progenitor
the second time interval, upregulated cluster networks were marker expression dynamics. Therefore, it is reasonable
bigger and more connected when compared to time-interval 1, to assume that the normal development of neuronal cells
suggesting a late modulation of cellular processes mediated by could be impaired in lead intoxication, especially by
upregulated clusters. altering transcription homeostasis. However, it is difficult
Several biochemical systems that were downregulated in to determine the real consequence of lead in central nervous
time-interval 1 (e.g., cycle regulation, transcription regulation, system development in vivo, and further studies are needed to
protein biosynthesis, and nucleotide-phosphate metabolism) totally elucidate lead poisoning neuronal impact.
are still downregulated in time-interval 2. Considering time-
interval transition, both transcriptograms show an extensive
time-dependent alteration of ES-NP cell transcription DATA AVAILABILITY
pattern. Additionally, the behavior of neuronal marker
expression in control and treated samples reinforces those Data supporting this paper can be accessed through https://
observations. Four of the six neuronal markers (i.e., GAD1, github.com/LabBiosystemUFRN/LeadNPC
GAD2, NEUROD6, and TH) were not significantly altered in Datasets analyzed for this study can be found in the NCBI-
time-interval 1, while all the evaluated neuronal markers were Bioproject Accession PRJNA330909 (GEO: GSE84712) at https://
significantly altered in time-interval 2 (always comparing www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE84712.
treated samples with control samples). Interestingly, some of
them are upregulated in treated samples (RBFOX3 and TH),
while others are downregulated in treated samples (GAD1, AUTHOR CONTRIBUTIONS
GAD2, and NEUROD6). All the NPC markers evaluated are
downregulated in treated samples in time-interval 1, agreeing CR and IS performed the analysis and wrote the manuscript.
with the huge downregulation of several biochemical systems. DM collected the data and performed the analysis. RO and DI
One of the downregulated systems in time-interval 1 is the performed the analysis and organized the supplementary material.
mTOR signaling proteins (GO:0031929, Supplementary RD and RA designed the study and wrote the manuscript. All
Table S1) found in cluster 6. mTOR signaling cascade is authors reviewed and approved the final manuscript. CR and IS
related to neuronal and glial development, learning, memory, have contributed equally to this work.
and synaptic plasticity, being activated by the brain-derived
neurotrophic factor (BDNF) (Lipton and Sahin, 2014; Switon et
al., 2017). In a previous study, Neal et al. (2010) demonstrated FUNDING
that lead exposure decreased BDNF levels in hippocampal
neurons. The authors also observed that exogenous treatment This work has been financed by the governmental Brazilian agency
with BDNF applied to lead-exposed cells recovered presynaptic National Council for Scientific and Technological Development
protein levels and vesicular neurotransmitter release, which (CNPq, Portuguese: Conselho Nacional de Desenvolvimento
suggests a presynaptic mechanism for lead poisoning (Neal Científico e Tecnológico), grant 444856/2014-5. The scholarships
et al., 2010; Stansfield et al., 2012). Synaptic plasticity is affected were financed by governmental Brazilian agency Coordination
by lead during neurodevelopment and mTOR signaling for the Improvement of Higher Education Personnel (CAPES–
cascade could be a biological target to lead poisoning (White Portuguese: Coordenação de Aperfeiçoamento de Pessoal de
et al., 2007). However, the interaction of mTOR and BDNF Nível Superior).
signaling pathways in lead poisoning should be properly
tested in future studies, which might help to clarify lead
neurotoxicity mechanism. ACKNOWLEDGMENTS
The diverse and systemic effects of lead poisoning reflect
lead ability to impair several cellular components, such as The authors would like to thank the NPAD/UFRN for
receptors, membranes, and transcription factors, disturbing computational resources.
cell function (de Souza et al., 2018). Lead interference in gene
expression is reported in several studies, specially suggesting
gene expression disturbance as the underlying mechanism for SUPPLEMENTARY MATERIAL
lead neurotoxicity (Nihei and Guilarte, 1999; Bouton et al.,
2001; Nihei and Guilarte, 2001; Kasten-Jolly et al., 2011; The Supplementary Material for this article can be found online at:
Kasten-Jolly et al., 2012; Schneider et al., 2012). The results https://www.frontiersin.org/articles/10.3389/fgene.2019.00791/
showed here demonstrated a lead-induced transcriptional full#supplementary-material

REFERENCES Kasten-Jolly, J., Heo, Y., and Lawrence, D. A. (2011). Central nervous system
cytokine gene expression: modulation by lead. J. Biochem. Mol. Toxicol. 25,
Abdullah, M., Rahman, F. A., Gnanasegaran, N., Govindasamy, V., Abu 41–54. doi: 10.1002/jbt.20358
Kasim, N. H., and Musa, S. (2014). Diverse effects of lead nitrate on Kasten-Jolly, J., Pabello, N., Bolivar, V. J., and Lawrence, D. A. (2012).
the proliferation, differentiation, and gene expression of stem cells Developmental lead effects on behavior and brain gene expression in male and
isolated from a dental origin. Sci. World J. 2014, 1–12. doi: 10.1155/2014/ female BALB/cAnNTac mice. Neurotoxicology 33, 1005–1020. doi: 10.1016/j.
235941 neuro.2012.04.017
Baranowska-Bosiacka, I., Gutowska, I., Rybicka, M., Nowacki, P., and Chlubek, D. Khan, S., Stott, S. R., Chabrat, A., Truckenbrodt, A. M., Spencer-Dene, B.,
(2012). Neurotoxicity of lead. Hypothetical molecular mechanisms of Nave, K. A., et al. (2017). Survival of a novel subset of midbrain dopaminergic
synaptic function disorders. Neurol. Neurochir. Pol. 46, 569–578. doi: 10.5114/ neurons projecting to the lateral septum is dependent on neuroD proteins.
ninp.2012.31607 J. Neurosci. 37, 2305–2316. doi: 10.1523/JNEUROSCI.2414-16.2016
Bouton, C. M. L. S., Frelin, L. P., Forde, C. E., Godwin, H. A., and Pevsner, J. (2001). Lidsky, T. I., and Schneider, J. S. (2003). Lead neurotoxicity in children: basic
Synaptotagmin I is a molecular target for lead. J. Neurochem. 76, 1724–1735. mechanisms and clinical correlates. Brain 126, 5–19. doi: 10.1093/brain/
doi: 10.1046/j.1471-4159.2001.00168.x awg014
Castro, M. A., de Santiago, I., Campbell, T. M., Vaughn, C., Hickey, T. E., Ross, E., Lipton, J. O., and Sahin, M. (2014). The neurology of mTOR. Neuron 84, 275–291.
et al. (2016). Regulators of genetic risk of breast cancer identified by integrative doi: 10.1016/j.neuron.2014.09.034
network analysis. Nat. Genet. 48, 12–21. doi: 10.1038/ng.3458 Lutolf, S., Radtke, F., Aguet, M., Suter, U., and Taylor, V. (2002). Notch1 is required
Castro, M. A., Wang, X., Fletcher, M. N., Meyer, K. B., and Markowetz, F. (2012). for neuronal and glial differentiation in the cerebellum. Development 129,
Reder: R/bioconductor package for representing modular structures, nested 373–385.
networks and multiple levels of hierarchical associations. Genome Biol. 13, R29. Lê, S., Josse, J., and Husson, F. (2008). Factominer: An r package for multivariate
doi: 10.1186/gb-2012-13-4-r29 analysis. J. Stat. Softw. Artic. 25, 1–18. doi: 10.18637/jss.v025.i01
Chambers, S. M., Fasano, C. A., Papapetrou, E. P., Tomishima, M., Sadelain, M., Martino, G., and Pluchino, S. (2006). The therapeutic potential of neural stem
and Studer, L. (2009). Highly efficient neural conversion of human ES and iPS cells. Nat. Rev. Neurosci. 7, 395–406. doi: 10.1038/nrn1908
cells by dual inhibition of SMAD signaling. Nat. Biotechnol. 27, 275–280. doi: Mathews, K. J., Allen, K. M., Boerrigter, D., Ball, H., Shannon Weickert, C., and
10.1038/nbt.1529 Double, K. L. (2017). Evidence for reduced neurogenesis in the aging human
Cossette, M., Levesque, D., and Parent, A. (2005). Neurochemical characterization hippocampus despite stable stem cell markers. Aging Cell 16, 1195–1199. doi:
of dopaminergic neurons in human striatum. Parkinsonism Relat. Disord. 11, 10.1111/acel.12641
277–286. doi: 10.1016/j.parkreldis.2005.02.008 Meyer, P. A., Brown, M. J., and Falk, H. (2008). Global approach to reducing
de Souza, I. D., de Andrade, A. S., and Dalmolin, R. J. S. (2018). Lead-interacting lead exposure and poisoning. Mutat. Res. Rev. Mutat. Res. 659, 166–175. doi:
proteins and their implication in lead poisoning. Crit. Rev. Toxicol. 0, 1–12. doi: 10.1016/j.mrrev.2008.03.003
10.1080/10408444.2018.1429387 Mitra, P., Sharma, S., Purohit, P., and Sharma, P. (2017). Clinical and molecular
Finkelstein, Y., Markowitz, M. E., and Rosen, J. F. (1998). Low-level lead-induced aspects of lead toxicity: an update. Crit. Rev. Clin. Lab. Sci. 54, 506–528. doi:
neurotoxicity in children: an update on central nervous system effects. Brain 10.1080/10408363.2017.1408562
Res. Rev. 27, 168–176. doi: 10.1016/S0165-0173(98)00011-3 Morais, D. A. A., Almeida, R. M. C., and Dalmolin, R. J. S. (2019).
Fletcher, M. N., Castro, M. A., Wang, X., de Santiago, I., O’Reilly, M., Chin, S. F., Transcriptogramer: an R/Bioconductor package for transcriptional analysis
et al. (2013). Master regulators of FGFR2 signalling and breast cancer risk. Nat. based on protein–protein interaction. Bioinformatics 1–2. doi: 10.1093/
Commun. 4, 2464. doi: 10.1038/ncomms3464 bioinformatics/btz007
Flora, G., Gupta, D., and Tiwari, A. (2012). Toxicity of lead: a review with recent Neal, A. P., and Guilarte, T. R. (2010). Molecular neurobiology of lead (Pb(2+)):
updates. Interdiscip. Toxicol. 5, 47–58. doi: 10.2478/v10102-012-0009-2 effects on synaptic function. Mol. Neurobiol. 42, 151–160. doi: 10.1007/
Ghering, A. B., Jenkins, L. M. M., Schenck, B. L., Deo, S., Mayer, R. A., Pikaart, s12035-010-8146-0
M. J., et al. (2005). Spectroscopic and functional determination of the Neal, A. P., Stansfield, K. H., Worley, P. F., Thompson, R. E., and Guilarte, T. R.
interaction of Pb 2+ with GATA proteins. J. Am. Chem. Soc. 127, 3751–3759. (2010). Lead exposure during synaptogenesis alters vesicular proteins and
doi: 10.1021/ja0464544 impairs vesicular release: potential role of NMDA receptor-dependent BDNF
Gu, Z., Gu, L., Eils, R., Schlesner, M., and Brors, B. (2014). Circlize implements signaling. Toxicol. Sci. 116, 249–263. doi: 10.1093/toxsci/kfq111
and enhances circular visualization in r. Bioinformatics 30, 2811–2812. doi: Nihei, M. K., Desmond, N. L., McGlothan, J. L., Kuhlmann, A. C., and
10.1093/bioinformatics/btu393 Guilarte, T. R. (2000). N-methyl-D-aspartate receptor subunit changes are
Guilarte, T. R., and McGlothan, J. L. (1998). Hippocampal NMDA receptor mRNA associated with lead-induced deficits of long-term potentiation and spatial
undergoes subunit specific changes during developmental lead exposure. Brain learning. Neuroscience 99, 233–242. doi: 10.1016/S0306-4522(00)00192-5
Res. 790, 98–107. doi: 10.1016/S0006-8993(98)00054-7 Nihei, M. K., and Guilarte, T. R. (1999). NMDAR-2A subunit protein
Gusel’nikova, V. V., and Korzhevskiy, D. E. (2015). NeuN as a neuronal nuclear expression is reduced in the hippocampus of rats exposed to Pb2+
antigen and neuron differentiation marker. Acta Naturae 7, 42–47. doi: during development. Brain Res. Mol. Brain Res. 66, 42–49. doi: 10.1016/
10.32607/20758251-2015-7-2-42-47 S0169-328X(99)00005-4
Huang, F., and Schneider, J. (2004). Effects of lead exposure on proliferation Nihei, M. K., and Guilarte, T. R. (2001). Molecular changes in glutamatergic
and differentiation of neural stem cells derived from different regions synapses induced by Pb2+: association with deficits of LTP and spatial learning.
of embryonic rat brain. Neurotoxicology 25, 1001–1012. doi: 10.1016/j. Neurotoxicology 22, 635–643. doi: 10.1016/S0161-813X(01)00035-3
neuro.2004.03.010 Ordemann, J. M., and Austin, R. N. (2016). Lead neurotoxicity: exploring the
Husson, F., Josse, J., and Pagès, J. (2010). “Principal component methods- potential impact of lead substitution in zinc-finger proteins on mental health.
hierarchical clustering-partitional clustering: why would we need to choose for Metallomics 8, 579–588. doi: 10.1039/C5MT00300H
visualizing data,” in Technical Report-Agrocampus. A. C. ouest (Rennes: Applied Pertea, M., Kim, D., Pertea, G. M., Leek, J. T., and Salzberg, S. L. (2016). Transcript-
mathematics department), 17. level expression analysis of RNA-Seq experiments with hisat, stringtie and
Jiang, P., Hou, Z., Bolin, J. M., Thomson, J. A., and Stewart, R. (2017). RNA-seq ballgown. Nat. Protoc. 11, 1650. doi: 10.1038/nprot.2016.095
of human neural progenitor cells exposed to lead (Pb) reveals transcriptome Reddy, G. R., and Zawia, N. H. (2000). Lead exposure alters Egr-1 DNA-binding
dynamics, splicing alterations and disease risk associations. Toxicol. Sci. in the neonatal rat brain. Int. J. Dev. Neurosci. 18, 791–795. doi: 10.1016/
159, 251–265. doi: 10.1093/toxsci/kfx129 S0736-5748(00)00048-4
Kaneko, Y., Sakakibara, S., Imai, T., Suzuki, A., Nakamura, Y., Sawamoto, K., et al. Richardt, G., Federolf, G., and Habermann, E. (1986). Affinity of heavy metal ions
(2000). Musashi1: an evolutionally conserved marker for CNS progenitor cells to intracellular Ca2+-binding proteins. Biochem. Pharmacol. 35, 1331–1335.
including neural stem cells. Dev. Neurosci. 22, 139–153. doi: 10.1159/000017435 doi: 10.1016/0006-2952(86)90278-9

Ritchie, M. E., Phipson, B., Wu, D., Hu, Y., Law, C. W., Shi, W., et al. (2015). Limma Suter, D. M., Tirefort, D., Julien, S., and Krause, K. H. (2009). A Sox1 to Pax6
powers differential expression analyses for RNA-sequencing and microarray switch drives neuroectoderm to radial glia progression during differentiation
studies. Nucleic Acids Res. 43, e47. doi: 10.1093/nar/gkv007 of mouse embryonic stem cells black. Stem Cells 27, 49–58. doi: 10.1634/
Ronis, M. J. J., Aronson, J., Gao, G. G., Hogue, W., Skinner, R. A., Badger, T. M., stemcells.2008-0319
et al. (2001). Skeletal effects of developmental lead exposure in rats. Toxicol. Sci. Switon, K., Kotulska, K., Janusz-Kaminska, A., Zmorzynska, J., and Jaworski, J.
62, 321–329. doi: 10.1093/toxsci/62.2.321 (2017). Molecular neurobiology of mTOR. Neuroscience 341, 112–153. doi:
Rybarczyk-Filho, J. L., Castro, M. A. A., Dalmolin, R. J. S., Moreira, J. C. F., 10.1016/j.neuroscience.2016.11.017
Brunnet, L. G., and de Almeida, R. M. C. (2011). Towards a genome-wide White, L. D., Cory-Slechta, D. A., Gilbert, M. E., Tiffany-Castiglioni, E.,
transcriptogram: the Saccharomyces cerevisiae case. Nucleic Acids Res. 39, Zawia, N. H., Virgolini, M., et al. (2007). New and evolving concepts in the
3005–3016. doi: 10.1093/nar/gkq1269 neurotoxicology of lead. Toxicol. Appl. Pharmacol. 225, 1–27. doi: 10.1016/j.
Sanders, T., Liu, Y., Buchner, V., and Tchounwou, P. B. (2009). Neurotoxic effects taap.2007.08.001
and biomarkers of lead exposure: a review. Rev. Environ. Health 24, 15–45. doi: White, R. B., and Thomas, M. G. (2012). Moving beyond tyrosine hydroxylase
10.1515/REVEH.2009.24.1.15 to define dopaminergic neurons for use in cell replacement therapies for
Schneider, J. S., Mettil, W., and Anderson, D. W. (2012). Differential effect of Parkinson’s disease. CNS Neurol. Disord. Drug Targets 11, 340–349. doi:
postnatal lead exposure on gene expression in the hippocampus and frontal 10.2174/187152712800792758
cortex. J. Mol. Neurosci. 47, 76–88. doi: 10.1007/s12031-011-9686-0 Wiese, C., Rolletschek, A., Kania, G., Blyszczuk, P., Tarasov, K. V., Tarasova, Y.,
Schnur, J., and John, R. M. (2014). Childhood lead poisoning and the new centers et al. (2004). Nestin expression—a property of multi-lineage progenitor cells?
for disease control and prevention guidelines for lead exposure. J. Am. Assoc. Cell. Mol. Life Sci. 61, 2510–2522. doi: 10.1007/s00018-004-4144-6
Nurse Pract. 26, 238–247. doi: 10.1002/2327-6924.12112 Zawia, N., Sharan, R., Brydie, M., Oyama, T., and Crumpton, T. (1998). Sp1 as
Selvaraj, V., Jiang, P., Chechneva, O., Lo, U. G., and Deng, W. (2012). Differentiating a target site for metal-induced perturbations of transcriptional regulation
human stem cells into neurons and glial cells for neural repair. Front. Biosci. of developmental brain gene expression. Dev. Brain Res. 107, 291–298. doi:
(Landmark Ed.) 17, 65–89. doi: 10.2741/3916 10.1016/S0165-3806(98)00023-6
Senut, M.-C., Sen, A., Cingolani, P., Shaik, A., Land, S. J., and Ruden, D. M. (2014). Lead
exposure disrupts global DNA methylation in human embryonic stem cells and alters Conflict of Interest Statement: The authors declare that the research was
their neuronal differentiation. Toxicol. Sci. 139, 142–161. doi: 10.1093/toxsci/kfu028 conducted in the absence of any commercial or financial relationships that could
Shannon, P. T., Grimes, M., Kutlu, B., Bot, J. J., and Galas, D. J. (2013). Rcytoscape: be construed as a potential conflict of interest.
tools for exploratory network analysis. BMC Bioinformatics 14, 217. doi:
10.1186/1471-2105-14-217 Copyright © 2019 Reis, de Souza, Morais, Oliveira, Imparato, de Almeida
Stansfield, K. H., Richard Pilsner, J., Lu, Q., Wright, R. O., and Guilarte, T. R. (2012). and Dalmolin. This is an open-access article distributed under the terms of
Dysregulation of BDNF-TrkB signaling in developing hippocampal neurons the Creative Commons Attribution License (CC BY). The use, distribution
by Pb 2+: implications for an environmental basis of neurodevelopmental or reproduction in other forums is permitted, provided the original author(s)
disorders. Toxicol. Sci. 127, 277–295. doi: 10.1093/toxsci/kfs090 and the copyright owner(s) are credited and that the original publication in
Sun, X., Tian, X., Tomsig, J. L., and Suszkiw, J. B. (1999). Analysis of differential this journal is cited, in accordance with accepted academic practice. No
effects of Pb2+ on protein kinase C isozymes. Toxicol. Appl. Pharmacol. 156, use, distribution or reproduction is permitted which does not comply with
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44
Discussão
O chumbo é classificado como uma toxina potente, cujo envenenamento causa efeitos
amplos e diversos. Além de afetar uma variada gama de tecidos, o chumbo age em diferentes
nı́veis do metabolismo celular, afetando muitos sistemas moleculares. Seus efeitos mais
significativos ocorrem no CNS, onde interfere em áreas cerebrais bastante especı́ficas e em
vias neuronais importantes. Os riscos do envenenamento por chumbo causam preocupação
especial quando ocorrem em crianças e mulheres grávidas, já que nestas populações a taxa de
absorção deste metal é maior e seus efeitos são particularmente perturbadores sobre um sistema
nervoso em desenvolvimento [53, 54, 5]. Apesar de ser mundialmente aceito como padrão para
inı́cio de ações de combate à intoxicação, nem mesmo o valor de 5µg/dL de sangue adotado
pelo Center of Disease Control americano (CDC) [55] pode ser considerado seguro, uma vez
que a exposição precoce e crônica ao chumbo, mesmo que a nı́veis bastante reduzidos, já foi
associada a danos cerebrais, neurodesenvolvimento anormal, comprometimento dos nı́veis de QI,
disfunção neuropsicológica e transtornos comportamentais [56, 57, 58]. Além disso, os efeitos
do P b2+ sobre a cognição e padrões comportamentais em crianças podem ser considerados
permanentes. Apesar das terapias quelantes à base de EDTA conseguirem reduzir a carga
corporal do metal, elas não corrigem os distúrbios já causados ao cérebro durante perı́odos
considerados crı́ticos do seu desenvolvimento, cujas consequências podem estar correlacionadas
a comportamentos anti-sociais, violência e redução no volume cerebral em adultos [59]. Os
efeitos sistêmicos do envenenamento por chumbo refletem sua competência na interação com
muitos componentes capazes de causar distúrbios na função celular [23].
Estudos anteriores envolvendo diferenciação de células-tronco embrionárias demonstraram a
influência deste metal sobre o metabolismo, alterando as caracterı́sticas celulares em diferentes
nı́veis. Foram reportadas alterações fenotı́picas na morfologia, onde os neurônios resultantes
de NPCs tratadas com concentrações maiores que 1, 9µM de acetato de chumbo por um
perı́odo de 11 a 19 dias exibiram um menor desenvolvimento neurı́tico, geralmente adotando
uma forma unipolar com pequena ramificação dendrı́tica e um longo axônio. Alterações
epigenéticas significativas também foram registradas, sugerindo a vulnerabilidade de células
neuronais em diferenciação à ação do chumbo. Genes essenciais à neurogênese tiveram seus
padrões de metilação alterados, como no caso do PLXNA4 e do NEUROG1, que apresentaram
hipermetilação e hipometilação, respectivamente. Em tais estudos foram, ainda, observadas
expressivas reduções nas taxas de proliferação de células tronco neurais quando tratadas com
P b2+ [60, 61].
No presente trabalho, observou-se que os efeitos sistêmicos do chumbo afetaram tanto o
processo de diferenciação celular quanto outros processos celulares diversos, diferindo de forma
significativa com o decorrer do tempo de tratamento. Houve como efeito precoce (perı́odo
entre os dias 3 e 11 do tratamento, Time-Interval 1 ou T 1) uma grande redução nos valores
Discussão 45
de expressão de vários sistemas. Num segundo momento (perı́odo entre os dias 12 e 26 do

tratamento, Time-Interval 2 ou T 2) houve um comprometimento maciço do perfil transcricional
das células intoxicadas, tornando evidente os efeitos cumulativos de uma exposição prolongada
ao metal, sugerindo um comprometimento global do desenvolvimento das células nervosas
decorrente da neurotoxicidade do chumbo.
Efeitos do chumbo sobre a diferenciação celular

A diferenciação celular é um mecanismo complexo e delicado que dependente de uma
expressão gênica bastante especı́fica, a qual poderia ser alterada pela interferência do chumbo.
A fim de constatar a efetiva diferenciação neuronal, elegemos quatro marcadores para NPC e seis
marcadores para células neuronais. Nossos resultados mostram nı́veis de expressão compatı́veis
com um processo de diferenciação N P C 7→ N eurônio, onde os nı́veis de expressão dos
marcadores de NPC decaem e os nı́veis de expressão de marcadores Neuronais crescem com
o decorrer do tempo, tanto no tratamento quanto no controle. Entretanto, a dinâmica da
expressão de tais marcadores difere significativamente.
Comparando-se os marcadores NPC do controle e tratado no tempo 0 (perı́odo entre os
dias 0 e 2 do tratamento ou T 0) nota-se uma expressão média bastante similar em todos
os quatro marcadores. De forma geral, houve no intervalo T 0 7→ T 2 um declı́nio suave e
constante nos nı́veis de expressão dos marcadores NPC do controle. Em contraste, o tratamento
apresentou uma queda abrupta nos nı́veis de expressão em três dos quatro marcadores no
intervalo T 0 7→ T 1, seguida de um platô no intervalo T 1 7→ T 2. Nossos dados mostram que
esta queda abrupta nos nı́veis de expressão de todos os marcadores ocorre logo nos primeiros
dias da exposição ao chumbo. Este comportamento encontra respaldo na literatura, onde é
sugerido que o gene SOX1 tem sua atividade silenciada pela inibição induzida por P b2+ de
vias metabólicas mediadas por GABA [61]. Também encontram-se relatos de uma redução
significativa na expressão do MSI1 em células expostas ao chumbo [60].
Já os marcadores neuronais apresentaram um comportamento menos uniforme, apesar de
compatı́veis com o processo de diferenciação em curso. De maneira geral, todos seis marcadores
apresentaram aumento considerável em seus nı́veis de expressão no intervalo T 0 7→ T 1, seguido
de um aumento menos significativo no intervalo T 1 7→ T 2, tanto no controle quanto no
tratamento. Observados dia a dia, podemos notar um aumento abrupto na expressão de
todos os marcadores logo nos primeiros dias de tratamento, em contraste com a evolução
dos marcadores do controle, que apresentaram um aumento mais gradual e uniforme. Este
fato, aliado à queda prematura na expressão dos marcadores NPC, nos sugere uma aceleração
na diferenciação neuronal provocada pelo chumbo, o que nos permitiria especular sobre duas
hipóteses:
1. A proteı́na transmembrana NOTCH1 tem um papel já bem descrito como reguladora da
diferenciação celular N P C 7→ N eurônio. Ativada pela proteı́na transmembrana Delta1
Discussão 46
Figura 12 – Mecanismo de bloqueio na diferenciação de NPC vizinhas causado pelo inı́cio

do processo de diferenciação de um neurônio.
de células vizinhas em diferenciação, dispara a produção de repressores de transcrição,

como HES1 e HES5, que inibirão a expressão de genes proneurais e do próprio Delta1.
Desta forma, um neurônio em diferenciação impede que células progenitoras neurais
vizinhas também entrem em diferenciação (Figura 12). Como consequência, uma queda
na atividade de sinalização Notch1 pode estar associada à diferenciação generalizada e
prematura de um grande número de neurônios simultaneamente [62], mediada por uma
hipotética interferência do P b2+ na ligação Delta1 7→Notch1 ou na via de sinalização
deste último.
2. O zinc-finger SP1 é um fator de transcrição capaz de conectar-se a motivos de ligação de

diversos promotores e, como tal, está sujeito a alterações em sua conformação, induzido
pelo chumbo. Esta alteração pode potencializar sua ligação ao DNA, superativando
mecanismos relacionados ao fator de crescimento NGF, acelerando, assim, a diferenciação
celular durante as fases iniciais deste processo [63].
Efeitos precoces – Processos regulados negativamente

Observou-se que no Time-Interval 1 diversos processos metabólicos vitais foram regulados
negativamente como consequência da exposição das NPCs ao P b2+ , levando a uma redução
na atividade de uma série de sistemas complexos, compostos por redes de interação grandes e
densamente conectadas, envolvendo cerca de 82% dos nós contidos em clusteres neste intervalo
de tempo e cerca de 29% das 15.154 proteı́nas utilizadas pelo transcriptogramer. A depleção
Discussão 47
no Time-Interval 1 de atividades como regulação do ciclo celular e modificação de cromatina

(cluster 7), regulação da biossı́ntese de RNA e transcrição de RNAs não codantes (cluster 8),
transcrição de RNA, processamento de mRNA, biossı́ntese e transporte de proteı́nas (cluster 9),
biossı́ntese de peptı́deos e catabolismo de mRNA (cluster 10) indica que os efeitos sistêmicos do
chumbo afetam de forma indiscriminada a homeostase transcricional. Até mesmo mecanismos
do metabolismo energético, como processos de resgate e processamento de nucleotı́deos ATP
e GTP (cluster 11), foram afetados significativamente. Tais alterações sugerem um possı́vel
comprometimento de toda a atividade neuronal das células nervosas recém diferenciadas, sendo
que boa parte dos fenômenos observados podem ser associados à habilidade que o chumbo
tem em mimetizar ı́ons Ca2+ e Zn2+ .
O zinco atua como um potente inibidor da abertura de canais de cálcio dependentes
de voltagem [30]. Assim, a substituição P b2+ Zn2+ resulta em diminuição nos nı́veis
intracelulares de cálcio com consequências diretas nas cascatas de sinalização de mensageiros
secundários Ca2+ dependentes, na transdução de sinal intracelular, na expressão gênica e
na transcrição. Quando a substituição P b2+ Zn2+ ocorre em fatores de transcrição zinc-
fingers, tal proteı́na tem o reconhecimento do seu domı́nio de ligação ao DNA modificado
devido a uma mudança conformacional, impedindo ou potencializando sua ação, e como
consequência, alterando a forma de desempenho de suas funções, muitas delas relativas à
replicação, transcrição e tradução do DNA. Há fortes evidências de que o chumbo compete
pelo local de ligação do zinco nas proteı́nas SP1, EGR-1, GATA-1 e TFIIIA, alterando tempo
e afinidade de ligação ao DNA [64, 65, 23]. Além disso, a natural afinidade do P b2+ por
fatores de transcrição contendo zinc-fingers, abre espaço para muitas outras interações ainda
desconhecidas, mas que podem impactar de inúmeras formas na regulação da expressão gênica.
Não menos importante é a capacidade do chumbo de substituir ı́ons Ca2+ nas metalo-
proteı́nas, como a calmodulina e a PKC. Estas proteı́nas já estão bem descritas como elementos
chave em vias metabólicas envolvidas na plasticidade sináptica e na consolidação da memória em
neurônios [66, 67]. O chumbo também pode interferir em muitas outras cascatas de sinalização
e mecanismos de transdução dependentes de cálcio, como vias metabólicas envolvendo cAMP
e proteı́na G [68, 69], dando àquele metal a predisposição de perturbar inúmeros sistemas
celulares, como relatado em diversos estudos que associam o distúrbio na expressão gênica a
um mecanismo celular subjacente à neurotoxicidade do chumbo [70, 71, 72, 73, 74, 58].
Os efeitos da intoxicação por chumbo sobre o equilı́brio redox já são bem conhecidos e
têm como consequências danos à membrana celular e ao DNA por conta do estresse oxidativo
[23]. Paradoxalmente, nota-se que o cluster 7 também registra um enriquecimento de termos
relacionados à reposta celular ao dano no DNA, cujas funções encontram-se depletadas. Este
fato, porém, já encontra antecedentes na literatura, cuja causa pode estar correlacionada à
competição do P b2+ com ı́ons de zinco essenciais às polimerases de DNA, além de outras
proteı́nas ligadas ao reparo por excisão que contêm motivos de zinc-fingers em seus domı́nios
de ligação [75, 76].
Discussão 48
Outro estudo de expressão gênica realizado em camundongos destacou a regulação positiva

de citocinas e fatores de transcrição relacionados à resposta imune, sugerindo que o chumbo
poderia desencadear processos inflamatórios no tecido cerebral [72]. Curiosamente, todos os
termos relacionados à resposta imune enriquecidos no Time-Interval 1 pertenciam a clusteres
regulados negativamente (clusteres 6, 7 e 5, nesta ordem de importância).
Alguns clusteres regulados negativamente no primeiro intervalo de tempo apresentaram um
comportamento diferente dos demais no transcurso do experimento, sofrendo uma inversão de
expressão no Time-Interval 2, indo de downregulated a upregulated, como no caso do cluster
6, ou assumindo uma conformação onde uma parte do cluster assumiu valores upregulated e
uma segunda parte continuou exibindo regulação negativa (clusteres 5 e 11). Dentre estes,
nos chamam a atenção os clusteres 5 e 6, já que apresentaram enriquecimento de GOs em
termos bastante correlacionados à função neuronal propriamente dita. No cluster 5, apesar da
quantidade de nós apresentada ser relativamente pequena (4,98% dos nós em clusteres no T 1),
estes mostraram-se bem conectados entre si, também exibindo uma quantidade considerável
de nós conectados aos clusteres 6 e 7. Embora apresentasse somente 29 termos enriquecidos,
estes encontravam-se bem organizados e correlacionados a processos de formação e transporte
de vesı́culas e exocitose em sua maioria. Já o cluster 6 apresentou uma grande quantidade
de nós (16,85% dos nós em clusteres no T 1), densamente conectados e enriquecido em
188 termos, dentre os quais podemos destacar os termos relacionados à polimerização de
microtúbulos, organização de citoesqueleto e regulação e sinalização do mTOR. Tomados
em conjunto, os termos dos clusteres 5 e 6 podem ser facilmente associados a processos
biológicos dependentes de cálcio e zinco caracterı́sticos de um neurônio em desenvolvimento,
como crescimento e especificação axonal, interação entre astrócitos e neurônios, e liberação
de neurotransmissores. Indiretamente podem ser relacionados a processos de excitabilidade
neuronal, plasticidade sináptica, potenciação de longa duração, memória e aprendizagem,
particularmente no caso do mTOR, que participa da montagem dos complexos proteicos
mTORC1 e mTORC2. Uma das funções principais do mTORC2 é regular a polimerização de
actina. Fosforilando quinases como Akt, SGK1 e PKC, participa de processos que regulam a
sobrevivência, crescimento e migração celular. No CNS desempenha papel importante, estando
relacionado a processos de neurogênese, crescimento, regeneração e mielinização axonal,
bem como ao desenvolvimento de dendritos. Quando desregulado, compromete processos
interneuronais, como plasticidade sináptica, memória e cognição, podendo estar associado a
doenças psiquiátricas como depressão, retardo mental, esquizofrenia, doença de Parkinson
e doença de Alzheimer, entre outras [77]. Estudos em camundongos demonstraram que a
redução do mTORC2 provoca o desenvolvimento de neurônios disfuncionais, menores e de
morfologia anormal, além de uma redução do volume encefálico. O funcionamento neuronal
anormal foi associado à inativação das vias metabólicas do Akt em neurônios dopaminérgicos,
e as alterações morfológicas relacionadas com a não ativação da PKC e de proteı́nas a sua
jusante, como GAP-43 e MARCK2, fundamentais para a formação do citoesqueleto de actina
Discussão 49
[78]. Outros estudos ainda correlacionam o fator BDNF, responsável pela ativação das cascatas
de sinalização do gene mTOR, ao desenvolvimento neuronal, aprendizado e memória [79, 80],
onde foi demonstrado que o chumbo reduz as quantidades de BDNF disponı́vel em neurônios,
com impactos nos nı́veis de proteı́nas pré-sinápticas e liberação de neurotransmissores [81, 82]
Efeitos precoces – Processos regulados positivamente

Também foi constatado que no Time-Interval 1 os clusteres de 1 a 4 foram regulados
positivamente. Em geral foram clusteres que apresentaram poucos nós, representando cerca
de 18% dos nós contidos em todos os clusteres e 6,4% do total de proteı́nas em nossa PPI.
Este aumento na expressão em relação ao controle ocorreu em processos biológicos ligados
à modulação de transcrição, expressão gênica e biossı́ntese molecular de RNA (cluster 1),
transporte de ı́ons, transporte transmembrana e oligomerização de proteı́nas (cluster 3), adesão
celular e à transdução de sinais mediada por GTP (cluster 4). Todos eles apresentaram um
enriquecimento de ontologia muito pequeno, ou mesmo nulo, como no caso do cluster 2, com
termos pouco correlacionados. Isso pode ser um indicador de que, neste primeiro momento,
ocorreu apenas a ativação de pequenos grupos de genes ou mesmo genes individuais, não
havendo modulação de sistemas inteiros. Observando-se o perfil transcricional da mesma área
no Time-Interval 2, onde os clusteres correlatos sofreram ampliações consideráveis, pode-se
inferir que tais ativações mediaram o inı́cio de um processo de modulação dos sistemas regulados
positivamente em um segundo momento, como resposta adaptativa tardia à intoxicação pelo
chumbo.
Efeitos do chumbo sobre o desenvolvimento do CNS

Embora o presente estudo não traga evidencias diretas de prejuı́zo a funções superiores do
CNS, poderı́amos especular que a depleção de todos estes grandes sistemas, quando tomados
em conjunto, poderia indicar um comprometimento na interação entre neurônios já nos primeiros
dias de intoxicação. A ação do chumbo sobre metaloproteı́nas e fatores de transcrição no tecido
nervoso central tem sido associada a dificuldades de aprendizagem e consolidação de memória
[83, 84], havendo indı́cios de correlação entre a intoxicação deste tecido e o aparecimento de
doenças neurodegenerativas como as doenças de Parkinson e Alzheimer [65, 23].
Interferindo na regulação das vias glutamatérgicas, o P b2+ causa a redução na liberação de
glutamato (Glu), considerado o principal neurotransmissor excitatório do CNS [85]. Responsável
pela transmissão do impulso nervoso em até metade das sinapses cerebrais [86], o glutamato
também atua como excitador dos receptores ionotrópicos NMDAR. Estes possuem papel
fundamental no bom funcionamento do neurônio, permitindo o trânsito de N a+ e Ca2+ para
dentro da célula e de K + para fora [87], estando intimamente relacionados à plasticidade
sináptica dependente de atividade. A interação do chumbo com receptores NMDAR é bem
Discussão 50
conhecida, havendo relatos de perturbação no neurodesenvolvimento associada ao chumbo em

modelos animais por diferentes modos. Isto se dá porque, além da sua influência pré-sináptica
já descrita, o P b2+ pode substituir o Zn2+ na pós-sinapse, onde este último atua como um
antagonista bastante especı́fico nas subunidades GluN1/GluN2A dos receptores glutamatérgicos
NMDAR. Estudos relatam que o ı́on de chumbo não apenas inibe a abertura do canal do
receptor NMDAR na fenda sináptica, mas também é capaz de alterar a dinâmica da composição
da subunidade do receptor durante o desenvolvimento do cérebro [88, 73, 89]. Isso pode
ser particularmente relevante já que durante as fases iniciais do desenvolvimento do CNS, os
NMDARs trocam sua composição de subunidades, partindo de uma predominância de GluN2B
para um estado final onde as subunidades GluN2A, possuidoras de uma maior afinidade com o
zinco, são maioria. Ocorrendo em todo o CNS, esta troca coincide com o perı́odo de maturação
da sinapses e da aquisição de habilidades de aprendizado [87]. Este fato poderia explicar alguns
dos efeitos permanentes da substituição P b2+ Zn2+ durante a infância.
Sabe-se também que os zinc-fingers SP1 e o EGR-1 estão intimamente associados ao
desenvolvimento cerebral, também podendo ser correlacionados à doença de Alzheimer, doença
de Parkinson e esquizofrenia. O EGR-1 é um fator de transcrição cuja interação com o chumbo
já é bem estudada em vias neuronais. Sua expressão é estimulada por fatores de crescimento,
atividade neuronal, hormônios e cascatas de sinalização intracelular coordenadas pelas proteı́nas
da famı́lia MAPK, estando funcionalmente associado à regulação da plasticidade sináptica. A
interação do EGR-1 com o chumbo aumenta a ligação deste fator de transcrição ao DNA,
causando picos de ligação prematuros em algumas áreas do cérebro [90].
Além disso, ainda existem relatos que sugerem o papel do chumbo como agente de
interferência em canais iônicos dependentes de voltagem do tipo P, nos receptores nicotı́nicos
de acetilcolina e em outras proteı́nas relacionadas à plasticidade neuronal e ao desenvolvimento
neurológico [71, 82].
Efeitos tardios da intoxicação por chumbo

No Time-Interval 2 os clusteres, de uma maneira geral, sofreram um aumento no número de
nós e de conexões, passando a englobar 89,12% de todo o transcriptograma analisado. O número
de termos enriquecidos em cada cluster aumentou significativamente, tendo o seu número
total dobrado, havendo evidente conexão entre eles quando observados à luz do dendrograma
de cada cluster. A variedade de sistemas modulados, tanto positiva quanto negativamente,
também aumentou, passando a envolver uma gama enorme de processos biológicos. De forma
geral, todos os clusteres foram modulados positivamente quando comparados a seus correlatos
do Time-Interval 1.
Talvez a única grande exceção aqui seja o cluster E, pequeno (1,19% do total de nós)
e inexistente no T 1, que enriqueceu apenas 9 termos curiosamente correlacionados à defesa
e resposta imune a bactérias, fungos e outros agentes biológicos externos. É possı́vel que
Discussão 51
este padrão esteja relacionado a uma resposta autoimune, já que estudos em camundongos
mostraram que a exposição ao chumbo pode levar a este tipo de disfunção, com a produção
de anticorpos contra proteı́nas neurais, incluindo-se aı́ a proteı́na básica de mielina MBP e a
proteı́na glial GFAP, o que conduziria a quadros de doenças degenerativas do CNS induzidas
por chumbo [91]. Pode, também, ser a caracterização dos processos inflamatórios no tecido
cerebral descritos em [72] e não observados no T 1, manifestando-se aqui de forma tardia.
Outros distúrbios relacionados ao sistema imune, como imunossupressão e hipersensibilidade a
antı́genos, também foram relatados como estando associados à intoxicação pelo metal pesado
[92].
Esta alteração massiva observada tardiamente em todo o interatoma reforça a ideia na qual
o chumbo possui vocação para corromper de forma significativa o funcionamento de inúmeros
sistemas biológicos a ele expostos, substituindo ı́ons essenciais e alterando toda a homeostase
da transcrição. É assim, razoável supor que o desenvolvimento normal das células neuronais e
consequente funcionamento do CNS fique severamente comprometido como resultado de uma
exposição precoce e prolongada ao chumbo.
Efeitos fenotı́picos associados

A exposição ao chumbo durante o desenvolvimento do CNS, mesmo que a nı́veis considerados
baixos, pode afetar negativamente uma variedade de funções cognitivas e comportamentais,
resultando em deficit na aprendizagem, memória, atenção e função executiva de crianças,
cujas sequelas persistirão até a idade adulta. Apesar de já serem bem conhecidos os efeitos
diretos do P b2+ sobre a expressão gênica, seja por intermédio do estresse oxidativo, seja
pela sua capacidade em mimetizar ı́ons como cálcio e zinco, recentes estudos sugerem que
outros mecanismos podem agir como coadjuvantes. Dentre eles, a metilação do DNA pode ter
importância particular na explicação dos efeitos duradouros deste tipo de intoxicação.
De uma maneira geral, o aumento da metilação do DNA pode ser inversamente associado
aos valores de expressão gênica, cujos efeitos, em muitos casos, somente serão observados
anos mais tarde [93]. Deste modo, os efeitos persistentes da exposição precoce ao chumbo,
de forma não plenamente compreendida, podem envolver mecanismos epigenéticos, já que
são consistentes com um modelo de reprogramação fisiológica de base fetal que irão resultar
em uma incapacitação na vida adulta, especialmente quando ocorrem em sı́tios de proteı́nas
fundamentais à regulação transcricional [94].
Evidencias apontam para uma ação tardia de uma exposição pré-natal ao chumbo, man-
ifesta anos após o desaparecimento da intoxicação, relacionadas a problemas de atenção,
comportamentos de internalização (isolamento, queixas somáticas, ansiedade e depressão) e
externalização (quebrar de regras e comportamento agressivo), além de delinquência juvenil e
comportamento criminoso [95]. Estudos recentes também sugerem que alterações epigenéticas
causadas pelo chumbo no inı́cio da vida podem resultar em efeitos cardiovasculares adversos na
Discussão 52
vida adulta, como quadros de hipertensão, efeitos antifibrinolı́ticos e ativação pró-coagulante

de eritrócitos, dentre outros [96].
Ainda mais preocupantes são os relatos onde a herdabilidade de tais modificações epi-
genéticas resultariam em modificação de caracterı́sticas fenotı́picas de descendentes não expostos
ao P b2+ . Estudos sugerem que a exposição de mães ao chumbo durante a gravidez pode
alterar o status de metilação do DNA de células germinativas fetais dos filhos, detectáveis
em testes realizados no DNA dos netos [97] e que concentrações maternas de chumbo em
perı́odos muito anteriores à gravidez poderiam estar associadas ao aumento da pressão arterial
em seus filhos aos 10 anos de idade [96].
Paralelamente, muitos destes efeitos também podem estar associados a processos de
neurotransmissão. Diversos estudos demonstram que a exposição ao P b2+ tem potencial
para alterar a liberação de neurotransmissores, tanto em modelos in vivo quanto in vitro,
em vias glutamatérgicas e GABAérgicas, prejudicar o funcionamento de receptores NMDA,
causar alterações na transcrição do BDNF, no crescimento axonal e de neurites, modificar a
dinâmica de liberação vesicular e nı́veis de produção de neurotransmissores, além de causar
uma diminuição no número de terminais pré-sinápticos contendo múltiplas mitocôndrias. Em
especial, este último sugere que a disponibilidade de energia na forma de ATP também pode
ser comprometida, alterando, indiretamente, o mecanismo de liberação vesicular [98].
Desta forma nota-se que nossos achados também encontram-se em consonância com a
manifestação fenotı́pica das diversas sequelas causadas pelos efeitos sistêmicos da intoxicação
por chumbo, onde os inúmeros distúrbios, como aumento na agressividade, deficit de atenção,
prejuı́zos no desenvolvimento cerebral, aprendizado, memória e alterações cardiovasculares
podem ser arrolados como consequências diretas e mensuráveis.
53
Conclusão
O chumbo é um metal extremamente tóxico que atua de inúmeras formas no organismo.

Seus sintomas variados e amplamente conhecidos são o primeiro indı́cio de sua ação sistêmica.
Ao examinar sua atuação sobre componentes celulares, observa-se a ação em diversas partes da
célula, reflexo da habilidade deste metal em mimetizar ı́ons metálicos essenciais ao metabolismo
celular, alterando assim a conformação de metaloproteı́nas fundamentais a processos de sina-
lização responsáveis pelo acionamento de uma grande variedade de vias metabólicas importantes.
Sua interferência nas mais diversas camadas de regulação provoca efeitos generalizados e
severos em todo mecanismo de produção de metabólitos essenciais. Tais alterações são ainda
mais relevantes quando se trata do metabolismo celular neuronal. O comprometimento da
homeostase transcricional de vias relacionadas a modulação de microtúbulos, especificação
e crescimento axonal, liberação de neurotransmissores, excitabilidade neuronal, plasticidade
sináptica e potenciação de longa duração influenciam diretamente na capacidade do neurônio
interagir com neurônios vizinhos, com reflexos sobre as funções cognitivas, de memória e
capacidade de aprendizado compatı́veis com os sintomas descritos na literatura. Uma análise
baseada em biologia de sistemas mostrou-se fundamental para revelar, em um momento
inicial, a depleção de sistemas celulares vitais com possı́vel comprometimento do processo de
diferenciação celular, bem como o inı́cio de um possı́vel processo de modulação que acabou
por resultar, num segundo momento, em uma alteração massiva de grande parte dos sistemas
biológicos observados, com um possı́vel comprometimento de toda a função neuronal. Porém,
cabe aqui observar que todo este padrão relatado foi resultado de alterações transcricionais de
células neuronais em diferenciação expostas ao chumbo in vitro. Desta forma, estudos similares
se fazem necessários para confirmar a exatidão desta dinâmica em organismos vivos.
54
Referências
1 YAHALOM-MACK, N. et al. The earliest lead object in the levant. PLOS ONE, Public
Library of Science, v. 10, n. 12, p. 1–14, 12 2015.
2 KING, M. et al. Lead and lead alloys. In: . Kirk-Othmer Encyclopedia of Chemical
Technology. [S.l.]: American Cancer Society, 2014. p. 1–55. ISBN 9780471238966.
3 CARR, D. S. et al. Lead compounds. In: . Kirk-Othmer Encyclopedia of Chemical

Technology. [S.l.]: American Cancer Society, 2004. ISBN 9780471238966.
4 UNEP. Final review of scientific information on lead - Version of December

2010. 2010. Disponı́vel em: hhttps://www.cms.int/sites/default/files/document/
UNEP GC26 INF 11 Add 1 Final UNEP Lead review and apppendix Dec 2010.pdfi.
5 SANDERS, T. et al. Neurotoxic effects and biomarkers of lead exposure: a review. Rev
Environ Health, v. 24, n. 1, p. 15–45, 2009.
6 ORGANIZATION, W. H. Operational framework – Global Alliance to Eliminate Lead Paint.

2011. Disponı́vel em: hhttps://www.who.int/ipcs/assessment/public health/gaelp/en/i.
7 NATIONAL Surveillance Data Table. US Centers for Disease Control and

Prevention (CDC), 2019. Disponı́vel em: hhttps://www.cdc.gov/nceh/lead/docs/
CBLS-National-Table-Update-042619.xlsxi.
8 ACCLP. Low Level Lead Exposure Harms Children: A Renewed Call for Primary Prevention.
Atlanta, GA, 2012. 1–54 p. Disponı́vel em: hhttp://www.cdc.gov/nceh/lead/acclpp/
final document 030712.pdfi.
9 PORTARIA/GM/MS no 1339, de 18 de novembro de 1999. 1999.
10 Resolução no 257 de 30 de junho de 1999. 1999.
11 Resolução RDC no 306 de 7 de dezembro de 2004. 2004.
12 LOPES, A. C. B. A. et al. Association between blood lead and blood pressure: a

population-based study in Brazilian adults. Environ Health, v. 16, n. 1, p. 27, 03 2017.
13 PAOLIELLO, M. M.; CAPITANI, E. M. D. Environmental contamination and human

exposure to lead in Brazil. Rev Environ Contam Toxicol, v. 184, p. 59–96, 2005.
14 AMARAL, V. S. do et al. Environmental impact of natural lead occurrence on a population

settled in the Brazilian North East Region. Toxicology Letters, 2016.
15 CAVALLERI, A. et al. Determination of plasma lead levels in normal subjects and in

lead-exposed workers. Br J Ind Med, v. 35, n. 1, p. 21–26, Feb 1978.
16 SAKAI, T. Biomarkers of lead exposure. Ind Health, v. 38, n. 2, p. 127–142, Apr 2000.
17 MCLACHLIN, J. R.; GOYER, R. A.; CHERIAN, M. G. Formation of lead-induced inclusion

bodies in primary rat kidney epithelial cell cultures: effect of actinomycin D and cycloheximide.
Toxicol. Appl. Pharmacol., v. 56, n. 3, p. 418–431, Dec 1980.
Referências 55
18 SHELTON, K. R. et al. Low-abundance 32-kilodalton nuclear protein specifically enriched

in the central nervous system. J. Neurosci. Res., v. 25, n. 3, p. 287–294, Mar 1990.
19 ZUO, P. et al. Potential role of alpha-synuclein and metallothionein in lead-induced

inclusion body formation. Toxicol. Sci., v. 111, n. 1, p. 100–108, Sep 2009.
20 GONICK, H. C. Lead-binding proteins: a review. J Toxicol, v. 2011, p. 686050, 2011.
21 VIGEH, M.; SMITH, D. R.; HSU, P. C. How does lead induce male infertility? Iran J
Reprod Med, v. 9, n. 1, p. 1–8, 2011.
22 ZHANG, H. et al. Binding mode investigations on the interaction of lead(II) acetate with
human chorionic gonadotropin. J Phys Chem B, v. 118, n. 32, p. 9644–9650, Aug 2014.
23 SOUZA, I. D. de; ANDRADE, A. S. de; DALMOLIN, R. J. S. Lead-interacting proteins

and their implication in lead poisoning. Critical Reviews in Toxicology, Informa UK Limited,
trading as Taylor & Francis Group, v. 0, n. 0, p. 1–12, 2018. ISSN 1040-8444.
24 BARANOWSKA-BOSIACKA, I. et al. Neurotoxicity of lead. Hypothetical molecular

mechanisms of synaptic function disorders. Neurologia I Neurochirurgia Polska, v. 46, p.
569–578, 2012. ISSN 00283843.
25 NIGG, J. T. ADHD, lead exposure and prevention: how much lead or how much evidence
is needed? Expert Rev Neurother, v. 8, n. 4, p. 519–521, Apr 2008.
26 BELLINGER, D. C.; NEEDLEMAN, H. L. Intellectual Impairment and Blood Lead Levels.

New England Journal of Medicine, v. 349, n. 5, p. 500–502, jul 2003. ISSN 0028-4793.
27 PORTBURY, S. D.; ADLARD, P. A. Zinc Signal in Brain Diseases. Int J Mol Sci, v. 18,
n. 12, Nov 2017.
28 CASSANDRI, M. et al. Zinc-finger proteins in health and disease. Cell Death Discov, v. 3,
p. 17071, 2017.
29 MURN, J. et al. Control of a neuronal morphology program by an RNA-binding zinc finger

protein, Unkempt. Genes Dev., v. 29, n. 5, p. 501–512, Mar 2015.
30 NOH, S. et al. The direct modulatory activity of zinc toward ion channels. Integr Med
Res, v. 4, n. 3, p. 142–146, Sep 2015.
31 SUN, H. S. et al. Zn2+ sensitivity of high- and low-voltage activated calcium channels.
Biophys. J., v. 93, n. 4, p. 1175–1183, Aug 2007.
32 JOHNSON, C. K. Calmodulin, conformational states, and calcium signaling. A

single-molecule perspective. Biochemistry, v. 45, n. 48, p. 14233–14246, Dec 2006.
33 SOLA, C. et al. The Ca2+/calmodulin system in neuronal hyperexcitability. Int. J.

Biochem. Cell Biol., v. 33, n. 5, p. 439–455, May 2001.
34 CALLENDER, J. A.; NEWTON, A. C. Conventional protein kinase c in the brain: 40

years later. Neuronal Signaling, Portland Press Journals portal, v. 1, n. 2, 2017.
35 WAYMAN, G. A. et al. Calmodulin-kinases: modulators of neuronal development and

plasticity. Neuron, v. 59, n. 6, p. 914–931, Sep 2008.
Referências 56
36 MITALIPOV, S.; WOLF, D. Totipotency, pluripotency and nuclear reprogramming. In:

. Engineering of Stem Cells. Berlin, Heidelberg: Springer Berlin Heidelberg, 2009. p.
185–199. ISBN 978-3-540-88806-2.
37 MARTINEZ-CERDENO, V.; NOCTOR, S. C. Neural Progenitor Cell Terminology. Front

Neuroanat, v. 12, p. 104, 2018.
38 TANG, Y.; YU, P.; CHENG, L. Current progress in the derivation and therapeutic
application of neural stem cells. Cell Death Dis, v. 8, n. 10, p. e3108, 10 2017.
39 KANEKO, Y. et al. Musashi1: an evolutionally conserved marker for CNS progenitor cells
including neural stem cells. Dev. Neurosci., v. 22, n. 1-2, p. 139–153, 2000.
40 WIESE, C. et al. Nestin expression–a property of multi-lineage progenitor cells? Cell. Mol.
Life Sci., v. 61, n. 19-20, p. 2510–2522, Oct 2004.
41 LUTOLF, S. et al. Notch1 is required for neuronal and glial differentiation in the
cerebellum. Development, v. 129, n. 2, p. 373–385, Jan 2002. ISSN 1477-9129.
42 SUTER, D. M. et al. A Sox1 to Pax6 switch drives neuroectoderm to radial glia

progression during differentiation of mouse embryonic stem cells. Stem Cells, v. 27, n. 1, p.
49–58, Jan 2009.
43 WHITE, R. B.; THOMAS, M. G. Moving beyond tyrosine hydroxylase to define

dopaminergic neurons for use in cell replacement therapies for Parkinson’s disease. CNS Neurol
Disord Drug Targets, v. 11, n. 4, p. 340–349, Jun 2012.
44 KHAN, S. et al. Survival of a Novel Subset of Midbrain Dopaminergic Neurons Projecting

to the Lateral Septum Is Dependent on NeuroD Proteins. J. Neurosci., v. 37, n. 9, p.
2305–2316, 03 2017.
45 RAO, M. S.; SHETTY, A. K. Efficacy of doublecortin as a marker to analyse the absolute

number and dendritic growth of newly generated neurons in the adult dentate gyrus. Eur. J.
Neurosci., v. 19, n. 2, p. 234–246, Jan 2004.
46 MATHEWS, K. J. et al. Evidence for reduced neurogenesis in the aging human

hippocampus despite stable stem cell markers. Aging Cell, v. 16, n. 5, p. 1195–1199, 10 2017.
47 GUSEL’NIKOVA, V. V.; KORZHEVSKIY, D. E. NeuN As a Neuronal Nuclear Antigen and

Neuron Differentiation Marker. Acta Naturae, v. 7, n. 2, p. 42–47, 2015.
48 COSSETTE, M.; LEVESQUE, D.; PARENT, A. Neurochemical characterization of

dopaminergic neurons in human striatum. Parkinsonism Relat. Disord., v. 11, n. 5, p. 277–286,
Aug 2005.
49 MORAIS, D. A. A.; ALMEIDA, R. M. C.; DALMOLIN, R. J. S. Transcriptogramer: an

R/Bioconductor package for transcriptional analysis based on protein-protein interaction.
Bioinformatics (Oxford, England), p. 1–2, 01 2019.
50 RYBARCZYK-FILHO, J. L. et al. Towards a genome-wide transcriptogram: the

saccharomyces cerevisiae case. Nucleic Acids Res, Oxford University Press, v. 39, n. 8, p.
3005–3016, Apr 2011. ISSN 0305-1048. Gkq1269[PII].
Referências 57
51 RITCHIE, M. E. et al. limma powers differential expression analyses for rna-sequencing

and microarray studies. Nucleic Acids Research, v. 43, n. 7, p. e47, 2015.
52 ALEXA, A.; RAHNENFUHRER, J. topGO: Enrichment Analysis for Gene Ontology. [S.l.],
2016. R package version 2.32.0.
53 LIDSKY, T. I.; SCHNEIDER, J. S. Lead neurotoxicity in children: basic mechanisms and

clinical correlates. Brain, v. 126, n. Pt 1, p. 5–19, Jan 2003.
54 WHITE, L. D. et al. New and evolving concepts in the neurotoxicology of lead. Toxicol.
Appl. Pharmacol., v. 225, n. 1, p. 1–27, Nov 2007.
55 NATIONAL Surveillance Data Table. US Centers for Disease Control and

Prevention (CDC), 2016. Disponı́vel em: hhttps://wwwn.cdc.gov/nndss/conditions/
lead-elevated-blood-levels-children/case-definition/2016/i.
56 SCHNUR, J.; JOHN, R. M. Childhood lead poisoning and the new centers for disease
control and prevention guidelines for lead exposure. Journal of the American Association of
Nurse Practitioners, v. 26, n. 5, p. 238–247, 2014. ISSN 23276924.
57 MEYER, P. A.; BROWN, M. J.; FALK, H. Global approach to reducing lead exposure and
poisoning. Mutation Research/Reviews in Mutation Research, v. 659, n. 1, p. 166–175, 2008.
ISSN 13835742.
58 SCHNEIDER, J. S.; METTIL, W.; ANDERSON, D. W. Differential effect of postnatal

lead exposure on gene expression in the hippocampus and frontal cortex. J Mol Neurosci,
v. 47, n. 1, p. 76–88, 2012. ISSN 0895-8696.
59 MITRA, P. et al. Clinical and molecular aspects of lead toxicity: An update. Critical
Reviews in Clinical Laboratory Sciences, Informa Healthcare USA, Inc, v. 54, n. 7-8, p.
506–528, 2017. ISSN 1549781X.
60 SENUT, M.-C. et al. Lead Exposure Disrupts Global DNA Methylation in Human
Embryonic Stem Cells and Alters Their Neuronal Differentiation. Toxicological Sciences, v. 139,
n. 1, p. 142–161, may 2014. ISSN 1096-6080.
61 ABDULLAH, M. et al. Diverse effects of lead nitrate on the proliferation, differentiation,

and gene expression of stem cells isolated from a dental origin. TheScientificWorldJournal,
Hindawi, v. 2014, p. 1–12, jan 2014. ISSN 1537-744X.
62 KAGEYAMA, R. et al. Dynamic regulation of notch signaling in neural progenitor cells.

Current Opinion in Cell Biology, v. 21, n. 6, p. 733 – 740, 2009. ISSN 0955-0674. Cell
differentiation / Cell division, growth and death.
63 CRUMPTON, T. et al. Lead exposure in pheochromocytoma (PC12) cells alters neural

differentiation and Sp1 DNA-binding. NeuroToxicology, v. 22, n. 1, p. 49–62, 2001. ISSN
0161813X.
64 HUANG, F.; SCHNEIDER, J. Effects of Lead Exposure on Proliferation and Differentiation

of Neural Stem Cells Derived from Different Regions of Embryonic Rat Brain. NeuroToxicology,
Elsevier, v. 25, n. 6, p. 1001–1012, dec 2004. ISSN 0161-813X.
Referências 58
65 ORDEMANN, J. M.; AUSTIN, R. N. Lead neurotoxicity: exploring the potential impact

of lead substitution in zinc-finger proteins on mental health. Metallomics, Royal Society of
Chemistry, v. 8, n. 6, p. 579–88, 2016. ISSN 1756-5901.
66 LISMAN, J.; YASUDA, R.; RAGHAVACHARI, S. Mechanisms of CaMKII action in

long-term potentiation. Nat. Rev. Neurosci., v. 13, n. 3, p. 169–182, Feb 2012.
67 XIA, Z.; STORM, D. R. The role of calmodulin as a signal integrator for synaptic
plasticity. Nature Reviews Neuroscience, Nature Publishing Group, v. 6, n. 4, p. 267–276, apr
2005. ISSN 1471-003X.
68 HABERMANN, E.; CROWELL, K.; JANICKI, P. Lead and other metals can substitute for
ca2+ in calmodulin. Archives of Toxicology, v. 54, n. 1, p. 61–70, Sep 1983. ISSN 1432-0738.
69 SUN, X. et al. Analysis of differential effects of Pb2+ on protein kinase C isozymes.

Toxicol. Appl. Pharmacol., v. 156, n. 1, p. 40–45, Apr 1999.
70 BOUTON, C. M. L. S. et al. Synaptotagmin I is a molecular target for lead. Journal of

Neurochemistry, v. 76, n. 6, p. 1724–1735, 2001. ISSN 00223042.
71 KASTEN-JOLLY, J.; HEO, Y.; LAWRENCE, D. A. Central nervous system cytokine gene
expression: Modulation by lead. Journal of Biochemical and Molecular Toxicology, v. 25, n. 1,
p. 41–54, 2011. ISSN 10956670.
72 KASTEN-JOLLY, J. et al. Developmental lead effects on behavior and brain gene

expression in male and female BALB/cAnNTac mice. Neurotoxicology, v. 33, n. 5, p.
1005–1020, Oct 2012.
73 NIHEI, M. K.; GUILARTE, T. R. NMDAR-2A subunit protein expression is reduced in the

hippocampus of rats exposed to Pb2+ during development. Brain Res. Mol. Brain Res., v. 66,
n. 1-2, p. 42–49, Mar 1999.
74 NIHEI, M. K.; GUILARTE, T. R. Molecular changes in glutamatergic synapses induced by

Pb2+: association with deficits of LTP and spatial learning. Neurotoxicology, v. 22, n. 5, p.
635–643, Oct 2001.
75 HARTWIG, A. Role of DNA repair inhibition in lead- and cadmium-induced genotoxicity:

a review. Environ. Health Perspect., v. 102 Suppl 3, p. 45–50, Sep 1994.
76 JANNUZZI, A. T.; ALPERTUNGA, B. Evaluation of DNA damage and DNA repair

capacity in occupationally lead-exposed workers. Toxicol Ind Health, v. 32, n. 11, p. 1859–1865,
Nov 2016.
77 BOCKAERT, J.; MARIN, P. mtor in brain physiology and pathologies. Physiological

Reviews, v. 95, n. 4, p. 1157–1187, 2015. PMID: 26269525.
78 THOMANETZ, V. et al. Ablation of the mTORC2 component rictor in brain or Purkinje

cells affects size and neuron morphology. J. Cell Biol., v. 201, n. 2, p. 293–308, Apr 2013.
79 LIPTON, J. O.; SAHIN, M. The neurology of mTOR. Neuron, v. 84, n. 2, p. 275–291,

Oct 2014.
80 SWITON, K. et al. Molecular neurobiology of mTOR. Neuroscience, v. 341, p. 112–153,

01 2017.
Referências 59
81 NEAL, A. P. et al. Lead exposure during synaptogenesis alters vesicular proteins and
impairs vesicular release: potential role of NMDA receptor-dependent BDNF signaling. Toxicol.
Sci., v. 116, n. 1, p. 249–263, Jul 2010.
82 STANSFIELD, K. H. et al. Dysregulation of BDNF-TrkB signaling in developing

hippocampal neurons by Pb 2+: Implications for an environmental basis of neurodevelopmental
disorders. Toxicological Sciences, v. 127, n. 1, p. 277–295, 2012. ISSN 10966080.
83 TOSCANO, C. D.; GUILARTE, T. R. Lead neurotoxicity: From exposure to molecular

effects. Brain Research Reviews, v. 49, n. 3, p. 529–554, 2005. ISSN 01650173.
84 GUILARTE, T. R.; OPLER, M.; PLETNIKOV, M. Is lead exposure in early life an

environmental risk factor for Schizophrenia? Neurobiological connections and testable
hypotheses. Neurotoxicology, v. 33, n. 3, p. 560–574, Jun 2012.
85 MELDRUM, B. S. Glutamate as a neurotransmitter in the brain: review of physiology and

pathology. J. Nutr., v. 130, n. 4S Suppl, p. 1007S–15S, 04 2000.
86 NEWCOMER, J. W.; FARBER, N. B.; OLNEY, J. W. NMDA receptor function, memory,

and brain aging. Dialogues Clin Neurosci, v. 2, n. 3, p. 219–232, Sep 2000.
87 PAOLETTI, P.; BELLONE, C.; ZHOU, Q. NMDA receptor subunit diversity: impact
on receptor properties, synaptic plasticity and disease. Nat. Rev. Neurosci., v. 14, n. 6, p.
383–400, Jun 2013.
88 GUILARTE, T. R.; MCGLOTHAN, J. L. Hippocampal NMDA receptor mRNA undergoes

subunit specific changes during developmental lead exposure. Brain Research, v. 790, n. 1, p.
98–107, 1998. ISSN 00068993.
89 NIHEI, M. K. et al. N-methyl-D-aspartate receptor subunit changes are associated with

lead-induced deficits of long-term potentiation and spatial learning. Neuroscience, v. 99, n. 2,
p. 233–242, 2000. ISSN 03064522.
90 REDDY, G. R.; ZAWIA, N. H. Lead exposure alters Egr-1 DNA-binding in the neonatal
rat brain. Int. J. Dev. Neurosci., v. 18, n. 8, p. 791–795, Dec 2000.
91 WATERMAN, S. J.; FAWAL, H. A. el; SNYDER, C. A. Lead alters the immunogenicity of

two neural proteins: a potential mechanism for the progression of lead-induced neurotoxicity.
Environmental Health Perspectives, v. 102, n. 12, p. 1052–1056, 1994.
92 MISHRA, K. Lead exposure and its impact on immune system: A review. Toxicology in
Vitro, v. 23, n. 6, p. 969 – 972, 2009. ISSN 0887-2333.
93 WRIGHT, R. O. et al. Biomarkers of lead exposure and DNA methylation within

retrotransposons. Environ. Health Perspect., v. 118, n. 6, p. 790–795, Jun 2010.
94 SCHNEIDER, J. S.; KIDD, S. K.; ANDERSON, D. W. Influence of developmental lead

exposure on expression of DNA methyltransferases and methyl cytosine-binding proteins in
hippocampus. Toxicol. Lett., v. 217, n. 1, p. 75–81, Feb 2013.
95 LIU, J.; ZHAO, S. R.; REYES, T. Neurological and Epigenetic Implications of Nutritional
Deficiencies on Psychopathology: Conceptualization and Review of Evidence. Int J Mol Sci,
v. 16, n. 8, p. 18129–18148, Aug 2015.
Referências 60
96 ENGSTRöM, K. et al. Prenatal lead exposure is associated with decreased cord blood
DNA methylation of the glycoprotein VI gene involved in platelet activation and thrombus
formation. Environmental Epigenetics, v. 1, n. 1, 11 2015. ISSN 2058-5888. Dvv007. Disponı́vel
em: hhttps://doi.org/10.1093/eep/dvv007i.
97 SEN, A. et al. Multigenerational epigenetic inheritance in humans: DNA methylation

changes associated with maternal exposure to lead can be transmitted to the grandchildren.
Sci Rep, v. 5, p. 14466, Sep 2015.
98 ZHANG, X. L. et al. Presynaptic mechanisms of lead neurotoxicity: effects on vesicular

release, vesicle clustering and mitochondria number. PLoS ONE, v. 10, n. 5, p. e0127461,
2015.
61
APÊNDICE A – Material suplementar do

artigo
Supplementary Material
SUPPLEMENTARY FIGURES
7 ●
4 ●
5 ● 8 ● 9 ●
11 ●
10 ● 13 ●
6 ●
50
12 ● ●
14 ●
3 ●
15 19 ●
●
16 ●
21 ●
25 ●
2 ●
18 ●
1 1 ●
0
2
26 ● 20 ● ● ●
4 23
3
7
0
24 ●
9
5 11 13
17
PC2
25
6
19
8
15
−50
10 14
12 26
18
22 23
20
Control
16
● Lead 30
−100
24
−150 −100 −50 0 50

PC1
Figure S1: PCA after expression normalization. PC1xPC2. Blue dots designate the control, and red dots
the 30µM samples. The number indicates the time of cell exposition to lead.
1
100
75
Percentage of explained variances
50
Cumulative
25
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
Dimensions
Figure S2: Percentage of explained variances of Lead30 samples used for clusterization. Blue bars indicate
the explained variances of the first 17 components and the orange line it cumulative total (95.372%).
2
7
4 5 8
50 6
9 10
11
12
●
●
3 13 15
●
●
14
17 ●
0 ●
16
● ● ● 18
19 ● 21
20
●
22
PC2
● ●
2 25 ● 23
26 ●
−50 ● 24
cluster
−100
●
a● 1
a 2
a 3
1
−150
−200 −100 0
PC1
Figure S3: PCA clusterization of the Lead30 samples. Colors designate the different clusters found using
the R package FactoMineR’s HCPC function whit autodetection of the number of clusters (k).
Frontiers 3
MSI1 NES
1.0
0.9
0.8
Expression (log-CPM)
0.6
Lead30 0.5 Lead30
Control Control
0.3
p−val: 7.40E−04 p−val: 2.58E−08 0.2 p−val: 8.23E−05 p−val: 1.84E−03
Time−interval 1 Time−interval 2 Time−interval 1 Time−interval 2

0.0 0.0
0 10 20 0 10 20
Days Days
NOTCH1 SOX1
1.0
0.9
0.8
0.6
Lead30 0.5 Lead30
Control Control
0.3
p−val: 4.11E−05 p−val: 2.58E−08 0.2 p−val: 8.23E−05 p−val: 7.66E−03

0.0 0.0
0 10 20 0 10 20
Days Days
Figure S4: NPC cells markers progression time-line. Graphics show the time points of control group
expression values for NPC cells markers (dotted lines), and lead-treated samples expression values (solid
lines). Colors represent distinct markers. Numbers at X-axis identify the days of treatment. Numbers
at Y-axis represent the expression values of samples, in log-CPM. Shaded areas inside graphical area
delimit the time-intervals 1 and 2, from day 3 to 11, and day 12 to 26, respectively. Kolmogorov-Smirnov
test comparing the entire timeline of lead-treated and control samples determine significant differences
between datasets with p-values lower than 6 × 10−5 for all markers. Kolmogorov-Smirnov test p-values of
comparison between lead-treated and control samples in time-interval 1 and time-interval 2, when taken in
isolation, have it p-values corrected by FDR annotated inside the graphic. Blue p-values indicate significant
differences between distributions (p-value ≤ 0.01).
4
DCX GAD2
1.5 12.0
1.0 8.0
Lead30 Lead30
Control Control
0.5 4.0
p−val: 8.23E−05 p−val: 2.62E−02 p−val: 3.52E−01 p−val: 7.74E−07

0.0 0.0
0 10 20 0 10 20
Days Days
GAD1 NEUROD6
3.0
4.0
3.0
2.0
Lead30 Lead30
2.0
Control Control
1.0
1.0 p−val: 3.36E−02 p−val: 2.58E−08 p−val: 7.30E−01 p−val: 7.74E−07

0.0 0.0
0 10 20 0 10 20
Days Days
RBFOX3 TH
Expression (log-CPM) 4.0
4.0
3.0
Lead30 Lead30
2.0
2.0 Control Control
p−val: 4.11E−05 p−val: 2.58E−08 p−val: 1.26E−01 p−val: 1.05E−04
1.0
0.0 0.0
0 10 20 0 10 20
Days Days
Figure S5: Neuronal cells markers progression time-line. Graphics show the time points of control group
expression values for Neuronal cells markers (dotted lines), and lead-treated samples expression values
(solid lines). Colors represent distinct markers. Numbers at X-axis identify the days of treatment. Numbers
at Y-axis represent the expression values of samples, in log-CPM. Shaded areas inside graphical area
delimit the time-intervals 1 and 2, from day 3 to 11, and day 12 to 26, respectively. Kolmogorov-Smirnov
test comparing the entire timeline of lead-treated and control samples determine significant differences
between datasets with p-values lower than 6 × 10−5 for all markers. Kolmogorov-Smirnov test p-values
of comparison between lead-treated and control samples in time-interval 1 and time-interval 2, when
taken in isolation, have it p-values corrected by FDR annotated inside the graphic. Red p-values indicate
non-significant differences between distributions and blue p-values indicate significant ones (p-value
≤ 0.01).
Frontiers 5
0.65
0.55
0.45
Difference of means (case − control)

0.35
0.25
0.15
0.05
−0.05
−0.15
−0.25
−0.35
−0.45
−0.55
−0.65
0 1000 2000 3000 4000 5000 6000 7000 8000 9000 10000 11000 12000 13000 14000 15000
Gene position
cell communication
ER to Golgi vesicle-mediated transport
cell activation
positive regulation of macromolecule met...

cellular response to extracellular stimu...
peptidyl-serine modification
response to virus signalin...

regulation of cytokine-mediated
immune system process cellular response to external stimulus
vesicle organization positive regulation of RNA biosynthetic ...
cell death regulation of response to cytokine stimu...
Cluster 5
transmembrane receptor protein serine/th... innate immune response positive regulation of cellular biosynth...
signaling cell-cell signaling by wnt positive regulation of nucleic acid-temp...
regulation of cell death carbohydrate homeostasis positive regulation of biosynthetic proc...

positive regulation of cellular protein ...
I-kappaB
positive regulation of transcription by ... kinase/NF-kappaB signaling
defense response to virus positive regulation of gene expression
Golgi vesicle transport maintenance of cell number
vesicle-mediated transport regulation of cysteine-type endopeptidas...
positive regulation of transcription, DN... positive regulation of RNA metabolic pro...
positive regulation of nitrogen compound... regulation of DNA binding transcription ...
regulation of I-kappaB kinase/NF-kappaB ... positive regulation of nucleobase-contai...
secretion by cell regulation of innate immune response
negative regulation of cell differentiat... positive regulation of macromolecule bio...

COPII vesicle coating
Cluster 6
cardiac septum development positive regulation of DNA bindingresponse
trans... to biotic stimulus
organelle localization regulation of cellular macromolecule bio...
transcription, DNA-templated
cellular nitrogen compound biosynthetic ...

activation of innate immune response
establishment of vesicle localization
29 terms
cellular response to organic cyclic comp... aromatic compound biosynthetic process
vesicle budding from membrane localization
intracellular transport response to steroid hormone organic cyclic compound biosynthetic pro... regulation of transcription, DNA-templat...
membrane organization
vesicle targeting
establishment of localization in cell heart development nucleobase-containing compound biosynthe...
DNA-templated transcription, initiation RNA biosynthetic process
hormone-mediated signaling pathway
cellular localization neural tube development
establishment of organelle localization heterocycle biosynthetic process
negative regulation of cell death cellular response

intracellular receptor to steroid
signaling pathway hormone
cell sti...
fate commitment
cardiac chamber development
RNA biosynthetic process cellular catabolic process
catabolic process regulation of catabolic process
pattern specification process
regulation of cellular catabolic process
embryo development
heterocycle biosynthetic process process utilizing autophagic mechanism
embryonic morphogenesis
heart morphogenesis cellular macromolecule metabolic process
organic cyclic compound biosynthetic pro...
cell cycle G2/M phase transition autophagy
regulation of cellular process multicellular organismal process

aromatic compound biosynthetic process regulation of G2/M transition of mitotic... cellular protein modification process
membrane docking organonitrogen compound metabolic proces...

regulation of biological process
regulation of cell cycle protein modification process
negative regulation of cellular metaboli...
cellular localization
G2/M transition of mitotic cell cycle
generation of precursor metabolites and ... biological regulation macromolecule modification
regulation of cellregulation of mitotic
cycle G2/M phase cell cycle phase t...
tran... cellular process
positive
regulation of protein regulation
metabolic of metabolic process
process
response to stimulus negative regulation of metabolic process
oxidative phosphorylation cellular component assembly
cell communication
cellular response to stimulus
regulation of microtubule-based process

organelle localization
signal transduction
nucleoside triphosphate metabolic proces... microtubule
microtubule-based process cytoskeleton organization
protein metabolic process
cellular component biogenesis regulation of cellular protein metabolic...
cilium assembly signaling

positive regulation of cellular metaboli...
cell division organelle assembly cellular component organization
energy derivation by oxidation of organi... cilium organization
positive regulation of nitrogen compound...

cellular component organization or bioge...
cell cycle process cytoskeleton organization cell projection assembly small molecule biosynthetic process
electron transport small
chain molecule metabolic process cell cycle
regulation of lipid metabolic process localization
cell proliferation
plasma membrane bounded cell projection ...
tube developmentnegative regulation of developmental pro...
purine nucleotide metabolic process organelle organization positive regulation of biological proces...regulation of molecular function
positive regulation of cellular process cell surface receptor signaling pathway

drug metabolic process growth
regulation of small molecule metabolic p...
cellular respiration
Cluster 11
apoptotic signaling pathway
cell projection organization
oxidation-reduction process negative regulation of response to stimu...
negative regulation of multicellular org... positive regulation of response to stimu...
negative regulation of biological proces...
nucleoside phosphate metabolic process
organophosphate metabolic process negative regulation of cell communicatio...
regulation of response to stimulus
phosphorus metabolic process
ion transmembrane transport 188 terms negative regulation of signaling

regulation of nitrogen compound metaboli...
positive regulation of signaling

regulation of cell death
regulation of cellular component organiz...
negative regulation of cellular process
cell death
nucleoside phosphate
nucleotide catabolic
catabolic process
process positive regulation of cell communicatio...
nucleoside monophosphate metabolic proce...
positive regulation of cellular protein ...
regulation of metabolic process positive regulation of developmental pro... cellular response to chemical stimulus
purine nucleoside monophosphate biosynth...
transmembrane transport regulation of primary metabolic process
regulation of cell communication immune system process
positive regulation of signal transducti...
response to organic substance
carbohydrate derivative metabolic proces... nucleoside triphosphate
nucleosidebiosynthetic pro... biosynthetic pr...
monophosphate
regulation of multicellular organismal p...
ATP metabolic process regulation of phosphate metabolic proces...
response to endogenous stimulus
ATP generation from ADP
regulation of cellular metabolic process
positive regulation of protein metabolic...
regulation of macromolecule metabolic pr...
pyridine-containing compound metabolic p... regulation of developmental
regulation process
of signal transduction
20 terms ribose phosphate metabolic process

pyridine nucleotide metabolic process
glycolytic process
regulation of protein modification proce...
regulation of phosphorus metabolic proce...
protein phosphorylation
regulation of signaling
cellular response to organic substance
regulation of cell differentiation
nucleotide phosphorylation
purine-containing compound metabolic pro... developmental process regulation of response to stress
pyridine-containing compound biosyntheti...
purine ribonucleoside triphosphate biosy...
ADP metabolic process
ATP biosynthetic process positive regulation of multicellular org...
multi-organism process
pyridine nucleotide biosynthetic process
nucleobase-containing small molecule met...
pyruvate metabolic process
nucleoside diphosphate metabolic process cellular developmental process
movement of cell or subcellular monosaccharide
componen... metabolic process
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
Normalized term rate
Figure S6: Connectivity and GO terms of clusters 5, 6, and 11. Transcriptogram, cluster’s names, and
colors correspond to the time-interval 1. Top graph of each cluster represents the connectivity of PPI
networks. Nodes are the cluster’s relevant proteins but are not explicitly represented. The confluence of
edges identifies them. Botton graph of each cluster is a dendrogram and refers to the cluster’s enriched GOs
hierarchy. Circles represent each enriched GO term. Node sizes are proportional to the numbers of terms
held by each GO term. Dendrogram colors represent the normalized terms occupation rate, where dark
colors indicate a larger number of genes detected over all genes belonging to the GO. Distance between
two nodes is proportional to the number of genes they have in common.
6
Cluster B
Cluster 5
Cluster 6
Cluster 4
183 terms
245 terms
188 terms
0.65
54 terms
Cluster A 0.55
0.45
Difference of means (case − control) 0.35
0.25
123 terms
0.15
Cluster E
0.05
−0.05
−0.15
−0.25 9 terms
−0.35
A 4 B 5 6 7 8 C 11 D E A
−0.45
−0.55
−0.65
0 1000 2000 3000 4000 5000 6000 7000 8000 9000 10000 11000 12000 13000 14000 15000
Cluster D
Cluster 7 Gene position
27 terms
Cluster C
Cluster 8
Cluster 11
47 terms
264 terms
22 terms
108 terms
Figure S7: Connectivity and GO terms of time-interval 2. Transcriptogram, cluster’s names, and colors
correspond to the time-interval 2. Dendrograms refers to the cluster’s enriched GOs hierarchy. Its node
sizes are proportional to the numbers of terms held by each GO term. Dendrogram colors represent the
normalized terms occupation rate, where dark colors indicate a larger number of genes detected over all
genes belonging to the GO.
Frontiers 7
Figures S6 to S15 are dendrograms of clusters 1 to 11 of time-interval 1, respectively, and refers to the
cluster’s enriched GOs hierarchy. Circles represent each enriched GO term and are labeled by it respective
description. Node sizes are proportional to the numbers of terms held by each GO term. Dendrogram
colors represent the normalized terms occupation rate, where dark colors indicate a larger number of genes
detected over all genes belonging to the GO. Distance between two nodes is proportional to the number of
genes they have in common. Cluster 2 do not have a dendrogram because it had no GO terms enriched.
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
cellular nitrogen compound metabolic pro...
regulation of cellular biosynthetic proc...
nucleobase-containing compound biosynthe...
regulation of biosynthetic process
organic substance biosynthetic process

regulation of transcription, DNA-templat...
gene expressionregulation of macromolecule metabolic pr...
regulation of cellular macromolecule bio...

cellular biosynthetic process regulation of cellular metabolic process
regulation of macromolecule biosynthetic...
[4954.2, 5154.4)
regulation of RNA metabolic process heterocycle metabolic process
regulation of gene expression
cellular macromolecule biosynthetic proc... RNA biosynthetic process

biosynthetic process
Term size (37 terms)
nucleobase-containing compound metabolic... transcription by RNA polymerase II
aromatic compound biosynthetic process

[4511.2, 4807.8) regulation of primary metabolic process
macromolecule biosynthetic process

cellular macromolecule metabolic process heterocycle biosynthetic process
regulation of nucleobase-containing comp...
cellular aromatic compound metabolic pro...
RNA metabolic process transcription, DNA-templated
[3527.2, 3956)
nucleic acid metabolic process
regulation of metabolic process
macromolecule metabolic process
regulation of RNA biosynthetic process
[3209, 3388.2)
organic cyclic compound metabolic proces...
[2189, 2899.4)
Figure S8: Complete dendrogram of cluster 1 – time-interval 1
8
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
protein complex oligomerization
cellular potassium ion transport

ion transport
transmembrane transport
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
protein-containing complex remodeling
inositol phosphate metabolic process
biological adhesion
organic hydroxy compound metabolic proce...

regulation of protein activation cascade
protein-lipid complex subunit organizati...
response to wounding
regulation of small GTPase mediated sign...
anatomical structure morphogenesis

coagulation regulation of protein maturation
cell communication
organic hydroxy compound biosynthetic pr...

alcohol metabolic process
protein activation cascade
signaling
small GTPase mediated signal transductio...
regulation of hemostasis
extracellular structure organization
Frontiers 9
0.0

0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
intracellular transport
0.9
1.0
RNA biosynthetic process
vesicle targeting
vesicle budding from membrane
organic cyclic compound biosynthetic pro... membrane organization
aromatic compound biosynthetic process establishment of vesicle localization

[3109.8, 3870.8)
organelle localization
positive regulation of nitrogen compound...
negative regulation of cell death
heterocycle biosynthetic process
establishment of organelle localization Golgi vesicle transportsignaling
establishment of localization in cell

vesicle organization
[1831.2, 2540.8) cell communication
localization COPII vesicle coating
secretion by cell
vesicle-mediated transport ER to Golgi vesicle-mediated transport

regulation of cell death
[1167, 1531) cell death

cell activation
immune system process
[303.2, 606.2)
[63, 169)
0.0

0.1
0.2
0.3
positive regulation of cellular biosynth... positive regulation of metabolic process 0.4
positive regulation of gene expression
0.5
regulation of protein metabolic process 0.6
multicellular organismal process regulation of cellular protein metabolic...
positive regulation of biosynthetic proc... 0.7
positive regulation of nucleobase-contai... cell proliferation cell surface receptor signaling pathway
transcription, DNA-templated negative regulation of metabolic process 0.8
cellular protein modification process positive regulation of cellular metaboli... 0.9
response to virus
innate immune response
positive regulation of RNA metabolic pro...
regulation of cellular macromolecule bio...
macromolecule modification 1.0
regulation of molecular function
cellular response to chemical stimulus
regulation of response to stimulus
regulation of cytokine-mediated signalin... RNA biosynthetic process positive regulation of nitrogen compound...
I-kappaB kinase/NF-kappaB signaling protein modification process positive regulation of macromolecule met... response to endogenous stimulus
positive regulation of macromolecule bio...
defense response to virus
regulation of cellular catabolic process positive regulation of cellular process
regulation of response to cytokine stimu... immune system process
regulation of catabolic process
process utilizing autophagic mechanism protein metabolic process cell death
aromatic compound biosynthetic process
regulation of innate immune response cellular response to stimulus cellular response to organic substance
nucleobase-containing compound biosynthe... regulation of cellular process positive regulation of biological proces...
regulation of cellular component organiz...
cellular nitrogen compound biosynthetic ... autophagy

cellular catabolic process biological regulation
positive regulation of RNA biosynthetic ... response to biotic stimulus localization
regulation of I-kappaB kinase/NF-kappaB ...
signal transduction negative regulation of cellular process regulation of response to stress
positive regulation of transcription by ... negative regulation of biological proces... regulation of cell differentiation
catabolic process regulation of biological process cellular component organization multi-organism process
regulation of DNA binding transcription ... organic cyclic compound biosynthetic pro... response to stimulus
positive regulation of nucleic acid-temp...
heterocycle biosynthetic process
activation of innate immune response positive regulation of developmental pro...
organelle organization monosaccharide metabolic process
positive regulation of DNA binding trans... positive regulation of response to stimu... regulation of cell communication
cell communication positive regulation of multicellular org...
positive regulation of transcription, DN... cellular macromolecule metabolic process cellular component organization or bioge... regulation of cell death
cardiac chamber development
neural tube development organonitrogen compound metabolic proces...
regulation of cysteine-type endopeptidas... heart development
positive regulation of cellular protein ... signaling
cell-cell signaling by wnt cell projection organization developmental process
regulation of multicellular organismal p...
cellular response to external stimulus embryo development positive regulation of cell communicatio...
regulation of developmental process movement of cell or subcellular componen...
cellular response to extracellular stimu... cardiac septum development cellular localization apoptotic signaling pathway
cell fate commitment
cellular response to organic cyclic comp... regulation of signaling
DNA-templated transcription, initiation cellular component assembly regulation of signal transduction
negative regulation of cell differentiat... heart morphogenesis positive regulation of signal transducti... cellular developmental process
positive regulation of signaling
maintenance of cell number
peptidyl-serine modification pattern specification process regulation of lipid metabolic process
carbohydrate homeostasis response to steroid hormone
embryonic morphogenesis phosphorus metabolic process
regulation of protein modification proce...
cellular process
regulation of cell cycle cellular component biogenesis
transmembrane receptor protein serine/th... cellular response to steroid hormone sti... regulation of small molecule metabolic p...
hormone-mediated signaling pathway small molecule biosynthetic process positive regulation of cellular protein ...
intracellular receptor signaling pathway membrane docking positive regulation of protein metabolic...
cilium assembly
cell cycle G2/M phase transition
protein phosphorylation
organelle localization negative regulation of developmental pro...
cilium organization
regulation of phosphate metabolic proces...
regulation of G2/M transitionregulation of mitotic cell cycle phase t...
of mitotic... negative regulation of cell communicatio... regulation of phosphorus metabolic proce...
regulation of primary metabolic process
regulation of microtubule-based process tube development
G2/M transition of mitotic cell cycle negative regulation of response to stimu...
regulation of cell cycle G2/M phase tran... cell projection assembly
organelle assembly
negative regulation of signaling regulation of macromolecule metabolic pr...
growth regulation of metabolic process
negative regulation of multicellular org...
microtubule cytoskeleton organization
plasma membrane bounded cell projection ...
regulation of cellular metabolic process
[3029, 4809.2) cytoskeleton organization
nucleotide catabolic process
microtubule-based process
ATP generation from ADP
cell cycle
glycolytic process
cell division nucleoside phosphate catabolic process nucleotide phosphorylation
cell cycle process

nucleoside monophosphate biosynthetic pr... ADP metabolic process
[1622.6, 2365.7) purine nucleoside monophosphate pyridine

biosynth...
nucleotide metabolic process
nucleoside triphosphate biosynthetic pro...

pyridine-containing compound metabolic p...
nucleoside diphosphate metabolic process
purine ribonucleoside triphosphate biosy...
pyruvate metabolic process
[702, 1304.5) ATP biosynthetic process

pyridine-containing compound biosyntheti...
pyridine nucleotide biosynthetic process
[215.2, 350.1)
[64, 131.8)
10
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
metabolic process
cellular metabolic process
primary metabolic process
negative regulation of macromolecule bio... organonitrogen compound metabolic proces...

protein modification process
organic substance catabolic process
negative regulation of biosynthetic proc...
organic substance metabolic process
negative regulation of cellular biosynth...
macromolecule modificationcellular protein modification process

cellular catabolic process
negative regulation of nucleic acid-temp...
macromolecule metabolic process
negative regulation of RNA biosynthetic
negative...regulation of cellular macromol...
nitrogen compound metabolic process DNA-templated transcription, initiation
hormone-mediated signaling pathway

negative regulation of RNA metabolic pro... catabolic process protein metabolic process
negative regulation of nucleobase-contai... intracellular receptor signaling pathway

cellular macromolecule metabolic process protein localization to organelle
proteolysis involved in cellular protein... interspecies interaction between organis...
cellular protein metabolic process
chromatin assembly or disassembly negative regulation of transcription, DN... proteolysis
nucleosome organization cellular response to steroid hormone sti...
cellular macromolecule localization
viral process
protein-DNA complex subunit organization protein polyubiquitination regulation of intracellular protein tran...
reproduction
modification-dependent protein catabolic... regulation of gene expression
organonitrogen compound catabolic proces...
chromosome organization regulation of catabolic process cellular response to organic cyclic comp...
negative regulation of transcription by ... cellular macromolecule catabolic process regulation of cellular biosynthetic proc...
protein-DNA complex assembly
regulation of gene expression, epigeneti... gene silencing regulation of intracellular transport
developmental process involved in reprod...
protein modification by small protein co...
process utilizing autophagic mechanism reproductive process
chromatin organization macromolecule catabolic process regulation of biosynthetic process
macromolecule methylation macromolecule biosynthetic process
peptidyl-lysine modification
regulation of protein catabolic process regulation of cellular protein localizat...
chromatin assembly protein modification by small protein re... multicellular organism reproduction
negative regulation of gene expression,
chromatin remodeling ...
negative regulation of metabolic process protein catabolic process
methylation myeloid cell differentiation aromatic compound biosynthetic process
negative regulation of gene expression regulation of macromolecule biosynthetic...
regulation of nucleobase-containing comp... gene silencing by miRNA
chromatin silencing megakaryocyte differentiation
regulation of histone modification regulation of gene silencing negative regulation of macromolecule met... modification-dependent macromolecule cat... catabolic process
proteasomal protein regulation of cellular protein catabolic... regulation of hemopoiesis
protein alkylation regulation of myeloid cell differentiati...
apoptotic signaling pathway regulation of gene silencing by miRNA nucleobase-containing compound
positive regulation of protein catabolic... nucleobase-containing compound
nucleicmetabolic...
acid metabolic process organic cyclic biosynthe...
compound biosynthetic pro...
cellular protein catabolic process
covalent chromatin modification
protein methylation protein acetylation intrinsic apoptotic signaling pathwayregulation of cellular response to stres...
macromolecule deacylation negative regulation of nitrogen compound...
cellular nitrogen compound biosynthetic ... Wnt signaling pathway
histone modification protein acylationprotein deacylation heterocycle biosynthetic process
positive regulation of biosynthetic proc... signal transduction by p53 class mediato...
positive regulation of macromolecule bio... heterocycle metabolic process
regulation of signal transduction by p53...
cell-cell signaling by wnt
response to UV cell surface receptor signaling pathway ... cellular aromatic compound metabolic pro...
positive regulation of gene expression regulation of Wnt signaling pathway
cell cycle response to ionizing radiation positive regulation of RNA biosynthetic ... regulation of cyclin-dependent protein k...
positive regulation of nucleic acid-temp... response to stress
cell cycle process
DNA repair positive regulation ofpositive
nucleobase-contai...
regulation of cellular biosynth...
positive regulation of RNA metabolic pro... positive regulation of macromolecule
nitrogenmet...
compound transport
cellular response to stress organic cyclic compound metabolic proces...
maintenance of cell number

gamete generation
response to abiotic stimulus regulation of cellular component organiz...
cellular process
cellular response to DNA damage stimulus peptide transport cellular protein-containing complex asse...
positive regulation of transcription, DN... immune system development
growth pattern specification process
negative regulation of cell cycle meiotic cell cycle process positive regulation of cellular metaboli...
germ cell development cellular response to oxygen levels
cell cycle phase transition positive regulation of nitrogen compound... rhythmic process
regulation of cell cycle organelle organization
embryo development
positive regulation of cell cycle reproductive structure development
meiotic cell cycle regulation of cellular macromolecule bio...
regulation of binding
reproductive system development macromolecule localization
cellular component organization regulation of protein stability
response to radiation
positive regulation of DNA metabolic pro...
anatomical structure homeostasis multicellular organism developmentmulti-organism process
regulation of RNA biosynthetic process
cellular
meiosis I cell cycle process involved in reproductio...
process positive regulation of metabolic process
positive regulation of response to DNA d...
RNA biosynthetic process
stem cell population maintenance
regulation of DNA metabolic process regulation of biological process cell aging
DNA biosynthetic process regulation of microtubule cytoskeleton o... cellular component organization or bioge...
regulation of microtubule-based process cell proliferation
signal transduction involved in cell cyc... cellular response to stimulus
organelle fission
cell cycle DNA replication
regulation of RNA metabolic process regulation of cyclin-dependent protein s...
signal transduction in response to DNA d... activation of innate immune response
transcription, DNA-templated biological regulation
regulation of DNA replication negative regulation of signal transducti...
microtubule cytoskeleton organization
regulation of nitrogen compound metaboli... regulation of transferase activityregulation of cellular process
negative regulation of DNA metabolic
DNA pro...
[3076.4, 4659.7) metabolic process negative regulation of chromosome segreg...
DNA replication
cell division chromosome separation
protein localization to chromosome
chromosome localization
regulation of nuclear division regulation of cellular metabolic process negative regulation of cellular process
establishment of chromosome localization

establishment of organelle localization regulation of mitotic nuclear division
[1090.8, 1676.7) negative regulation of organelle organiz... regulation of primary metabolic process
multivesicular body assembly positive regulation of biological proces...

negative regulation of cell cycle proces...
chromosome segregation cell cycle checkpoint negative regulation of biological proces...
nuclear chromosome segregation
viral budding mitotic sister chromatid segregation

multivesicular body organization
positive regulation of cellular process
[394.4, 647.5)
virion assembly regulation of chromosome segregation
regulation of sister chromatid segregati...
[178.2, 244.9)
[23, 96.1)
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
organophosphate biosynthetic process
nucleoside phosphate metabolic process
carbohydrate derivative biosynthetic pro...

heterocycle metabolic process
carbohydrate derivative metabolic proces...
ribose phosphate metabolic process

ribose phosphate biosynthetic process
organophosphate metabolic process
purine-containing compound metabolic pro...

purine-containing compound biosynthetic ... cellular metabolic process biosynthetic process
[4717.6, 5041)
nucleoside phosphate biosynthetic proces...
cellular nitrogen compound metabolic pro...metabolic process

nucleobase-containing compound metabolic... purine-containing compound salvage
nucleotide salvage
[1126.4, 3401.8) heterocycle biosynthetic process
cellular biosynthetic process
glycosyl compound metabolic process
pyrimidine-containing compound biosynthe... small molecule metabolic process

nucleobase-containing compound biosynthe... pyrimidine-containing compound catabolic... nucleobase-containing small molecule bio...
[272.8, 569) organic substance biosynthetic process glycosyl compound biosynthetic process
nucleobase-containing small molecule cat...
organonitrogen compound biosynthetic pro... nucleoside catabolic process
cellular metabolic compound salvage
pyrimidine-containing compound metabolic...
nucleoside biosynthetic process
nucleoside metabolic process
snRNA metabolic process
[42.8, 121.6) aromatic compound biosynthetic process
nucleobase metabolic process
[13, 34.4)
Frontiers 11
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
mRNA processing
protein import into nucleus
protein import
nuclear transport
establishment of RNA localization
[4392.4, 4801.8) RNA processing
establishment of protein localization to...
mRNA export from nucleus
intracellular transport
peptide transport
nucleobase-containing compound metabolic... protein-containing complex localization

gene expression
[1599, 2261.2)
mRNA metabolic process
RNA splicing
ribonucleoprotein complex localization

[414.6, 692)
nucleic acid metabolic process nucleobase-containing compound transport

RNA localization
heterocycle metabolic process

[199.2, 255.8)
[102, 154.2)
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
biosynthetic process aromatic compound catabolic process
RNA catabolic cellular nitrogen compound catabolic pro...
process of mRNA metabolic process
regulation
cellular biosynthetic process
organic cyclic compound metabolic proces... regulation of RNA stability organic cyclic compound catabolic proces...
nucleobase-containing compound catabolic...
cellular catabolic process mRNA catabolic process protein localization to endoplasmic reti...
gene silencing protein targeting to ER
organic substance catabolic process cellular nitrogen compound metabolic pro... heterocycle catabolic process
negative regulation of metabolic process
RNA localization
negative regulation of gene expression nucleobase-containing compound transport cytoplasmic translation
RNA phosphodiester bond hydrolysis regulation of mRNA catabolic process
organelle assembly
negative regulation of macromolecule met... cotranslational protein targeting to mem...
cellular macromolecule catabolic process gene expression
ribonucleoprotein complex export from nu... mRNA metabolic process
protein targeting organic substance biosynthetic process heterocycle metabolic process establishment of protein localization to...
protein-containing complex localization nuclear-transcribed mRNA catabolic proce...
nucleic acid phosphodiester bond hydroly...
ribonucleoprotein complex localization macromolecule biosynthetic process protein targeting to membrane

organic substance transport
establishment of protein localization to... establishment of RNA localization translational initiation
protein localization to membrane nucleic acid metabolic process
macromolecule catabolic process cellular localization
establishment of protein localization to...
catabolic process
protein localization to organelle
ncRNA processing amide biosynthetic process
organonitrogen compoundnucleobase-containing
biosynthetic pro... compound metabolic...
intracellular protein transport
protein-containing complex subunit organ... peptide transport
rRNA processing ribonucleoprotein complex biogenesis
cellular component biogenesis nitrogen compound transport
cellular amide metabolic process cellular macromolecule biosynthetic proc... establishment of protein localization
ncRNA metabolic process
ribosome biogenesis peptide metabolic process
cellular protein-containing complex asse...
[3932, 5029) rRNA metabolic process cellular macromolecule localization intracellular transport
ribonucleoprotein complex subunit organi...
macromolecule localization
RNA metabolic process
RNA processing ribonucleoprotein complex assembly posttranscriptional regulation of gene e... negative regulation of protein-containing complex assembly
biological proces...
peptide biosynthetic process cellular protein localization

cellular component organization or bioge...
regulation of translation
[1533, 1939.5) regulation of cellular amide metabolic p... macromolecule metabolic process

translation
primary metabolic process
[526, 772)
nitrogen compound metabolic process
cellular metabolic process
[213, 326.5) metabolic process
[88, 147)
12
0.0

0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
nucleoside phosphate metabolic process 1.0
purine nucleotide metabolic process

nucleoside monophosphate metabolic proce...
[927.4, 1022)
ATP metabolic process
oxidation-reduction process ribose phosphate metabolic process
small molecule metabolic process

cellular respiration
oxidative phosphorylation
[552.6, 744.7) carbohydrate derivative metabolic proces...
generation of precursor metabolites and ...
electron transport chain
purine-containing compound metabolic pro...
transmembrane transport
nucleoside triphosphate metabolic proces...

ion transmembrane transport
[407.6, 422.5) energy derivation by oxidation of organi...

organophosphate metabolic process
drug metabolic process
[229.6, 279.3)
[113, 164.7)
Frontiers 13
Adjacency Matrix of Interactions Between Clusters

Time-Interval 1
Cluster
1 2 3 4 5 6 7 8 9 10 11
344 2 0 0 0 21 40 2 2 0 0
1
2 107 8 3 0 6 11 0 0 0 0
2
0 8 130 5 38 78 36 4 1 6 0
3
0 3 5 393 38 128 101 16 15 21 8

4
0 0 38 38 1244 1101 686 56 35 26 18

5
Cluster
21 6 78 128 1101 12479 10485 490 449 794 129

6
40 11 36 101 686 10485 79128 2674 2269 1616 86

7
2 0 4 16 56 490 2674 2217 905 484 59

8
2 0 1 15 35 449 2269 905 1524 692 58

9
11 10
0 0 6 21 26 794 1616 484 692 17467 94
0 0 0 8 18 129 86 59 58 94 707
Time-Interval 2
Cluster
1 2 3 4 5 6 7 8 9 10 11
12630 4636 828 1053 102 844 54 209 127 66 2
1
4636 59802 7791 5184 470 2418 95 559 186 81 9

2
828 7791 7090 5299 342 1913 52 180 61 28 4

3
1053 5184 5299 14532 1588 5060 177 444 222 149 7
4
102 470 342 1588 925 3712 49 480 83 30 3

5
Cluster
844 2418 1913 5060 3712 91802 2377 8090 454 642 56
6
54 95 52 177 49 2377 767 984 141 65 1

7
209 559 180 444 480 8090 984 43211 1492 291 26
8
127 186 61 222 83 454 141 1492 2685 1488 10

9
11 10
66 81 28 149 30 642 65 291 1488 1006 41
2 9 4 7 3 56 1 26 10 41 182
Figure S18: Adjacency matrix used to draw the internal and external connectivity lines of PPI networks of
time-interval 1 and 2, shown in Figure 1b) and 1d), respectively
14
SUPPLEMENTARY TABLES S1 AND S2

Tables S1 and S2 (inside file Table1.XLSX) contain the list of enriched terms of time-interval 1 and 2
clusters, respectively, as described below:
Field Description
GO.ID Gene Ontology term ID
Term Human-readable term name
Annotated Number of genes annotated for a specific GO term
Significant Number of differentially expressed genes identified
Expected Number of differentially expressed genes expected by the topGO statistical test
pValue p-values adjusted by the Benjamini-Hochberg procedure.
Frontiers 15
List of Enriched Terms - Time-Interval 1
GO:0031326 regulation of cellular biosynthetic proc... 3338 235 73.78 4.98354838709678e-28
GO:0009889 regulation of biosynthetic process 3399 235 75.13 4.98354838709678e-28
GO:0031323 regulation of cellular metabolic process 4883 238 107.93 4.98354838709678e-28
GO:0019219 regulation of nucleobase-containing comp... 3217 236 71.11 4.98354838709678e-28
GO:0051252 regulation of RNA metabolic process 2978 236 65.82 4.98354838709678e-28
GO:0009059 macromolecule biosynthetic process 4059 235 89.72 4.98354838709678e-28
GO:0010556 regulation of macromolecule biosynthetic... 3207 235 70.89 4.98354838709678e-28
GO:0090304 nucleic acid metabolic process 4138 239 91.46 4.98354838709678e-28
GO:0006351 transcription, DNA-templated 2873 235 63.5 4.98354838709678e-28
GO:0006139 nucleobase-containing compound metabolic... 4630 241 102.34 4.98354838709678e-28
GO:0018130 heterocycle biosynthetic process 3399 235 75.13 4.98354838709678e-28
GO:0034645 cellular macromolecule biosynthetic proc... 3932 235 86.91 4.98354838709678e-28
GO:0019222 regulation of metabolic process 5251 241 116.07 4.98354838709678e-28
GO:0032774 RNA biosynthetic process 2917 235 64.48 4.98354838709678e-28
GO:0019438 aromatic compound biosynthetic process 3406 235 75.28 4.98354838709678e-28
GO:0006725 cellular aromatic compound metabolic pro... 4819 242 106.52 4.98354838709678e-28
GO:0046483 heterocycle metabolic process 4776 241 105.57 4.98354838709678e-28
GO:1901362 organic cyclic compound biosynthetic pro... 3535 235 78.14 4.98354838709678e-28
GO:1901360 organic cyclic compound metabolic proces... 5009 242 110.72 4.98354838709678e-28
GO:0010468 regulation of gene expression 3522 238 77.85 4.98354838709678e-28
GO:0080090 regulation of primary metabolic process 4805 239 106.21 4.98354838709678e-28
GO:0051171 regulation of nitrogen compound metaboli... 4663 239 103.07 4.98354838709678e-28
1 GO:0010467 gene expression 4333 238 95.77 4.98354838709678e-28
GO:0034641 cellular nitrogen compound metabolic pro... 5227 241 115.54 4.98354838709678e-28
GO:0044249 cellular biosynthetic process 4972 236 109.9 4.98354838709678e-28
GO:1901576 organic substance biosynthetic process 5049 236 111.6 4.98354838709678e-28
GO:0009058 biosynthetic process 5106 236 112.86 4.98354838709678e-28
GO:0034654 nucleobase-containing compound biosynthe... 3345 235 73.94 4.98354838709678e-28
GO:0060255 regulation of macromolecule metabolic pr... 4858 240 107.38 4.98354838709678e-28
GO:0044271 cellular nitrogen compound biosynthetic ... 3956 235 87.44 4.98354838709678e-28
GO:2000112 regulation of cellular macromolecule bio... 3101 235 68.54 4.98354838709678e-28
GO:0006355 regulation of transcription, DNA-templat... 2729 235 60.32 6.7589375e-28
GO:2001141 regulation of RNA biosynthetic process 2778 235 61.4 4.58788484848485e-24
GO:0044260 cellular macromolecule metabolic process 6674 248 147.52 5.45258823529412e-22
GO:0050789 regulation of biological process 9094 262 201.01 4.8554e-14
GO:0050794 regulation of cellular process 8535 252 188.65 1.02993333333333e-13
GO:0044237 cellular metabolic process 8695 253 192.19 3.21506216216216e-12
GO:0043170 macromolecule metabolic process 7510 250 166 1.38227894736842e-11
GO:0008152 metabolic process 9233 259 204.08 2.81251025641026e-11
GO:0006357 regulation of transcription by RNA polym... 2066 216 45.67 3.2829125e-11
GO:0065007 biological regulation 9603 265 212.26 3.31588292682927e-11
GO:0016070 RNA metabolic process 3693 238 81.63 7.7245e-09
GO:0006366 transcription by RNA polymerase II 2189 216 48.38 2.31735e-08
GO:0006807 nitrogen compound metabolic process 8145 253 180.03 5.49297777777778e-06
3 GO:0051259 protein complex oligomerization 445 17 6.07 2.93531e-05
GO:0055085 transmembrane transport 1229 42 16.77 0.0003321535
GO:0043062 extracellular structure organization 351 30 4.06 5.329905e-13
GO:0022610 biological adhesion 1120 49 12.96 5.329905e-13
GO:1901615 organic hydroxy compound metabolic proce... 445 21 5.15 5.92211666666667e-06
GO:0006066 alcohol metabolic process 296 19 3.43 5.92211666666667e-06
GO:0051056 regulation of small GTPase mediated sign... 269 20 3.11 2.5104625e-05
4 GO:0050817 coagulation 308 18 3.56 0.000126400909090909
GO:0007264 small GTPase mediated signal transductio... 466 23 5.39 0.00015449
GO:0009611 response to wounding 562 23 6.5 0.000166373846153846
GO:0023052 signaling 5319 93 61.55 0.000275875
GO:0007154 cell communication 5349 94 61.89 0.0003476025
GO:0009653 anatomical structure morphogenesis 2056 44 23.79 0.000699270526315789
GO:0016050 vesicle organization 278 48 5.16 6.64307e-22
GO:0006903 vesicle targeting 85 23 1.58 1.23592e-16
GO:0140029 exocytic process 66 20 1.22 1.59639666666667e-15
GO:0022406 membrane docking 159 27 2.95 7.7245e-15
GO:0048278 vesicle docking 58 26 1.08 8.34246e-15
GO:0048284 organelle fusion 94 24 1.74 3.0898e-13
5 GO:0051179 localization 5214 159 96.68 1.78767e-12
GO:0048208 COPII vesicle coating 63 20 1.17 2.5104625e-12
GO:0061024 membrane organization 668 53 12.39 4.11973333333333e-12
GO:0016192 vesicle-mediated transport 1644 102 30.49 5.25266e-12
GO:0006906 vesicle fusion 72 24 1.34 7.16271818181818e-10
GO:0051650 establishment of vesicle localization 221 35 4.1 3.80283076923077e-08
GO:0006888 ER to Golgi vesicle-mediated transport 188 33 3.49 4.8554e-08
GO:0051640 organelle localization 565 53 10.48 1.25523125e-06
GO:0051656 establishment of organelle localization 398 37 7.38 2.81717058823529e-06
GO:0006900 vesicle budding from membrane 93 21 1.72 5.23549444444444e-06
GO:0046907 intracellular transport 1531 83 28.39 1.30096842105263e-05
GO:0032940 secretion by cell 1227 52 22.75 1.467655e-05
GO:0048193 Golgi vesicle transport 320 43 5.93 3.16336666666667e-05
GO:0061025 membrane fusion 148 29 2.74 4.42403181818182e-05
GO:0010941 regulation of cell death 1376 57 25.52 5.27840833333333e-05
GO:0051641 cellular localization 2367 105 43.89 5.27840833333333e-05
GO:0051649 establishment of localization in cell 1838 91 34.08 9.2694e-05
GO:0051173 positive regulation of nitrogen compound... 2648 77 49.1 9.50707692307692e-05
5 GO:0002376 immune system process 2380 77 44.13 0.000125880740740741
GO:0023052 signaling 5319 138 98.63 0.0001489725
GO:0007154 cell communication 5349 139 99.19 0.000298680666666667
GO:0010604 positive regulation of macromolecule met... 2751 77 51.01 0.000598025806451613
GO:0008219 cell death 1804 62 33.45 0.00067589375
GO:0019438 aromatic compound biosynthetic process 3406 67 63.16 0.000749042424242424
GO:0010942 positive regulation of cell death 562 31 10.42 0.00075038
GO:0018130 heterocycle biosynthetic process 3399 67 63.03 0.00075038
GO:1901362 organic cyclic compound biosynthetic pro... 3535 69 65.55 0.00077245
GO:0010952 positive regulation of peptidase activit... 157 16 2.91 0.000835081081081081
GO:0001775 cell activation 1152 45 21.36 0.000853760526315789
GO:0060548 negative regulation of cell death 828 38 15.35 0.000990320512820513
GO:0044782 cilium organization 297 140 18.68 1.40445454545455e-27
GO:0007017 microtubule-based process 602 170 37.87 1.40445454545455e-27
GO:0022406 membrane docking 159 78 10 1.40445454545455e-27
GO:0070925 organelle assembly 670 175 42.14 1.40445454545455e-27
GO:0030031 cell projection assembly 448 143 28.18 1.40445454545455e-27
GO:0009893 positive regulation of metabolic process 2921 340 183.73 1.40445454545455e-27
GO:0031325 positive regulation of cellular metaboli... 2736 329 172.09 1.40445454545455e-27
GO:0010604 positive regulation of macromolecule met... 2751 325 173.03 1.40445454545455e-27
GO:0060271 cilium assembly 289 138 18.18 1.40445454545455e-27
GO:0048518 positive regulation of biological proces... 4802 465 302.04 1.40445454545455e-27
GO:0007049 cell cycle 1522 216 95.73 8.31869230769231e-27
GO:0048522 positive regulation of cellular process 4294 438 270.09 8.31869230769231e-27
GO:0023052 signaling 5319 487 334.56 7.51851333333333e-25
GO:0097711 ciliary basal body-plasma membrane docki... 86 76 5.41 1.7380125e-24
GO:0120031 plasma membrane bounded cell projection ... 439 143 27.61 5.72521764705882e-24
GO:0007165 signal transduction 4969 477 312.54 8.23946666666667e-24
GO:0140056 organelle localization by membrane tethe... 150 78 9.43 4.22814736842105e-23
GO:0030030 cell projection organization 1197 203 75.29 6.1796e-23
GO:0022402 cell cycle process 1118 167 70.32 2.80890909090909e-22
GO:0006367 transcription initiation from RNA polyme... 165 59 10.38 1.74640869565217e-21
6 GO:0007154 cell communication 5349 495 336.45 2.5104625e-21
GO:0008219 cell death 1804 225 113.47 2.16286e-20
GO:0010033 response to organic substance 2704 312 170.08 1.78257692307692e-19
GO:0050789 regulation of biological process 9094 693 572 3.26145555555556e-19
GO:0051726 regulation of cell cycle 983 161 61.83 2.45053103448276e-18
GO:0048519 negative regulation of biological proces... 4189 388 263.48 7.20953333333334e-18
GO:0071840 cellular component organization or bioge... 5277 459 331.92 2.655296875e-17
GO:0030705 cytoskeleton-dependent intracellular tra... 150 49 9.43 7.02227272727273e-17
GO:0060255 regulation of macromolecule metabolic pr... 4858 429 305.56 1.95384411764706e-16
GO:0007389 pattern specification process 349 77 21.95 3.48706e-16
GO:0031328 positive regulation of cellular biosynth... 1655 208 104.1 5.14966666666667e-16
GO:0099111 microtubule-based transport 134 44 8.43 5.42802702702703e-16
GO:0007018 microtubule-based movement 229 76 14.4 8.71482051282051e-16
GO:0009891 positive regulation of biosynthetic proc... 1683 208 105.86 1.4290325e-15
GO:0006352 DNA-templated transcription, initiation 211 59 13.27 8.26341860465116e-15
GO:0010557 positive regulation of macromolecule bio... 1575 201 99.07 8.26341860465116e-15
GO:0016043 cellular component organization 5129 456 322.61 1.18332765957447e-14
GO:0051716 cellular response to stimulus 6086 517 382.8 1.38397291666667e-14
GO:0050896 response to stimulus 7323 575 460.61 3.0898e-14
GO:0045935 positive regulation of nucleobase-contai... 1588 197 99.88 5.14966666666667e-14
GO:0070887 cellular response to chemical stimulus 2671 305 168 1.36664230769231e-12
GO:0010389 regulation of G2/M transition of mitotic... 139 61 8.74 1.51575094339623e-12
GO:0051179 localization 5214 436 327.95 1.51629074074074e-12
GO:0010941 regulation of cell death 1376 182 86.55 2.58419636363636e-12
GO:0032502 developmental process 4760 405 299.4 2.593225e-12
GO:0000086 G2/M transition of mitotic cell cycle 182 67 11.45 3.25242105263158e-12
GO:0048523 negative regulation of cellular process 3779 351 237.69 3.72906896551724e-12
GO:0048584 positive regulation of response to stimu... 1878 223 118.12 4.1895593220339e-12
GO:0051640 organelle localization 565 117 35.54 5.66463333333333e-12
GO:0051246 regulation of protein metabolic process 2330 252 146.55 8.61091803278689e-12
GO:0030522 intracellular receptor signaling pathway 257 65 16.16 9.71791935483871e-12
GO:0044839 cell cycle G2/M phase transition 199 68 12.52 1.59394444444444e-11
GO:1903508 positive regulation of nucleic acid-temp... 1357 177 85.35 3.37946875e-11
GO:0006793 phosphorus metabolic process 2768 260 174.1 6.78819696969697e-11
GO:0009755 hormone-mediated signaling pathway 217 57 13.65 7.37862686567164e-11
GO:0048583 regulation of response to stimulus 3365 333 211.65 7.49730882352941e-11
GO:0061919 process utilizing autophagic mechanism 392 67 24.66 9.85153623188406e-11
GO:0051254 positive regulation of RNA metabolic pro... 1428 180 89.82 1.28006e-10
GO:0045944 positive regulation of transcription by ... 989 137 62.21 1.45786338028169e-10
GO:0022607 cellular component assembly 2476 288 155.74 1.63072777777778e-10
GO:1902749 regulation of cell cycle G2/M phase tran... 154 61 9.69 2.32793150684932e-10
GO:1902680 positive regulation of RNA biosynthetic ... 1358 177 85.42 3.21018181818182e-10
GO:0051607 defense response to virus 201 47 12.64 3.21018181818182e-10
GO:0071383 cellular response to steroid hormone sti... 227 62 14.28 3.91113924050633e-10
GO:0023051 regulation of signaling 2907 305 182.85 3.91113924050633e-10
GO:0009987 cellular process 12542 832 788.88 5.9864875e-10
GO:0048869 cellular developmental process 3276 303 206.06 6.29403703703704e-10
GO:0045595 regulation of cell differentiation 1377 172 86.61 6.40568292682927e-10
6 GO:0045893 positive regulation of transcription, DN... 1279 163 80.45 7.45187058823529e-10
GO:0032268 regulation of cellular protein metabolic... 2170 238 136.49 7.45187058823529e-10
GO:0009719 response to endogenous stimulus 1405 176 88.37 8.6226976744186e-10
GO:0042073 intraciliary transport 50 36 3.14 1.15423563218391e-09
GO:0051247 positive regulation of protein metabolic... 1428 178 89.82 1.931125e-09
GO:0031929 TOR signaling 94 33 5.91 2.23152222222222e-09
GO:0051239 regulation of multicellular organismal p... 2354 248 148.06 2.23152222222222e-09
GO:0000226 microtubule cytoskeleton organization 436 115 27.42 2.37676923076923e-09
GO:0023056 positive regulation of signaling 1470 188 92.46 2.49177419354839e-09
GO:0007166 cell surface receptor signaling pathway 2446 304 153.85 2.49177419354839e-09
GO:0031399 regulation of protein modification proce... 1515 190 95.29 3.45137234042553e-09
GO:0010646 regulation of cell communication 2879 304 181.09 3.57766315789474e-09
GO:0050793 regulation of developmental process 1985 218 124.85 3.70132291666667e-09
GO:0071310 cellular response to organic substance 2238 285 140.77 4.30023711340206e-09
GO:0007507 heart development 432 77 27.17 4.72928571428571e-09
GO:0007224 smoothened signaling pathway 106 39 6.67 5.61781818181818e-09
GO:0031023 microtubule organizing center organizati... 123 36 7.74 9.42389e-09
GO:0032886 regulation of microtubule-based process 177 43 11.13 1.04013069306931e-08
GO:0046031 ADP metabolic process 74 21 4.65 1.25712450980392e-08
GO:0010647 positive regulation of cell communicatio... 1464 188 92.08 1.45490582524272e-08
GO:0006096 glycolytic process 64 21 4.03 1.63402884615385e-08
GO:0060759 regulation of response to cytokine stimu... 139 40 8.74 2.33192452830189e-08
GO:0006468 protein phosphorylation 1629 194 102.46 2.33192452830189e-08
GO:0034340 response to type I interferon 83 31 5.22 2.88766355140187e-08
GO:0051174 regulation of phosphorus metabolic proce... 1456 180 91.58 3.00397222222222e-08
GO:0010628 positive regulation of gene expression 1621 182 101.96 3.25988073394495e-08
GO:0019362 pyridine nucleotide metabolic process 129 26 8.11 4.73212612612613e-08
GO:0043412 macromolecule modification 3579 309 225.11 4.73212612612613e-08
GO:0065009 regulation of molecular function 2633 266 165.61 5.1952389380531e-08
GO:0006928 movement of cell or subcellular componen... 1669 173 104.98 5.1952389380531e-08
GO:0009967 positive regulation of signal transducti... 1337 184 84.1 1.0434850877193e-07
GO:0009132 nucleoside diphosphate metabolic process 101 23 6.35 1.47773043478261e-07
GO:0035050 embryonic heart tube development 61 24 3.84 1.5981724137931e-07
GO:0032006 regulation of TOR signaling 78 27 4.91 3.16902564102564e-07
GO:0051240 positive regulation of multicellular org... 1315 158 82.71 3.66586440677966e-07
GO:0036211 protein modification process 3414 305 214.74 3.99099166666667e-07
GO:0006914 autophagy 392 67 24.66 3.99099166666667e-07
GO:0007249 I-kappaB kinase/NF-kappaB signaling 214 49 13.46 4.8517520661157e-07
GO:0071357 cellular response to type I interferon 80 31 5.03 5.40086991869919e-07
GO:0019216 regulation of lipid metabolic process 333 59 20.95 5.40086991869919e-07
GO:0044085 cellular component biogenesis 2670 291 167.94 6.47861290322581e-07
GO:0019827 stem cell population maintenance 125 30 7.86 1.0134544e-06
GO:0018209 peptidyl-serine modification 266 45 16.73 1.07897777777778e-06
GO:0006757 ATP generation from ADP 65 21 4.09 1.16779842519685e-06
GO:0002376 immune system process 2380 213 149.7 1.31735658914729e-06
GO:0048585 negative regulation of response to stimu... 1294 146 81.39 1.66373846153846e-06
GO:0032270 positive regulation of cellular protein ... 1338 166 84.16 1.88690076335878e-06
GO:0006754 ATP biosynthetic process 101 21 6.35 2.2237196969697e-06
GO:0032501 multicellular organismal process 5860 446 368.59 2.43931578947368e-06
GO:0007178 transmembrane receptor protein serine/th... 282 49 17.74 2.88227611940299e-06
GO:0023057 negative regulation of signaling 1106 136 69.57 3.43311111111111e-06
GO:0002218 activation of innate immune response 206 44 12.96 3.52146323529412e-06
GO:0009966 regulation of signal transduction 2612 294 164.29 4.05959124087591e-06
GO:0031503 protein-containing complex localization 239 46 15.03 4.1421231884058e-06
GO:0003007 heart morphogenesis 210 48 13.21 4.33461151079137e-06
GO:0060337 type I interferon signaling pathway 80 31 5.03 4.52435e-06
GO:0019363 pyridine nucleotide biosynthetic process 93 22 5.85 4.60182978723404e-06
GO:0097190 apoptotic signaling pathway 496 82 31.2 5.29371328671329e-06
GO:0032606 type I interferon production 99 30 6.23 5.71401369863014e-06
GO:0019538 protein metabolic process 4819 370 303.11 5.99042857142857e-06
GO:0001959 regulation of cytokine-mediated signalin... 129 37 8.11 7.30695945945946e-06
GO:0048545 response to steroid hormone 334 66 21.01 1.01610872483221e-05
GO:0019220 regulation of phosphate metabolic proces... 1448 179 91.08 1.22773509933775e-05
GO:1901292 nucleoside phosphate catabolic process 122 22 7.67 1.22773509933775e-05
GO:0006996 organelle organization 3158 331 198.63 1.52457236842105e-05
GO:0098727 maintenance of cell number 128 30 8.05 1.71655555555556e-05
GO:0051241 negative regulation of multicellular org... 919 110 57.8 1.79407741935484e-05
GO:0044380 protein localization to cytoskeleton 44 18 2.77 1.79407741935484e-05
6 GO:0009166 nucleotide catabolic process 116 22 7.3 1.88160897435897e-05
GO:0010648 negative regulation of cell communicatio... 1102 135 69.31 2.13759748427673e-05
GO:0051704 multi-organism process 1984 178 124.79 2.22079375e-05
GO:0051090 regulation of DNA binding transcription ... 360 61 22.64 2.27469938650307e-05
GO:0009142 nucleoside triphosphate biosynthetic pro... 128 22 8.05 2.27469938650307e-05
GO:0034219 carbohydrate transmembrane transport 99 16 6.23 2.27469938650307e-05
GO:0006090 pyruvate metabolic process 100 26 6.29 2.35503048780488e-05
GO:1901990 regulation of mitotic cell cycle phase t... 322 75 20.25 2.71527878787879e-05
GO:0046785 microtubule polymerization 59 20 3.71 3.88537724550898e-05
GO:0072524 pyridine-containing compound metabolic p... 134 26 8.43 4.23008333333333e-05
GO:0046902 regulation of mitochondrial membrane per... 59 16 3.71 4.38788165680473e-05
GO:0032479 regulation of type I interferon producti... 98 29 6.16 4.45294705882353e-05
GO:0033500 carbohydrate homeostasis 185 30 11.64 4.7882865497076e-05
GO:0003170 heart valve development 46 16 2.89 4.94008720930233e-05
GO:0071496 cellular response to external stimulus 274 51 17.23 5.9831387283237e-05
GO:0021915 neural tube development 127 31 7.99 6.03754022988506e-05
GO:0033209 tumor necrosis factor-mediated signaling... 102 34 6.42 6.1796e-05
GO:0010921 regulation of phosphatase activity 135 31 8.49 6.7589375e-05
GO:0018210 peptidyl-threonine modification 109 24 6.86 8.55368361581921e-05
GO:0008283 cell proliferation 1662 151 104.54 9.49379888268157e-05
GO:0044248 cellular catabolic process 1935 173 121.71 0.000102424309392265
GO:0009615 response to virus 275 54 17.3 0.000102424309392265
GO:0072525 pyridine-containing compound biosyntheti... 96 22 6.04 0.00011035
GO:0030856 regulation of epithelial cell differenti... 110 29 6.92 0.000118189071038251
GO:0051091 positive regulation of DNA binding trans... 228 44 14.34 0.000133612972972973
GO:2000116 regulation of cysteine-type endopeptidas... 200 38 12.58 0.000140445454545455
GO:0198738 cell-cell signaling by wnt 400 60 25.16 0.000155307407407407
GO:0046890 regulation of lipid biosynthetic process 156 31 9.81 0.000178883157894737
GO:0030509 BMP signaling pathway 123 28 7.74 0.000186035078534031
GO:0007010 cytoskeleton organization 1076 151 67.68 0.000224130569948187
GO:0031324 negative regulation of cellular metaboli... 2065 183 129.89 0.000237676923076923
GO:0060411 cardiac septum morphogenesis 64 19 4.03 0.000237676923076923
GO:0051301 cell division 538 64 33.84 0.000249680808080808
GO:0032480 negative regulation of type I interferon... 39 17 2.45 0.000249680808080808
GO:0043122 regulation of I-kappaB kinase/NF-kappaB ... 184 39 11.57 0.000256189447236181
GO:0045088 regulation of innate immune response 304 55 19.12 0.000262633
GO:0051093 negative regulation of developmental pro... 733 95 46.1 0.000269012437810945
GO:0071407 cellular response to organic cyclic comp... 502 92 31.58 0.000275328712871287
GO:0009206 purine ribonucleoside triphosphate biosy... 110 21 6.92 0.00028158275862069
GO:0040007 growth 764 80 48.05 0.000287775490196078
GO:0051094 positive regulation of developmental pro... 1072 132 67.43 0.000292481067961165
GO:0005996 monosaccharide metabolic process 255 33 16.04 0.000326805769230769
GO:0080134 regulation of response to stress 1136 137 71.45 0.000326805769230769
GO:0062012 regulation of small molecule metabolic p... 322 49 20.25 0.000362201435406699
GO:0045087 innate immune response 710 89 44.66 0.000382546666666667
GO:1903829 positive regulation of cellular protein ... 281 49 17.67 0.00040808679245283
GO:0045445 myoblast differentiation 73 20 4.59 0.00040808679245283
GO:0006464 cellular protein modification process 3414 305 214.74 0.000413423943661972
GO:0007219 Notch signaling pathway 156 39 9.81 0.00044758785046729
GO:0002065 columnar/cuboidal epithelial cell differ... 92 25 5.79 0.000452691627906977
GO:0051128 regulation of cellular component organiz... 2018 202 126.93 0.000457748148148148
GO:0090559 regulation of membrane permeability 65 16 4.09 0.00047480871559633
GO:1901564 organonitrogen compound metabolic proces... 5705 412 358.84 0.000498581363636364
GO:0044283 small molecule biosynthetic process 659 72 41.45 0.000521925675675676
GO:0007098 centrosome cycle 112 34 7.04 0.000533440807174888
GO:0009894 regulation of catabolic process 732 93 46.04 0.00053795625
GO:0031329 regulation of cellular catabolic process 644 85 40.51 0.000567374778761062
6 GO:0003279 cardiac septum development 87 21 5.47 0.000571681057268722
GO:1903901 negative regulation of viral life cycle 67 16 4.21 0.000586927074235808
GO:0046939 nucleotide phosphorylation 82 22 5.16 0.000597809130434783
GO:0045596 negative regulation of cell differentiat... 537 75 33.78 0.000601909090909091
GO:0009056 catabolic process 2174 182 136.74 0.000665905172413793
GO:0009790 embryo development 772 98 48.56 0.000714029411764706
GO:0045165 cell fate commitment 203 38 12.77 0.000714029411764706
GO:0042594 response to starvation 154 31 9.69 0.000714029411764706
GO:0035295 tube development 833 104 52.39 0.000714029411764706
GO:0009892 negative regulation of metabolic process 2327 195 146.37 0.000714029411764706
GO:0006355 regulation of transcription, DNA-templat... 2729 251 171.65 0.000775682008368201
GO:0009127 purine nucleoside monophosphate biosynth... 126 21 7.93 0.000836820833333333
GO:0006820 anion transport 448 32 28.18 0.000897452282157676
GO:0003205 cardiac chamber development 133 27 8.37 0.00093066265060241
GO:0009607 response to biotic stimulus 790 94 49.69 0.00093066265060241
GO:0048598 embryonic morphogenesis 465 68 29.25 0.00093066265060241
GO:0035735 intraciliary transport involved in ciliu... 39 31 2.45 0.00093066265060241
GO:0031668 cellular response to extracellular stimu... 217 37 13.65 0.00093066265060241
GO:0008593 regulation of Notch signaling pathway 75 22 4.72 0.00093066265060241
GO:0030178 negative regulation of Wnt signaling pat... 139 34 8.74 0.00093066265060241
GO:0009124 nucleoside monophosphate biosynthetic pr... 148 21 9.31 0.000984796812749004
GO:0051276 chromosome organization 1027 621 130.41 2.71035087719298e-28
GO:0007049 cell cycle 1522 579 193.27 2.71035087719298e-28
GO:0006325 chromatin organization 667 401 84.7 2.71035087719298e-28
GO:0070647 protein modification by small protein co... 904 544 114.79 2.71035087719298e-28
GO:0022402 cell cycle process 1118 464 141.97 2.71035087719298e-28
GO:0033554 cellular response to stress 1625 552 206.35 2.71035087719298e-28
GO:0006259 DNA metabolic process 858 472 108.95 2.71035087719298e-28
GO:0071824 protein-DNA complex subunit organization 226 171 28.7 2.71035087719298e-28
GO:0044257 cellular protein catabolic process 664 353 84.32 2.71035087719298e-28
GO:0030163 protein catabolic process 792 373 100.57 2.71035087719298e-28
GO:0007059 chromosome segregation 269 167 34.16 2.71035087719298e-28
7 GO:0051603 proteolysis involved in cellular protein... 618 350 78.48 2.71035087719298e-28
GO:0051726 regulation of cell cycle 983 370 124.83 2.71035087719298e-28
GO:0006974 cellular response to DNA damage stimulus 724 410 91.94 2.71035087719298e-28
GO:0006996 organelle organization 3158 822 401.02 2.71035087719298e-28
GO:0065004 protein-DNA complex assembly 189 145 24 2.71035087719298e-28
GO:0043170 macromolecule metabolic process 7510 1465 953.65 2.71035087719298e-28
GO:0051301 cell division 538 223 68.32 2.71035087719298e-28
GO:0043632 modification-dependent macromolecule cat... 549 331 69.71 2.71035087719298e-28
GO:0036211 protein modification process 3414 902 433.52 2.71035087719298e-28
GO:1901565 organonitrogen compound catabolic proces... 1107 389 140.57 2.71035087719298e-28
GO:0000075 cell cycle checkpoint 204 119 25.9 2.71035087719298e-28
GO:0006260 DNA replication 243 162 30.86 2.71035087719298e-28
GO:0016043 cellular component organization 5129 969 651.3 2.71035087719298e-28
GO:0044265 cellular macromolecule catabolic process 992 387 125.97 2.71035087719298e-28
GO:0009890 negative regulation of biosynthetic proc... 1319 405 167.49 2.71035087719298e-28
GO:0016570 histone modification 403 236 51.17 2.71035087719298e-28
GO:0048523 negative regulation of cellular process 3779 768 479.87 2.71035087719298e-28
GO:0031327 negative regulation of cellular biosynth... 1299 405 164.95 2.71035087719298e-28
GO:0009057 macromolecule catabolic process 1189 409 150.98 2.71035087719298e-28
GO:0031497 chromatin assembly 120 92 15.24 2.71035087719298e-28
GO:0016569 covalent chromatin modification 413 242 52.44 2.71035087719298e-28
GO:0018205 peptidyl-lysine modification 306 159 38.86 2.71035087719298e-28
GO:0006950 response to stress 3200 641 406.35 2.71035087719298e-28
GO:0010558 negative regulation of macromolecule bio... 1234 404 156.7 2.71035087719298e-28
GO:0006281 DNA repair 470 338 59.68 2.71035087719298e-28
GO:0051253 negative regulation of RNA metabolic pro... 1080 352 137.14 2.71035087719298e-28
GO:0043412 macromolecule modification 3579 920 454.48 2.71035087719298e-28
GO:0072331 signal transduction by p53 class mediato... 200 93 25.4 2.71035087719298e-28
GO:0045934 negative regulation of nucleobase-contai... 1206 407 153.14 2.71035087719298e-28
GO:2000113 negative regulation of cellular macromol... 1173 392 148.95 2.71035087719298e-28
GO:0043687 post-translational protein modification 320 184 40.63 2.71035087719298e-28
GO:0040029 regulation of gene expression, epigeneti... 235 124 29.84 2.71035087719298e-28
GO:0006464 cellular protein modification process 3414 902 433.52 2.71035087719298e-28
GO:0048285 organelle fission 401 205 50.92 2.71035087719298e-28
GO:0009889 regulation of biosynthetic process 3399 730 431.62 2.71035087719298e-28
GO:0045892 negative regulation of transcription, DN... 970 336 123.17 4.52815517241379e-28
GO:0009987 cellular process 12542 1688 1592.63 1.004185e-27
GO:0051052 regulation of DNA metabolic process 348 167 44.19 1.46892131147541e-27
GO:0031324 negative regulation of cellular metaboli... 2065 520 262.22 3.48848387096774e-27
GO:0048518 positive regulation of biological proces... 4802 821 609.78 5.069203125e-27
7 GO:0090304 nucleic acid metabolic process 4138 974 525.46 7.13030769230769e-27
GO:0031326 regulation of cellular biosynthetic proc... 3338 727 423.87 6.55412121212121e-26
GO:0198738 cell-cell signaling by wnt 400 104 50.79 8.99270149253731e-26
GO:0006508 proteolysis 1485 456 188.57 3.5823768115942e-25
GO:0019538 protein metabolic process 4819 1010 611.94 5.5175e-25
GO:0010629 negative regulation of gene expression 1472 385 186.92 5.65738028169014e-25
GO:0016458 gene silencing 186 85 23.62 9.01191666666667e-25
GO:0000226 microtubule cytoskeleton organization 436 106 55.37 9.73498630136986e-25
GO:1902679 negative regulation of RNA biosynthetic ... 1009 345 128.13 3.34032432432432e-24
GO:0051983 regulation of chromosome segregation 98 59 12.44 7.62150666666667e-23
GO:0034502 protein localization to chromosome 77 54 9.78 1.01638157894737e-22
GO:1903507 negative regulation of nucleic acid-temp... 1008 345 128 1.18375454545455e-22
GO:0048522 positive regulation of cellular process 4294 765 545.27 2.17870512820513e-22
GO:0009892 negative regulation of metabolic process 2327 534 295.49 3.91113924050633e-22
GO:0045786 negative regulation of cell cycle 495 209 62.86 5.2140375e-22
GO:0045814 negative regulation of gene expression, ... 97 72 12.32 7.91290243902439e-22
GO:0070646 protein modification by small protein re... 258 163 32.76 1.86132530120482e-21
GO:0007275 multicellular organism development 4107 565 521.52 3.86225e-21
GO:0009893 positive regulation of metabolic process 2921 620 370.92 4.18031764705882e-21
GO:1901796 regulation of signal transduction by p53... 120 57 15.24 7.90413953488372e-21
GO:0051172 negative regulation of nitrogen compound... 1946 507 247.11 7.99086206896552e-21
GO:0010556 regulation of macromolecule biosynthetic... 3207 724 407.24 1.931125e-20
GO:0044238 primary metabolic process 8586 1501 1090.29 7.7245e-20
GO:0000079 regulation of cyclin-dependent protein s... 91 53 11.56 1.12047692307692e-19
GO:0036258 multivesicular body assembly 29 28 3.68 1.27622173913043e-19
GO:0019219 regulation of nucleobase-containing comp... 3217 752 408.51 3.48848387096774e-19
GO:0051303 establishment of chromosome localization 67 37 8.51 1.34767872340426e-18
GO:0000003 reproduction 1101 245 139.81 3.86225e-18
GO:0022414 reproductive process 1098 244 139.43 5.57438144329897e-18
GO:0051252 regulation of RNA metabolic process 2978 664 378.16 8.98564285714286e-18
GO:0006355 regulation of transcription, DNA-templat... 2729 641 346.54 1.13916868686869e-17
GO:0045935 positive regulation of nucleobase-contai... 1588 440 201.65 1.220471e-17
GO:0060249 anatomical structure homeostasis 352 92 44.7 1.52960396039604e-17
GO:0050000 chromosome localization 68 37 8.63 2.27191176470588e-17
GO:0009891 positive regulation of biosynthetic proc... 1683 403 213.71 1.46990485436893e-16
GO:1901575 organic substance catabolic process 1828 438 232.13 2,207E-13
GO:1903046 meiotic cell cycle process 157 97 19.94 2,207E-13
GO:0071897 DNA biosynthetic process 166 87 21.08 3.06065094339623e-16
GO:0051254 positive regulation of RNA metabolic pro... 1428 370 181.33 1.03955887850467e-15
GO:0031328 positive regulation of cellular biosynth... 1655 400 210.16 1.43046296296296e-15
GO:0072395 signal transduction involved in cell cyc... 69 39 8.76 1.06300458715596e-14
GO:0044248 cellular catabolic process 1935 460 245.71 1.15165272727273e-14
GO:0031647 regulation of protein stability 239 69 30.35 1.53098198198198e-14
GO:1905114 cell surface receptor signaling pathway ... 482 105 61.21 3.82807079646018e-14
GO:1903513 endoplasmic reticulum to cytosol transpo... 26 20 3.3 4.87863157894737e-14
GO:0060968 regulation of gene silencing 89 45 11.3 5.3735652173913e-14
GO:0000209 protein polyubiquitination 231 183 29.33 5.4604224137931e-14
GO:0009411 response to UV 121 67 15.37 6.20600854700855e-14
GO:0009314 response to radiation 384 134 48.76 1.29823529411765e-13
GO:1904837 beta-catenin-TCF complex assembly 28 25 3.56 1.29823529411765e-13
GO:0071985 multivesicular body sorting pathway 31 19 3.94 1.41615833333333e-13
GO:0051321 meiotic cell cycle 211 115 26.79 1.9151652892562e-13
GO:0033683 nucleotide-excision repair, DNA incision 38 37 4.83 4.55872131147541e-13
GO:0010498 proteasomal protein catabolic process 407 246 51.68 7.47532258064516e-13
GO:0031325 positive regulation of cellular metaboli... 2736 575 347.43 8.280664e-13
GO:0010033 response to organic substance 2704 373 343.37 8.33755555555556e-13
GO:0046755 viral budding 23 23 2.92 1.08264645669291e-12
GO:0006275 regulation of DNA replication 96 59 12.19 1.3276484375e-12
GO:0036257 multivesicular body organization 30 28 3.81 3.47303100775194e-12
GO:0030522 intracellular receptor signaling pathway 257 71 32.63 3.68399230769231e-12
GO:0045787 positive regulation of cell cycle 349 132 44.32 3.77380152671756e-12
GO:0009056 catabolic process 2174 477 276.06 4.99478947368421e-12
GO:0002520 immune system development 756 155 96 4.99478947368421e-12
GO:0010628 positive regulation of gene expression 1621 374 205.84 6.80217164179105e-12
GO:0042176 regulation of protein catabolic process 319 125 40.51 1.25880740740741e-11
7 GO:0098813 nuclear chromosome segregation 227 144 28.83 1.59033823529412e-11
GO:0010557 positive regulation of macromolecule bio... 1575 398 200 2.14256204379562e-11
GO:0051783 regulation of nuclear division 190 101 24.13 2.23898550724638e-11
GO:0009628 response to abiotic stimulus 951 207 120.76 2.88974100719424e-11
GO:0002200 somatic diversification of immune recept... 58 28 7.37 2.97945e-11
GO:2001141 regulation of RNA biosynthetic process 2778 650 352.76 3.28702127659574e-11
GO:0000122 negative regulation of transcription by ... 689 239 87.49 3.78122377622378e-11
GO:0080135 regulation of cellular response to stres... 561 149 71.24 3.86225e-11
GO:0019941 modification-dependent protein catabolic... 539 331 68.44 1.17199310344828e-10
GO:0033365 protein localization to organelle 781 152 99.17 1.26114285714286e-10
GO:0042770 signal transduction in response to DNA d... 123 60 15.62 1.26114285714286e-10
GO:0007032 endosome organization 67 30 8.51 3.11053691275168e-10
GO:0098732 macromolecule deacylation 94 55 11.94 9.2694e-10
GO:0035601 protein deacylation 93 55 11.81 1.51460784313725e-09
GO:0010605 negative regulation of macromolecule met... 2154 521 273.52 2,207E-06
GO:2001022 positive regulation of response to DNA d... 76 41 9.65 3.0898e-09
GO:0010948 negative regulation of cell cycle proces... 256 148 32.51 4.55547435897436e-09
GO:0033036 macromolecule localization 2514 282 319.24 5.70727388535032e-09
GO:0032201 telomere maintenance via semi-conservati... 24 22 3.05 5.86670886075949e-09
GO:0007034 vacuolar transport 118 31 14.98 6.21846540880503e-09
GO:0019068 virion assembly 35 27 4.44 7.5805652173913e-09
GO:0051054 positive regulation of DNA metabolic pro... 186 90 23.62 1.14437037037037e-08
GO:0006293 nucleotide-excision repair, preincision ... 20 20 2.54 1.61124539877301e-08
GO:0051053 negative regulation of DNA metabolic pro... 130 73 16.51 1.68534545454545e-08
GO:0032259 methylation 294 92 37.33 1.68534545454545e-08
GO:0030219 megakaryocyte differentiation 73 38 9.27 1.76825903614458e-08
GO:0007569 cell aging 85 34 10.79 2.65101183431953e-08
GO:0051985 negative regulation of chromosome segreg... 44 29 5.59 2.80069590643275e-08
GO:0051984 positive regulation of chromosome segreg... 26 20 3.3 2.80069590643275e-08
GO:0032527 protein exit from endoplasmic reticulum 39 20 4.95 3.03621965317919e-08
GO:0006342 chromatin silencing 80 59 10.16 3.19634482758621e-08
GO:0008283 cell proliferation 1662 293 211.05 3.26636e-08
GO:0036503 ERAD pathway 79 46 10.03 4.62597175141243e-08
GO:0050789 regulation of biological process 9094 1263 1154.79 6.64565921787709e-08
GO:0030111 regulation of Wnt signaling pathway 262 69 33.27 7.38118888888889e-08
GO:0032204 regulation of telomere maintenance 73 32 9.27 8.27929834254144e-08
GO:0070507 regulation of microtubule cytoskeleton o... 154 50 19.56 8.40357692307692e-08
GO:1904029 regulation of cyclin-dependent protein k... 92 53 11.68 1.18189071038251e-07
GO:0016197 endosomal transport 174 39 22.1 1.25942934782609e-07
GO:0060964 regulation of gene silencing by miRNA 56 26 7.11 1.50314594594595e-07
GO:0009755 hormone-mediated signaling pathway 217 55 27.56 1.65229946524064e-07
GO:0051338 regulation of transferase activity 803 155 101.97 1.65229946524064e-07
GO:0016055 Wnt signaling pathway 398 104 50.54 2.12525925925926e-07
GO:0044786 cell cycle DNA replication 58 46 7.37 2.35800526315789e-07
GO:0033157 regulation of intracellular protein tran... 200 47 25.4 3.55893193717277e-07
GO:1903362 regulation of cellular protein catabolic... 209 84 26.54 3.70132291666667e-07
GO:0051304 chromosome separation 83 60 10.54 3.84223834196891e-07
GO:0044419 interspecies interaction between organis... 737 169 93.59 4.69840721649485e-07
GO:0045637 regulation of myeloid cell differentiati... 199 59 25.27 4.78370050761421e-07
GO:0097193 intrinsic apoptotic signaling pathway 244 61 30.98 4.78370050761421e-07
GO:0051128 regulation of cellular component organiz... 2018 373 256.25 5.22769191919192e-07
GO:0000731 DNA synthesis involved in DNA repair 47 46 5.97 5.66722110552764e-07
GO:0071453 cellular response to oxygen levels 148 44 18.79 7.49730882352941e-07
GO:0045652 regulation of megakaryocyte differentiat... 60 35 7.62 8.99941747572815e-07
GO:0019827 stem cell population maintenance 125 37 15.87 9.70227053140097e-07
GO:0071704 organic substance metabolic process 8885 1508 1128.25 1.03983653846154e-06
GO:0008213 protein alkylation 150 71 19.05 1.1035e-06
GO:0051098 regulation of binding 325 80 41.27 1.1035e-06
GO:0061641 CENP-A containing chromatin organization 34 34 4.32 1.23883490566038e-06
GO:1901564 organonitrogen compound metabolic proces... 5705 1060 724.44 1.4506103286385e-06
GO:0044267 cellular protein metabolic process 4335 988 550.48 1.73259813084112e-06
GO:0042769 DNA damage response, detection of DNA da... 38 29 4.83 1.79639534883721e-06
GO:0032479 regulation of type I interferon producti... 98 26 12.44 1.931125e-06
GO:0051704 multi-organism process 1984 329 251.94 2.1358064516129e-06
7 GO:0051443 positive regulation of ubiquitin-protein... 30 19 3.81 2.46901826484018e-06
GO:0000083 regulation of transcription involved in ... 27 21 3.43 2.46901826484018e-06
GO:0070727 cellular macromolecule localization 1542 208 195.81 2.58648416289593e-06
GO:0032606 type I interferon production 99 26 12.57 2.7836036036036e-06
GO:0032504 multicellular organism reproduction 625 119 79.37 2.84039910313901e-06
GO:0006368 transcription elongation from RNA polyme... 72 38 9.14 3.10359375e-06
GO:0071383 cellular response to steroid hormone sti... 227 56 28.83 3.15846222222222e-06
GO:0010468 regulation of gene expression 3522 689 447.24 3.21284513274336e-06
GO:0010212 response to ionizing radiation 131 70 16.63 3.38793859649123e-06
GO:0015833 peptide transport 1701 169 216 4.2501615720524e-06
GO:1902680 positive regulation of RNA biosynthetic ... 1358 360 172.44 4.36602173913044e-06
GO:0061919 process utilizing autophagic mechanism 392 91 49.78 4.68151515151515e-06
GO:1903706 regulation of hemopoiesis 342 74 43.43 5.52701293103448e-06
GO:0006363 termination of RNA polymerase I transcri... 27 19 3.43 6.63047210300429e-06
GO:0097190 apoptotic signaling pathway 496 96 62.98 7.26235042735043e-06
GO:0051716 cellular response to stimulus 6086 907 772.83 7.88885106382979e-06
GO:0030099 myeloid cell differentiation 336 76 42.67 1.1035e-05
GO:0006998 nuclear envelope organization 49 20 6.22 1.1035e-05
GO:0043543 protein acylation 193 83 24.51 1.34061570247934e-05
GO:0008156 negative regulation of DNA replication 38 30 4.83 1.34061570247934e-05
GO:0006479 protein methylation 150 71 19.05 1.34061570247934e-05
GO:1902750 negative regulation of cell cycle G2/M p... 56 43 7.11 1.65297942386831e-05
GO:1903827 regulation of cellular protein localizat... 435 82 55.24 1.70952049180328e-05
GO:0032886 regulation of microtubule-based process 177 51 22.48 2.05571370967742e-05
GO:0044770 cell cycle phase transition 477 239 60.57 2.05571370967742e-05
GO:0032509 endosome transport via multivesicular bo... 23 16 2.92 2.29563453815261e-05
GO:0022412 cellular process involved in reproductio... 261 65 33.14 2.47184e-05
GO:0006354 DNA-templated transcription, elongation 84 40 10.67 2.57483333333333e-05
GO:1904031 positive regulation of cyclin-dependent ... 33 18 4.19 2.57483333333333e-05
GO:0043414 macromolecule methylation 249 87 31.62 2.62571936758893e-05
GO:0007131 reciprocal meiotic recombination 45 41 5.71 2.67620472440945e-05
GO:0045023 G0 to G1 transition 40 32 5.08 2.96863137254902e-05
GO:0006270 DNA replication initiation 29 28 3.68 3.138078125e-05
GO:0009968 negative regulation of signal transducti... 1022 163 129.78 3.89218992248062e-05
GO:0006473 protein acetylation 170 83 21.59 4.73532567049808e-05
GO:0061458 reproductive system development 343 78 43.56 4.77621755725191e-05
GO:0006338 chromatin remodeling 139 108 17.65 4.87553992395437e-05
GO:0006297 nucleotide-excision repair, DNA gap fill... 23 23 2.92 4.91559090909091e-05
GO:0034728 nucleosome organization 137 108 17.4 5.22710526315789e-05
GO:0035195 gene silencing by miRNA 86 34 10.92 5.22710526315789e-05
GO:0071407 cellular response to organic cyclic comp... 502 99 63.75 5.38111235955056e-05
GO:0031056 regulation of histone modification 123 54 15.62 5.70690671641791e-05
GO:0051383 kinetochore organization 19 16 2.41 5.74312267657993e-05
GO:0002218 activation of innate immune response 206 26 26.16 6.27081180811808e-05
GO:0048608 reproductive structure development 341 77 43.3 6.27081180811808e-05
GO:0045893 positive regulation of transcription, DN... 1279 333 162.41 6.81573529411765e-05
GO:0045815 positive regulation of gene expression, ... 52 29 6.6 7.32981751824818e-05
GO:0098727 maintenance of cell number 128 38 16.25 7.86494545454545e-05
GO:0045732 positive regulation of protein catabolic... 183 74 23.24 8.39619565217391e-05
GO:0000070 mitotic sister chromatid segregation 142 89 18.03 9.4813357400722e-05
GO:0034508 centromere complex assembly 46 42 5.84 0.000105586690647482
GO:0032922 circadian regulation of gene expression 51 20 6.48 0.000110745519713262
GO:0010639 negative regulation of organelle organiz... 327 118 41.52 0.000115455160142349
GO:0007276 gamete generation 507 110 64.38 0.000120524113475177
GO:0061726 mitochondrion disassembly 64 21 8.13 0.00012511514084507
GO:0070316 regulation of G0 to G1 transition 38 30 4.83 0.000140938245614035
GO:0034724 DNA replication-independent nucleosome o... 45 44 5.71 0.000145847202797203
GO:0003006 developmental process involved in reprod... 518 112 65.78 0.000150721951219512
GO:0010604 positive regulation of macromolecule met... 2751 607 349.33 0.000171655555555556
GO:0061418 regulation of transcription from RNA pol... 27 19 3.43 0.000176407266435986
7 GO:0033143 regulation of intracellular steroid horm... 72 24 9.14 0.000232793150684932
GO:0030177 positive regulation of Wnt signaling pat... 109 32 13.84 0.000232793150684932
GO:0002377 immunoglobulin production 80 23 10.16 0.000257483333333333
GO:0071705 nitrogen compound transport 1944 176 246.86 0.000257483333333333
GO:0009790 embryo development 772 146 98.03 0.000272321355932203
GO:0051656 establishment of organelle localization 398 57 50.54 0.000292278378378378
GO:0016032 viral process 631 163 80.13 0.000321422147651007
GO:0035825 homologous recombination 45 41 5.71 0.000410604651162791
GO:0007088 regulation of mitotic nuclear division 167 93 21.21 0.000418091749174917
GO:0061982 meiosis I cell cycle process 95 72 12.06 0.000418091749174917
GO:0034622 cellular protein-containing complex asse... 939 214 119.24 0.000467535526315789
GO:0009894 regulation of catabolic process 732 182 92.95 0.000484674509803922
GO:0090169 regulation of spindle assembly 25 16 3.17 0.000548190322580645
GO:0006352 DNA-templated transcription, initiation 211 75 26.79 0.000548190322580645
GO:0040007 growth 764 137 97.02 0.000596102893890675
GO:0000715 nucleotide-excision repair, DNA damage r... 22 22 2.79 0.000643708333333333
GO:1903508 positive regulation of nucleic acid-temp... 1357 360 172.32 0.000738009554140127
GO:0002381 immunoglobulin production involved in im... 44 18 5.59 0.000738009554140127
GO:0007389 pattern specification process 349 65 44.32 0.000828495268138801
GO:0061025 membrane fusion 148 29 18.79 0.000828495268138801
GO:0033045 regulation of sister chromatid segregati... 77 51 9.78 0.000828495268138801
GO:0048511 rhythmic process 247 55 31.37 0.000871730407523511
GO:0007281 germ cell development 192 46 24.38 0.000914426791277259
GO:0032386 regulation of intracellular transport 324 50 41.14 0.000914426791277259
GO:0006333 chromatin assembly or disassembly 144 108 18.29 0.000956594427244582
GO:0090342 regulation of cell aging 37 16 4.7 0.000956594427244582
GO:0006753 nucleoside phosphate metabolic process 523 84 12.84 5.14966666666667e-27
GO:0098781 ncRNA transcription 90 37 2.21 5.14966666666667e-27
GO:0042795 snRNA transcription by RNA polymerase II 66 32 1.62 5.14966666666667e-27
GO:0009116 nucleoside metabolic process 109 37 2.68 1.08143e-26
GO:0055086 nucleobase-containing small molecule met... 597 89 14.66 3.70776e-26
GO:1901293 nucleoside phosphate biosynthetic proces... 284 59 6.97 1.58904e-19
GO:0043094 cellular metabolic compound salvage 34 19 0.83 3.60476666666667e-17
8 GO:0019693 ribose phosphate metabolic process 413 54 10.14 7.87899e-17
GO:0072527 pyrimidine-containing compound metabolic... 80 26 1.96 1.2745425e-16
GO:0019637 organophosphate metabolic process 965 84 23.69 1.66373846153846e-16
GO:0072521 purine-containing compound metabolic pro... 432 55 10.61 4.22272666666667e-16
GO:0016073 snRNA metabolic process 83 32 2.04 5.11748125e-15
GO:0009163 nucleoside biosynthetic process 39 21 0.96 2.23152222222222e-14
GO:0006383 transcription by RNA polymerase III 45 19 1.1 1.05703684210526e-13
GO:0034404 nucleobase-containing small molecule bio... 157 37 3.85 1.5449e-13
GO:1901657 glycosyl compound metabolic process 130 37 3.19 3.51113636363636e-13
GO:0072522 purine-containing compound biosynthetic ... 228 41 5.6 3.51113636363636e-13
GO:0090407 organophosphate biosynthetic process 569 62 13.97 2.38172083333333e-11
GO:1901135 carbohydrate derivative metabolic proces... 999 74 24.53 1.820775e-10
GO:1901659 glycosyl compound biosynthetic process 42 21 1.03 4.6879724137931e-10
GO:0072528 pyrimidine-containing compound biosynthe... 39 16 0.96 6.97696774193548e-10
8 GO:0006457 protein folding 191 26 4.69 8.6900625e-10
GO:0044281 small molecule metabolic process 1779 103 43.68 4.26017878787879e-09
GO:0046390 ribose phosphate biosynthetic process 226 41 5.55 4,414E-05
GO:0034660 ncRNA metabolic process 464 43 11.39 6.00794444444444e-08
GO:1901137 carbohydrate derivative biosynthetic pro... 628 54 15.42 3.61572340425532e-06
GO:0034656 nucleobase-containing small molecule cat... 44 16 1.08 2.55381428571429e-05
GO:0034654 nucleobase-containing compound biosynthe... 3345 182 82.12 0.000210668181818182
GO:1901566 organonitrogen compound biosynthetic pro... 1636 83 40.17 0.000825722413793103
GO:0051169 nuclear transport 323 57 5.58 5.87062e-25
GO:0017038 protein import 192 34 3.32 3.0898e-15
GO:0016071 mRNA metabolic process 692 62 11.96 2.26585333333333e-13
GO:0006397 mRNA processing 439 58 7.59 6.79756e-11
GO:0006396 RNA processing 781 62 13.5 6.79756e-11
GO:0051236 establishment of RNA localization 177 24 3.06 9.2694e-11
GO:0006457 protein folding 191 22 3.3 1.75732375e-09
GO:0006606 protein import into nucleus 139 30 2.4 1.63072777777778e-08
GO:0015931 nucleobase-containing compound transport 201 24 3.47 2.16286e-08
GO:0006353 DNA-templated transcription, termination 68 16 1.18 2.24712727272727e-08
GO:0006403 RNA localization 208 24 3.59 2.75875e-07
GO:0010467 gene expression 4333 135 74.88 2.80013125e-07
9 GO:0008380 RNA splicing 378 53 6.53 2.80013125e-07
GO:0071826 ribonucleoprotein complex subunit organi... 224 22 3.87 8.94415789473684e-07
GO:0034504 protein localization to nucleus 233 33 4.03 1.313165e-06
GO:0022618 ribonucleoprotein complex assembly 213 22 3.68 4.11973333333333e-06
GO:0072594 establishment of protein localization to... 500 38 8.64 6.79756e-06
GO:0006406 mRNA export from nucleus 102 17 1.76 7.7245e-06
GO:0071166 ribonucleoprotein complex localization 122 18 2.11 1.97107931034483e-05
GO:0000387 spliceosomal snRNP assembly 37 17 0.64 6.69456666666667e-05
GO:1901360 organic cyclic compound metabolic proces... 5009 131 86.57 0.000561781818181818
GO:0015833 peptide transport 1701 57 29.4 0.000590697058823529
GO:0006518 peptide metabolic process 763 214 29.26 5.94192307692308e-28
GO:0022613 ribonucleoprotein complex biogenesis 394 173 15.11 5.94192307692308e-28
GO:0006412 translation 626 212 24.01 5.94192307692308e-28
GO:0043604 amide biosynthetic process 713 212 27.34 5.94192307692308e-28
GO:0006413 translational initiation 185 115 7.09 5.94192307692308e-28
GO:0043043 peptide biosynthetic process 644 212 24.7 5.94192307692308e-28
GO:1901566 organonitrogen compound biosynthetic pro... 1636 224 62.74 5.94192307692308e-28
GO:0034660 ncRNA metabolic process 464 169 17.79 5.94192307692308e-28
10 GO:0006401 RNA catabolic process 304 132 11.66 5.94192307692308e-28
GO:0006402 mRNA catabolic process 274 129 10.51 5.94192307692308e-28
GO:0043603 cellular amide metabolic process 902 214 34.59 5.94192307692308e-28
GO:0010467 gene expression 4333 406 166.16 5.94192307692308e-28
GO:0046700 heterocycle catabolic process 526 140 20.17 5.94192307692308e-28
GO:0019439 aromatic compound catabolic process 540 140 20.71 5.94192307692308e-28
GO:1901361 organic cyclic compound catabolic proces... 570 140 21.86 5.94192307692308e-28
GO:0034655 nucleobase-containing compound catabolic... 479 138 18.37 5.94192307692308e-28
GO:0044270 cellular nitrogen compound catabolic pro... 526 140 20.17 5.94192307692308e-28
GO:0070972 protein localization to endoplasmic reti... 128 69 4.91 5.94192307692308e-28
GO:0045047 protein targeting to ER 104 69 3.99 5.94192307692308e-28
GO:0071826 ribonucleoprotein complex subunit organi... 224 68 8.59 5.94192307692308e-28
GO:0016071 mRNA metabolic process 692 156 26.54 5.94192307692308e-28
GO:0043038 amino acid activation 48 32 1.84 5.94192307692308e-28
GO:0044265 cellular macromolecule catabolic process 992 137 38.04 5.94192307692308e-28
GO:0010608 posttranscriptional regulation of gene e... 422 97 16.18 2.45053103448276e-26
GO:0034645 cellular macromolecule biosynthetic proc... 3932 299 150.78 4.17123e-26
GO:0006612 protein targeting to membrane 166 69 6.37 1.5449e-25
GO:0043170 macromolecule metabolic process 7510 441 287.99 7.7245e-23
GO:0034248 regulation of cellular amide metabolic p... 381 78 14.61 7.5038e-22
GO:0006417 regulation of translation 341 77 13.08 1.67016216216216e-21
GO:0006613 cotranslational protein targeting to mem... 98 69 3.76 4.47207894736842e-21
GO:0006396 RNA processing 781 153 29.95 5.54579487179487e-21
GO:0033036 macromolecule localization 2514 141 96.41 5.5175e-19
GO:0006886 intracellular protein transport 958 101 36.74 2.11974651162791e-18
GO:0009892 negative regulation of metabolic process 2327 197 89.23 4.80635555555556e-18
GO:0071705 nitrogen compound transport 1944 116 74.55 1.27622173913043e-17
GO:0044085 cellular component biogenesis 2670 207 102.39 2.49813617021277e-17
GO:0009057 macromolecule catabolic process 1189 138 45.6 1.31960208333333e-16
GO:0034470 ncRNA processing 312 121 11.96 6.1796e-16
GO:0045184 establishment of protein localization 1760 112 67.49 6.1796e-16
10 GO:0006364 rRNA processing 174 107 6.67 9.50707692307692e-15
GO:0000956 nuclear-transcribed mRNA catabolic proce... 190 120 7.29 1.51575094339623e-14
GO:0072331 signal transduction by p53 class mediato... 200 26 7.67 4.29138888888889e-14
GO:0090501 RNA phosphodiester bond hydrolysis 127 44 4.87 4.77514545454545e-14
GO:0070727 cellular macromolecule localization 1542 119 59.13 2.46641929824561e-13
GO:0071702 organic substance transport 2254 117 86.44 3.86225e-12
GO:0051236 establishment of RNA localization 177 31 6.79 5.31850819672131e-12
GO:0043487 regulation of RNA stability 105 28 4.03 5.98025806451613e-12
GO:0016072 rRNA metabolic process 201 112 7.71 8.33755555555556e-12
GO:0006403 RNA localization 208 41 7.98 1.206953125e-11
GO:0006605 protein targeting 371 74 14.23 2.32923384615385e-11
GO:0016458 gene silencing 186 33 7.13 9.3630303030303e-11
GO:0090305 nucleic acid phosphodiester bond hydroly... 241 55 9.24 1.5449e-10
GO:0072657 protein localization to membrane 483 72 18.52 1.97656323529412e-10
GO:0034622 cellular protein-containing complex asse... 939 87 36.01 2.23898550724638e-10
GO:0010605 negative regulation of macromolecule met... 2154 193 82.6 3.0898e-10
GO:0061013 regulation of mRNA catabolic process 119 32 4.56 7.40705479452055e-10
GO:0043039 tRNA aminoacylation 47 32 1.8 5.14966666666667e-09
GO:0043933 protein-containing complex subunit organ... 1878 110 72.02 1.38439090909091e-08
GO:0042254 ribosome biogenesis 241 136 9.24 3.16902564102564e-08
GO:0042255 ribosome assembly 54 30 2.07 6.40568292682927e-08
GO:1901575 organic substance catabolic process 1828 145 70.1 7.07303614457831e-08
GO:0015931 nucleobase-containing compound transport 201 32 7.71 2.87423255813953e-07
GO:0071426 ribonucleoprotein complex export from nu... 121 24 4.64 4.74003409090909e-07
GO:0002181 cytoplasmic translation 88 55 3.37 4.74003409090909e-07
GO:0022618 ribonucleoprotein complex assembly 213 67 8.17 1.05886404494382e-06
GO:0071166 ribonucleoprotein complex localization 122 24 4.68 1.39210769230769e-06
GO:0015833 peptide transport 1701 111 65.23 1.84716304347826e-06
GO:0044248 cellular catabolic process 1935 149 74.2 2.6296170212766e-06
GO:0034613 cellular protein localization 1533 115 58.79 2.6296170212766e-06
GO:1903311 regulation of mRNA metabolic process 227 39 8.7 2.8966875e-06
GO:0065003 protein-containing complex assembly 1601 96 61.39 3.62578571428571e-06
GO:0016070 RNA metabolic process 3693 333 141.62 1.07993009708738e-05
GO:0010629 negative regulation of gene expression 1472 175 56.45 1.12896538461538e-05
GO:0033365 protein localization to organelle 781 85 29.95 2.14569444444444e-05
GO:0042273 ribosomal large subunit biogenesis 63 46 2.42 4.78508849557522e-05
10 GO:0009451 RNA modification 128 24 4.91 5.23922608695652e-05
GO:0043414 macromolecule methylation 249 34 9.55 9.68835593220339e-05
GO:0006418 tRNA aminoacylation for protein translat... 44 30 1.69 0.00019151652892562
GO:0070925 organelle assembly 670 50 25.69 0.000238643089430894
GO:0044238 primary metabolic process 8586 444 329.25 0.000299012903225806
GO:0009056 catabolic process 2174 149 83.37 0.0003213392
GO:0032259 methylation 294 34 11.27 0.00033105
GO:0090503 RNA phosphodiester bond hydrolysis, exon... 37 24 1.42 0.0008931453125
GO:0042274 ribosomal small subunit biogenesis 58 41 2.22 0.000958077519379845
GO:0006119 oxidative phosphorylation 113 38 1.22 5.14966666666667e-27
GO:0006091 generation of precursor metabolites and ... 412 47 4.46 5.14966666666667e-27
GO:0072521 purine-containing compound metabolic pro... 432 43 4.68 5.14966666666667e-27
GO:0019693 ribose phosphate metabolic process 413 43 4.47 5.793375e-26
GO:0007005 mitochondrion organization 437 35 4.73 8.34246e-21
GO:0006753 nucleoside phosphate metabolic process 523 44 5.66 9.01191666666667e-21
GO:0055086 nucleobase-containing small molecule met... 597 44 6.47 1.80974e-20
GO:0046034 ATP metabolic process 220 43 2.38 6.7589375e-20
GO:0017144 drug metabolic process 694 45 7.52 1.88821111111111e-19
GO:1902600 proton transmembrane transport 134 38 1.45 1.096879e-18
GO:0022900 electron transport chain 166 38 1.8 1.68534545454545e-16
GO:0019637 organophosphate metabolic process 965 44 10.45 2.83231666666667e-15
11 GO:0055114 oxidation-reduction process 863 44 9.35 3.80283076923077e-15
GO:1901135 carbohydrate derivative metabolic proces... 999 44 10.82 1.1035e-14
GO:0009123 nucleoside monophosphate metabolic proce... 282 44 3.05 2.98680666666667e-10
GO:0009141 nucleoside triphosphate metabolic proces... 273 43 2.96 2.8966875e-09
GO:0044281 small molecule metabolic process 1779 51 19.27 4.6347e-08
GO:0055085 transmembrane transport 1229 48 13.31 4.6347e-06
GO:0015980 energy derivation by oxidation of organi... 232 31 2.51 2.6484e-05
GO:0015672 monovalent inorganic cation transport 439 41 4.75 7.7245e-05
GO:0034220 ion transmembrane transport 918 47 9.94 0.00015449
GO:0045333 cellular respiration 153 31 1.66 0.0004758292
GO:0006163 purine nucleotide metabolic process 401 43 4.34 0.000588250384615385
Table fields:
Cluster - Cluster number on first time-interval

GO.ID - Gene Ontology term ID
Term - Human-readable term name
Annotated - Number of genes annotated for a specific GO term
Significant - Number of differentially expressed genes identified
Expected - Number of differentially expressed genes expected by the topGO statistical test
pValue - p-values adjusted by the Benjamini-Hochberg procedure.
GO:0097503 sialylation 19 19 4,48 1,65794146341463E-14
GO:1901685 glutathione derivative metabolic process 20 20 4,71 5,793375E-13
GO:1901687 glutathione derivative biosynthetic proc... 20 20 4,71 1,39635192307692E-11
GO:0006506 GPI anchor biosynthetic process 26 26 6,13 9,62396721311475E-11
GO:0006505 GPI anchor metabolic process 27 27 6,36 1,47359692307692E-10
GO:0036149 phosphatidylinositol acyl-chain remodeli... 14 14 3,3 2,51495348837209E-07
GO:1901750 leukotriene D4 biosynthetic process 7 7 1,65 4,16877777777778E-05
GO:0015858 nucleoside transport 7 7 1,65 8,4910534351145E-05
GO:0052646 alditol phosphate metabolic process 8 8 1,89 0,000302497902098
GO:0002084 protein depalmitoylation 4 4 0,94 0,000445843624161
GO:1901748 leukotriene D4 metabolic process 7 7 1,65 0,000738442767296
GO:0016255 attachment of GPI anchor to protein 6 6 1,41 0,000848741358025
GO:0030311 poly-N-acetyllactosamine biosynthetic pr... 8 8 1,89 0,000942012195122
GO:0006829 zinc ion transport 27 26 6,36 6,46702325581395E-14
GO:0071577 zinc ion transmembrane transport 22 21 5,19 5,3403950617284E-08
GO:0006004 fucose metabolic process 14 13 3,3 6,94337078651685E-07
GO:0018146 keratan sulfate biosynthetic process 27 25 6,36 1,79935411764706E-07
GO:0009247 glycolipid biosynthetic process 55 50 12,96 8,53760526315789E-15
GO:0042339 keratan sulfate metabolic process 32 29 7,54 8,64096610169492E-11
GO:0016338 calcium-independent cell-cell adhesion v... 21 19 4,95 0,000161407462687
GO:0042738 exogenous drug catabolic process 20 18 4,71 6,25316666666667E-08
GO:0019755 one-carbon compound transport 10 9 2,36 0,000473769333333
GO:0046473 phosphatidic acid metabolic process 38 34 8,96 5,95624096385542E-08
GO:0006654 phosphatidic acid biosynthetic process 37 33 8,72 1,36232090909091E-05
GO:0046471 phosphatidylglycerol metabolic process 25 22 5,89 0,000201508695652
GO:0006658 phosphatidylserine metabolic process 24 21 5,66 0,000180426277372
GO:0009804 coumarin metabolic process 8 7 1,89 0,000647861290323
GO:0006063 uronic acid metabolic process 23 20 5,42 1,36232090909091E-05
GO:0009812 flavonoid metabolic process 14 12 3,3 3,73766129032258E-05
GO:1903509 liposaccharide metabolic process 90 77 21,21 5,14966666666667E-22
GO:0016137 glycoside metabolic process 18 15 4,24 1,52283E-09
GO:0031424 keratinization 94 77 22,16 9,08764705882353E-28
GO:0006569 tryptophan catabolic process 10 8 2,36 0,000386225
GO:0042436 indole-containing compound catabolic pro... 10 8 2,36 0,000487863157895
GO:0070268 cornification 69 55 16,26 9,08764705882353E-28
A GO:0009311 oligosaccharide metabolic process 47 37 11,08 1,28741666666667E-08
GO:0046470 phosphatidylcholine metabolic process 63 49 14,85 2,657228E-21
GO:0006656 phosphatidylcholine biosynthetic process 26 20 6,13 2,13656382978723E-06
GO:0006805 xenobiotic metabolic process 102 77 24,04 9,08764705882353E-28
GO:0006882 cellular zinc ion homeostasis 32 24 7,54 1,3759265625E-10
GO:0030149 sphingolipid catabolic process 23 17 5,42 2,54828865979381E-06
GO:0046467 membrane lipid biosynthetic process 126 93 29,7 9,08764705882353E-28
GO:0006643 membrane lipid metabolic process 175 129 41,25 8,71482051282051E-15
GO:0015669 gas transport 19 14 4,48 0,000311125694444
GO:0006690 icosanoid metabolic process 91 65 21,45 1,303509375E-17
GO:0042430 indole-containing compound metabolic pro... 21 15 4,95 3,64244715447154E-05
GO:0097501 stress response to metal ion 14 10 3,3 0,000494771895425
GO:0055069 zinc ion homeostasis 34 24 8,01 3,43311111111111E-09
GO:0006749 glutathione metabolic process 44 31 10,37 1,83099259259259E-11
GO:0018149 peptide cross-linking 47 33 11,08 1,5449E-15
GO:0034308 primary alcohol metabolic process 47 33 11,08 0,000581845454545
GO:0001867 complement activation, lectin pathway 13 9 3,06 0,000723560759494
GO:0001676 long-chain fatty acid metabolic process 92 63 21,69 9,71791935483871E-11
GO:0006672 ceramide metabolic process 79 54 18,62 2,84586842105263E-23
GO:0046456 icosanoid biosynthetic process 41 28 9,66 3,954944E-05
GO:0006665 sphingolipid metabolic process 133 89 31,35 9,08764705882353E-28
GO:0072348 sulfur compound transport 33 22 7,78 9,81805607476636E-06
GO:0009310 amine catabolic process 21 14 4,95 1,99771551724138E-05
GO:0030148 sphingolipid biosynthetic process 90 59 21,21 4,54382352941177E-16
GO:0033559 unsaturated fatty acid metabolic process 87 57 20,51 5,46367073170732E-08
GO:0034754 cellular hormone metabolic process 100 65 23,57 1,14437037037037E-20
GO:0070085 glycosylation 222 143 52,33 9,08764705882353E-28
GO:0043413 macromolecule glycosylation 215 137 50,68 9,08764705882353E-28
GO:0006486 protein glycosylation 215 137 50,68 9,08764705882353E-28
GO:0042537 benzene-containing compound metabolic pr... 22 14 5,19 5,14966666666667E-05
GO:0009074 aromatic amino acid family catabolic pro... 19 12 4,48 2,57483333333333E-05
GO:0007271 synaptic transmission, cholinergic 27 17 6,36 1,99771551724138E-05
GO:0098661 inorganic anion transmembrane transport 91 57 21,45 1,31602592592593E-05
GO:0046513 ceramide biosynthetic process 48 30 11,31 2,30091489361702E-13
GO:0009072 aromatic amino acid family metabolic pro... 29 18 6,84 2,43931578947368E-06
GO:1901568 fatty acid derivative metabolic process 135 82 31,82 4,83620869565217E-22
GO:0030216 keratinocyte differentiation 157 95 37,01 1,59583076923077E-06
GO:0006497 protein lipidation 63 38 14,85 1,0428075E-14
GO:0097164 ammonium ion metabolic process 174 101 41,02 9,08764705882353E-28
GO:1901570 fatty acid derivative biosynthetic proce... 78 45 18,39 1,22611111111111E-10
GO:0042158 lipoprotein biosynthetic process 68 39 16,03 1,82579090909091E-10
GO:0019184 nonribosomal peptide biosynthetic proces... 14 8 3,3 4,28289108910891E-06
GO:0071466 cellular response to xenobiotic stimulus 147 83 34,65 9,08764705882353E-28
GO:1901264 carbohydrate derivative transport 52 29 12,26 2,13656382978723E-06
GO:0098656 anion transmembrane transport 206 114 48,56 3,55149425287356E-07
GO:0042157 lipoprotein metabolic process 95 52 22,39 2,00419459459459E-15
GO:0009913 epidermal cell differentiation 201 110 47,38 7,9452E-16
GO:0043171 peptide catabolic process 24 13 5,66 0,000688808917197
GO:0021522 spinal cord motor neuron differentiation 26 14 6,13 0,000844417391304
GO:0010043 response to zinc ion 43 23 10,14 0,000942012195122
GO:1901616 organic hydroxy compound catabolic proce... 65 34 15,32 3,39878E-06
GO:0042446 hormone biosynthetic process 73 38 17,21 6,49560227272727E-07
GO:0019748 secondary metabolic process 50 26 11,79 0,000108026842105
GO:0009101 glycoprotein biosynthetic process 282 146 66,47 9,08764705882353E-28
GO:0008654 phospholipid biosynthetic process 240 120 56,57 1,776635E-22
GO:0045104 intermediate filament cytoskeleton organ... 40 20 9,43 3,27706060606061E-06
GO:0042445 hormone metabolic process 190 94 44,79 1,91942121212121E-16
GO:0045017 glycerolipid biosynthetic process 232 114 54,69 1,9863E-20
GO:0017001 antibiotic catabolic process 51 25 12,02 5,99961165048544E-06
GO:0009100 glycoprotein metabolic process 342 167 80,62 9,08764705882353E-28
GO:0098754 detoxification 101 49 23,81 5,60404901960784E-06
GO:0051187 cofactor catabolic process 58 28 13,67 9,76493396226415E-06
GO:0006820 anion transport 448 212 105,6 2,54224050632911E-08
GO:0071804 cellular potassium ion transport 179 84 42,19 1,93814727272727E-11
GO:0071805 potassium ion transmembrane transport 179 84 42,19 2,11268376068376E-05
GO:0042737 drug catabolic process 127 59 29,94 1,44190666666667E-13
GO:0015711 organic anion transport 338 157 79,67 3,77642222222222E-26
GO:0043588 skin development 257 119 60,58 1,43835517241379E-19
GO:0016042 lipid catabolic process 275 127 64,82 4,01674E-12
GO:0006956 complement activation 63 29 14,85 4,50088976377953E-05
GO:0008610 lipid biosynthetic process 618 283 145,68 9,08764705882353E-28
GO:0015748 organophosphate ester transport 66 30 15,56 0,000293748591549
A GO:0044255 cellular lipid metabolic process 927 419 218,51 9,08764705882353E-28
GO:0006026 aminoglycan catabolic process 60 27 14,14 1,66244673913043E-06
GO:1990748 cellular detoxification 87 39 20,51 0,000171655555556
GO:0008544 epidermis development 294 131 69,3 1,89374838709677E-18
GO:0015893 drug transport 171 76 40,31 9,88736E-09
GO:0044272 sulfur compound biosynthetic process 166 72 39,13 1,32100144927536E-09
GO:0006575 cellular modified amino acid metabolic p... 158 68 37,24 1,08343636363636E-08
GO:0080171 lytic vacuole organization 49 21 11,55 1,5173125E-05
GO:0006629 lipid metabolic process 1205 510 284,04 9,08764705882353E-28
GO:0098742 cell-cell adhesion via plasma-membrane a... 166 70 39,13 8,40607352941176E-10
GO:0018958 phenol-containing compound metabolic pro... 83 35 19,56 2,54828865979381E-06
GO:0015849 organic acid transport 242 101 57,04 1,81357826086957E-13
GO:0006022 aminoglycan metabolic process 150 62 35,36 1,01638157894737E-08
GO:0006644 phospholipid metabolic process 398 164 93,82 1,14437037037037E-20
GO:0008202 steroid metabolic process 281 114 66,24 1,01822954545455E-13
GO:0006066 alcohol metabolic process 296 119 69,77 7,42740384615385E-06
GO:0009410 response to xenobiotic stimulus 236 94 55,63 2,33311428571429E-12
GO:1901615 organic hydroxy compound metabolic proce... 445 177 104,9 2,57483333333333E-19
GO:0009308 amine metabolic process 78 31 18,39 0,0008110725
GO:0044242 cellular lipid catabolic process 180 71 42,43 0,000336304761905
GO:0002920 regulation of humoral immune response 61 24 14,38 0,000480862913907
GO:0006869 lipid transport 275 106 64,82 3,3105E-11
GO:0010876 lipid localization 300 113 70,72 3,01957727272727E-22
GO:0044106 cellular amine metabolic process 70 26 16,5 0,000233402158273
GO:0006790 sulfur compound metabolic process 323 118 76,14 5,32724137931035E-11
GO:0055085 transmembrane transport 1229 438 289,7 9,08764705882353E-28
GO:0006811 ion transport 1299 462 306,2 9,08764705882353E-28
GO:0006694 steroid biosynthetic process 169 60 39,84 0,000683321153846
GO:0005975 carbohydrate metabolic process 518 177 122,1 9,08764705882353E-12
GO:0032787 monocarboxylic acid metabolic process 511 174 120,45 2,207E-05
GO:1901137 carbohydrate derivative biosynthetic pro... 628 212 148,03 5,14966666666667E-14
GO:0030855 epithelial cell differentiation 526 168 123,99 4,1504776119403E-10
GO:0009593 detection of chemical stimulus 424 132 99,95 3,6691375E-08
GO:0006082 organic acid metabolic process 968 289 228,18 4,60759649122807E-11
GO:1901135 carbohydrate derivative metabolic proces... 999 297 235,49 1,51575094339623E-11
GO:0055114 oxidation-reduction process 863 256 203,43 7,93327027027027E-09
GO:0050877 nervous system process 1100 320 259,29 0,000317445205479
GO:0051606 detection of stimulus 577 167 136,01 1,76172807017544E-05
GO:0051186 cofactor metabolic process 496 141 116,92 5,069203125E-05
GO:0010951 negative regulation of endopeptidase act... 187 51 44,08 0,000273918439716
GO:0010466 negative regulation of peptidase activit... 191 51 45,02 8,43753076923077E-05
GO:0044281 small molecule metabolic process 1779 475 419,35 1,7407323943662E-09
GO:0051259 protein complex oligomerization 445 115 104,9 2,13343333333333E-22
GO:0007186 G-protein coupled receptor signaling pat... 1174 256 276,74 1,37324444444444E-06
GO:0051346 negative regulation of hydrolase activit... 342 73 80,62 2,80890909090909E-05
GO:0044703 multi-organism reproductive process 749 151 176,56 0,00024277
A GO:0006357 regulation of transcription by RNA polym... 2066 401 487 7,93327027027027E-09
GO:0006952 defense response 1288 242 303,61 0,000180426277372
GO:0006366 transcription by RNA polymerase II 2189 403 515,99 1,5173125E-05
GO:2001141 regulation of RNA biosynthetic process 2778 480 654,83 1,76172807017544E-05
GO:0006355 regulation of transcription, DNA-templat... 2729 471 643,28 2,207E-05
GO:1903506 regulation of nucleic acid-templated tra... 2774 478 653,89 3,54567213114754E-05
GO:0043086 negative regulation of catalytic activit... 620 105 146,15 0,000317445205479
GO:0051252 regulation of RNA metabolic process 2978 496 701,98 9,41653333333333E-06
GO:0010468 regulation of gene expression 3522 560 830,21 8,54376515151515E-05
GO:0016070 RNA metabolic process 3693 519 870,52 9,2694E-11
GO:0007010 cytoskeleton organization 1076 147 253,64 2,99521428571429E-06
GO:0030208 dermatan sulfate biosynthetic process 12 12 1,92 1,58665405405405E-06
GO:0072378 blood coagulation, fibrin clot formation 26 25 4,15 7,93906944444444E-08
GO:0030206 chondroitin sulfate biosynthetic process 23 22 3,67 2,3509347826087E-08
GO:0030205 dermatan sulfate metabolic process 13 12 2,08 6,95656725146199E-07
GO:0007200 phospholipase C-activating G-protein cou... 86 78 13,73 1,14823648648649E-07
GO:0018298 protein-chromophore linkage 10 9 1,6 5,09893103448276E-06
GO:0051918 negative regulation of fibrinolysis 10 9 1,6 3,41081818181818E-05
GO:0006768 biotin metabolic process 7 6 1,12 0,000601909090909
GO:0006171 cAMP biosynthetic process 40 34 6,38 3,25242105263158E-22
GO:1902285 semaphorin-plexin signaling pathway invo... 13 11 2,08 0,000403017391304
GO:0035815 positive regulation of renal sodium excr... 12 10 1,92 0,000506440728477
GO:0030049 muscle filament sliding 34 28 5,43 0,000384924579125
GO:0030814 regulation of cAMP metabolic process 28 23 4,47 4,28104819277108E-07
GO:0030817 regulation of cAMP biosynthetic process 28 23 4,47 2,482875E-05
GO:0071377 cellular response to glucagon stimulus 22 18 3,51 4,27045528455285E-09
GO:0022401 negative adaptation of signaling pathway 15 12 2,39 1,30554929577465E-05
GO:0071545 inositol phosphate catabolic process 14 11 2,23 0,000102191828794
GO:0007190 activation of adenylate cyclase activity 27 21 4,31 4,81046706586826E-07
GO:0002430 complement receptor mediated signaling p... 9 7 1,44 0,00055535620915
GO:0030166 proteoglycan biosynthetic process 56 42 8,94 2,78753695652174E-18
GO:0035587 purinergic receptor signaling pathway 28 21 4,47 1,81752941176471E-08
GO:0007210 serotonin receptor signaling pathway 36 27 5,75 1,43235761589404E-07
GO:1902224 ketone body metabolic process 12 9 1,92 0,000245655477032
GO:0002576 platelet degranulation 119 88 18,99 1,489725E-26
4 GO:0031280 negative regulation of cyclase activity 19 14 3,03 0,000201508695652
GO:0070098 chemokine-mediated signaling pathway 75 55 11,97 5,94192307692308E-28
GO:0048520 positive regulation of behavior 22 16 3,51 1,04150561797753E-06
GO:0046950 cellular ketone body metabolic process 11 8 1,76 0,000171655555556
GO:0046058 cAMP metabolic process 50 36 7,98 1,5449E-21
GO:0046856 phosphatidylinositol dephosphorylation 25 18 3,99 4,28104819277108E-07
GO:0030799 regulation of cyclic nucleotide metaboli... 39 28 6,23 7,07587786259542E-09
GO:0030802 regulation of cyclic nucleotide biosynth... 39 28 6,23 1,95145263157895E-06
GO:0030204 chondroitin sulfate metabolic process 35 25 5,59 2,00837E-10
GO:0032958 inositol phosphate biosynthetic process 24 17 3,83 4,17028220858896E-07
GO:1900048 positive regulation of hemostasis 24 17 3,83 7,23334682080925E-07
GO:0006560 proline metabolic process 10 7 1,6 3,35847826086957E-05
GO:0031034 myosin filament assembly 10 7 1,6 4,12409745762712E-05
GO:0050819 negative regulation of coagulation 50 34 7,98 1,33891333333333E-07
GO:0050820 positive regulation of coagulation 25 17 3,99 0,000201508695652
GO:0031279 regulation of cyclase activity 43 29 6,86 1,51460784313725E-07
GO:1900047 negative regulation of hemostasis 48 32 7,66 6,82163636363636E-13
GO:0051339 regulation of lyase activity 42 28 6,7 4,23905487804878E-07
GO:1905144 response to acetylcholine 24 16 3,83 7,47532258064516E-05
GO:1905145 cellular response to acetylcholine 24 16 3,83 8,58277777777778E-05
GO:0046838 phosphorylated carbohydrate dephosphoryl... 15 10 2,39 0,000820728125
GO:0060732 positive regulation of inositol phosphat... 12 8 1,92 0,000911586956522
GO:0043647 inositol phosphate metabolic process 59 39 9,42 6,50484210526316E-16
GO:0099068 postsynapse assembly 20 13 3,19 2,482875E-05
GO:0045933 positive regulation of muscle contractio... 39 25 6,23 1,42387096774194E-05
GO:0009190 cyclic nucleotide biosynthetic process 63 39 10,06 5,98025806451613E-24
GO:0052652 cyclic purine nucleotide metabolic proce... 63 39 10,06 3,59279069767442E-20
GO:0033622 integrin activation 21 13 3,35 2,57483333333333E-06
GO:0006029 proteoglycan metabolic process 81 50 12,93 5,793375E-18
GO:1903524 positive regulation of blood circulation 57 35 9,1 1,19495027624309E-06
GO:0007631 feeding behavior 85 52 13,57 1,82730107526882E-06
GO:0035150 regulation of tube size 117 70 18,68 1,6406017699115E-09
GO:0050818 regulation of coagulation 74 44 11,81 5,47405511811024E-09
GO:0033762 response to glucagon 32 19 5,11 4,07590638297872E-05
GO:0050795 regulation of behavior 54 32 8,62 7,07587786259542E-09
GO:1900046 regulation of hemostasis 71 42 11,33 1,25747674418605E-11
GO:0030193 regulation of blood coagulation 71 42 11,33 1,23883490566038E-05
GO:0007266 Rho protein signal transduction 165 97 26,34 3,29240983606557E-15
GO:0016486 peptide hormone processing 29 17 4,63 5,72585314685315E-08
GO:0071526 semaphorin-plexin signaling pathway 36 21 5,75 1,17706666666667E-05
GO:0009593 detection of chemical stimulus 424 247 67,68 5,94192307692308E-28
GO:0018126 protein hydroxylation 19 11 3,03 0,0001931125
GO:0007606 sensory perception of chemical stimulus 434 249 69,28 7,44530120481928E-12
GO:0099084 postsynaptic specialization organization 21 12 3,35 2,96836681222707E-05
GO:0010919 regulation of inositol phosphate biosynt... 14 8 2,23 0,000950707692308
GO:0050906 detection of stimulus involved in sensor... 432 246 68,96 9,30901282051282E-13
GO:0007602 phototransduction 53 30 8,46 0,000161407462687
GO:0006874 cellular calcium ion homeostasis 372 210 59,38 1,14996315789474E-08
GO:0006023 aminoglycan biosynthetic process 103 58 16,44 5,94192307692308E-28
GO:0008277 regulation of G-protein coupled receptor... 124 69 19,79 9,655625E-07
GO:0055074 calcium ion homeostasis 379 210 60,5 1,38508275862069E-11
GO:0072376 protein activation cascade 85 47 13,57 2,23152222222222E-11
GO:1901381 positive regulation of potassium ion tra... 31 17 4,95 6,90808943089431E-05
GO:0071709 membrane assembly 24 13 3,83 0,0002151825
GO:0009583 detection of light stimulus 63 34 10,06 2,34565968586387E-06
GO:0051056 regulation of small GTPase mediated sign... 269 145 42,94 5,94192307692308E-28
GO:0043062 extracellular structure organization 351 189 56,03 5,94192307692308E-28
GO:0035987 endodermal cell differentiation 41 22 6,54 6,97454854368932E-06
GO:0001941 postsynaptic membrane organization 28 15 4,47 0,000134085660377
GO:0046839 phospholipid dephosphorylation 34 18 5,43 0,000129231178707
GO:0030574 collagen catabolic process 36 19 5,75 1,78257692307692E-07
4 GO:0009187 cyclic nucleotide metabolic process 78 41 12,45 1,41615833333333E-22
GO:0072503 cellular divalent inorganic cation homeo... 403 210 64,33 1,48768148148148E-08
GO:0051952 regulation of amine transport 75 39 11,97 0,000112463984674
GO:0051606 detection of stimulus 577 300 92,1 5,94192307692308E-28
GO:1904064 positive regulation of cation transmembr... 112 58 17,88 9,60942583732057E-06
GO:1903035 negative regulation of response to wound... 70 36 11,17 1,44303846153846E-06
GO:0008217 regulation of blood pressure 148 76 23,62 2,29440594059406E-10
GO:0019932 second-messenger-mediated signaling 377 193 60,18 5,94192307692308E-28
GO:0030199 collagen fibril organization 43 22 6,86 4,89218333333333E-05
GO:0030239 myofibril assembly 53 27 8,46 5,52831862745098E-06
GO:0072507 divalent inorganic cation homeostasis 417 212 66,56 3,62383950617284E-12
GO:0002027 regulation of heart rate 83 42 13,25 1,76358447488584E-05
GO:0042220 response to cocaine 40 20 6,38 0,000411973333333
GO:0034767 positive regulation of ion transmembrane... 123 61 19,63 1,33891333333333E-07
GO:0015837 amine transport 81 40 12,93 1,01781647058824E-11
GO:0032963 collagen metabolic process 79 39 12,61 1,593178125E-14
GO:0006875 cellular metal ion homeostasis 474 234 75,66 1,02236029411765E-13
GO:0010524 positive regulation of calcium ion trans... 47 23 7,5 2,482875E-05
GO:0003013 circulatory system process 421 206 67,2 0,000375786486486
GO:2000257 regulation of protein activation cascade 45 22 7,18 0,000375786486486
GO:0030048 actin filament-based movement 117 57 18,68 9,655625E-05
GO:0006022 aminoglycan metabolic process 150 73 23,94 5,94192307692308E-28
GO:0006026 aminoglycan catabolic process 60 29 9,58 1,46526597938144E-10
GO:0009581 detection of external stimulus 106 51 16,92 0,000820728125
GO:0043547 positive regulation of GTPase activity 316 151 50,44 7,14308602150538E-11
GO:0030449 regulation of complement activation 44 21 7,02 0,00055535620915
GO:0008360 regulation of cell shape 122 58 19,47 1,5113152173913E-06
GO:0015844 monoamine transport 72 34 11,49 3,21854166666667E-09
GO:0006936 muscle contraction 299 140 47,73 0,000820728125
GO:0043113 receptor clustering 45 21 7,18 6,33155737704918E-05
GO:0033555 multicellular organismal response to str... 60 28 9,58 0,00024848041958
GO:0002687 positive regulation of leukocyte migrati... 99 46 15,8 4,01674E-06
GO:0044091 membrane biogenesis 28 13 4,47 0,000911586956522
GO:0043270 positive regulation of ion transport 212 98 33,84 6,50484210526316E-13
GO:0044057 regulation of system process 422 195 67,36 2,04402153846154E-14
GO:0070588 calcium ion transmembrane transport 246 113 39,27 1,14823648648649E-07
GO:0043087 regulation of GTPase activity 376 171 60,02 3,59771232876712E-13
GO:0055065 metal ion homeostasis 530 241 84,6 1,379375E-09
GO:0007600 sensory perception 781 355 124,66 4,68850248756219E-06
GO:0007215 glutamate receptor signaling pathway 73 33 11,65 1,42387096774194E-05
GO:0030003 cellular cation homeostasis 525 237 83,8 3,6073807106599E-06
GO:0006873 cellular ion homeostasis 538 239 85,88 1,19495027624309E-06
GO:0030168 platelet activation 143 63 22,83 2,3509347826087E-08
GO:0060326 cell chemotaxis 254 111 40,54 6,48858E-17
GO:0051703 intraspecies interaction between organis... 46 20 7,34 0,000144653558052
GO:0060402 calcium ion transport into cytosol 122 53 19,47 8,42672727272727E-10
GO:0051283 negative regulation of sequestering of c... 99 43 15,8 1,76172807017544E-09
GO:0035637 multicellular organismal signaling 153 66 24,42 8,56316E-07
GO:0051282 regulation of sequestering of calcium io... 100 43 15,96 1,90014438502674E-06
GO:0046883 regulation of hormone secretion 225 96 35,91 7,06582170542636E-09
GO:0046847 filopodium assembly 47 20 7,5 4,80347058823529E-05
GO:0051017 actin filament bundle assembly 127 54 20,27 2,39350704225352E-13
GO:0030809 negative regulation of nucleotide biosyn... 33 14 5,27 5,90697058823529E-07
GO:0055082 cellular chemical homeostasis 643 270 102,64 2,14714915254237E-15
GO:0034764 positive regulation of transmembrane tra... 170 71 27,14 9,20591095890411E-08
GO:0070838 divalent metal ion transport 369 154 58,9 5,42554166666667E-07
GO:0002685 regulation of leukocyte migration 144 60 22,99 3,01057435897436E-06
GO:0050817 coagulation 308 128 49,16 1,38689886363636E-11
GO:0032231 regulation of actin filament bundle asse... 82 34 13,09 0,000284304513889
GO:0072511 divalent inorganic cation transport 372 154 59,38 1,17476770833333E-10
GO:0051705 multi-organism behavior 58 24 9,26 0,000693224358974
GO:0007229 integrin-mediated signaling pathway 92 38 14,69 2,79034801762115E-05
GO:0007492 endoderm development 68 28 10,85 0,000647861290323
GO:0002920 regulation of humoral immune response 61 25 9,74 5,42630165289256E-05
GO:0046887 positive regulation of hormone secretion 110 45 17,56 9,655625E-05
GO:0060359 response to ammonium ion 103 42 16,44 8,00147410358566E-05
GO:0019751 polyol metabolic process 108 44 17,24 1,19495027624309E-06
GO:1903317 regulation of protein maturation 95 38 15,16 4,33325609756098E-12
GO:0042698 ovulation cycle 55 22 8,78 0,000950707692308
GO:0070613 regulation of protein processing 93 37 14,84 0,00024848041958
GO:0003008 system process 1720 684 274,55 5,94192307692308E-28
GO:0010522 regulation of calcium ion transport into... 83 33 13,25 0,000340943448276
4 GO:0006956 complement activation 63 25 10,06 0,000201508695652
GO:0050801 ion homeostasis 650 257 103,75 0,000992137614679
GO:0007264 small GTPase mediated signal transductio... 466 184 74,38 1,41171896551724E-15
GO:1904062 regulation of cation transmembrane trans... 257 101 41,02 0,000733338607595
GO:0099173 postsynapse organization 107 42 17,08 1,59473548387097E-07
GO:0002090 regulation of receptor internalization 36 14 5,75 0,000693224358974
GO:0010927 cellular component assembly involved in ... 85 33 13,57 4,84801255230126E-05
GO:1903034 regulation of response to wounding 129 50 20,59 8,17014423076923E-06
GO:0009914 hormone transport 265 102 42,3 5,76762666666667E-13
GO:0050878 regulation of body fluid levels 438 168 69,91 8,42672727272727E-10
GO:0050900 leukocyte migration 361 138 57,62 4,93053191489362E-18
GO:0034765 regulation of ion transmembrane transpor... 379 144 60,5 2,48755084745763E-09
GO:0032964 collagen biosynthetic process 32 12 5,11 0,000992137614679
GO:0030029 actin filament-based process 619 230 98,8 5,94192307692308E-28
GO:0099536 synaptic signaling 547 203 87,31 8,07964942528736E-07
GO:0043269 regulation of ion transport 537 199 85,72 3,14701851851852E-16
GO:0010559 regulation of glycoprotein biosynthetic ... 30 11 4,79 0,000182423616236
GO:0046683 response to organophosphorus 118 43 18,84 9,655625E-05
GO:0071804 cellular potassium ion transport 179 65 28,57 1,11185984848485E-08
GO:0030036 actin cytoskeleton organization 541 196 86,35 3,15285714285714E-17
GO:0019725 cellular homeostasis 784 284 125,14 5,94192307692308E-28
GO:0014074 response to purine-containing compound 133 48 21,23 1,42387096774194E-05
GO:0048878 chemical homeostasis 932 329 148,77 5,94192307692308E-28
GO:0099177 regulation of trans-synaptic signaling 301 106 48,05 0,000601909090909
GO:0032970 regulation of actin filament-based proce... 316 111 50,44 1,37788378378378E-09
GO:1900371 regulation of purine nucleotide biosynth... 83 29 13,25 8,00147410358566E-05
GO:0034762 regulation of transmembrane transport 458 159 73,11 1,48184285714286E-10
GO:0034330 cell junction organization 249 86 39,75 3,62383950617284E-12
GO:0030808 regulation of nucleotide biosynthetic pr... 84 29 13,41 9,69671276595745E-11
GO:0007610 behavior 461 159 73,58 8,09233333333333E-21
GO:0050435 amyloid-beta metabolic process 35 12 5,59 3,76321794871795E-05
GO:0042330 taxis 540 185 86,19 3,11576470588235E-09
GO:0090066 regulation of anatomical structure size 415 142 66,24 0,000123827862595
GO:0006935 chemotaxis 538 184 85,88 2,01508695652174E-09
GO:0045980 negative regulation of nucleotide metabo... 41 14 6,54 8,00147410358566E-05
GO:0051345 positive regulation of hydrolase activit... 618 208 98,65 2,80013125E-07
GO:0032103 positive regulation of response to exter... 242 81 38,63 1,37164018691589E-05
GO:0016485 protein processing 217 72 34,64 9,74758333333333E-12
GO:1903531 negative regulation of secretion by cell 160 53 25,54 1,76358447488584E-05
GO:0043112 receptor metabolic process 152 50 24,26 2,93335443037975E-12
GO:0034329 cell junction assembly 207 68 33,04 2,42337254901961E-10
GO:0009611 response to wounding 562 184 89,71 1,01217586206897E-25
GO:0031345 negative regulation of cell projection o... 134 43 21,39 0,000950707692308
GO:0030001 metal ion transport 711 228 113,49 0,000112463984674
GO:0010466 negative regulation of peptidase activit... 191 61 30,49 9,15948616600791E-05
GO:0097435 supramolecular fiber organization 537 171 85,72 1,5449E-21
GO:0051271 negative regulation of cellular componen... 253 80 40,38 3,90126262626263E-06
GO:0010951 negative regulation of endopeptidase act... 187 59 29,85 1,04150561797753E-06
GO:1903530 regulation of secretion by cell 578 181 92,26 1,70112584269663E-11
GO:0010469 regulation of signaling receptor activit... 477 149 76,14 7,46328502415459E-06
GO:0019730 antimicrobial humoral response 103 32 16,44 0,000647861290323
GO:0022604 regulation of cell morphogenesis 375 116 59,86 2,7205786163522E-07
GO:0034440 lipid oxidation 94 29 15 0,000139389473684
GO:0040013 negative regulation of locomotion 261 80 41,66 4,89473267326733E-06
GO:0051270 regulation of cellular component movemen... 787 240 125,62 1,16971E-13
GO:0051336 regulation of hydrolase activity 1017 309 162,33 4,66384905660377E-10
GO:0040012 regulation of locomotion 786 238 125,46 1,09710289855072E-13
GO:0098655 cation transmembrane transport 687 208 109,66 2,79034801762115E-05
GO:0050808 synapse organization 261 79 41,66 5,14966666666667E-08
GO:0015850 organic hydroxy compound transport 202 61 32,24 3,05165432098765E-07
GO:0007163 establishment or maintenance of cell pol... 176 53 28,09 0,00024848041958
GO:1903532 positive regulation of secretion by cell 320 96 51,08 2,64001898734177E-07
GO:0006954 inflammatory response 591 177 94,34 2,08327424242424E-14
GO:0051668 localization within membrane 107 32 17,08 0,000284304513889
GO:0002790 peptide secretion 478 142 76,3 2,4451654676259E-08
GO:0046718 viral entry into host cell 108 32 17,24 0,000134085660377
GO:0040017 positive regulation of locomotion 446 132 71,19 4,32572E-09
GO:1902903 regulation of supramolecular fiber organ... 289 85 46,13 0,000359771232877
GO:0022610 biological adhesion 1120 329 178,77 5,94192307692308E-28
GO:0051272 positive regulation of cellular componen... 429 126 68,48 1,48768148148148E-08
GO:1900542 regulation of purine nucleotide metaboli... 99 29 15,8 0,000224613120567
GO:0032101 regulation of response to external stimu... 630 183 100,56 2,30686425339367E-05
4 GO:0032940 secretion by cell 1227 353 195,85 2,05986666666667E-27
GO:0051674 localization of cell 1280 368 204,31 4,56447727272727E-20
GO:0051604 protein maturation 272 78 43,42 1,17706666666667E-14
GO:0006812 cation transport 918 261 146,53 1,96623636363636E-05
GO:0051050 positive regulation of transport 808 229 128,97 3,0898E-19
GO:0044272 sulfur compound biosynthetic process 166 47 26,5 4,58376923076923E-11
GO:0006140 regulation of nucleotide metabolic proce... 103 29 16,44 3,54696428571429E-06
GO:0046903 secretion 1342 377 214,21 1,99928235294118E-23
GO:0032989 cellular component morphogenesis 890 247 142,06 1,91942121212121E-23
GO:0040011 locomotion 1472 407 234,96 5,94192307692308E-28
GO:0051051 negative regulation of transport 384 106 61,29 5,72585314685315E-08
GO:0016054 organic acid catabolic process 247 68 39,43 3,67833333333333E-10
GO:0046395 carboxylic acid catabolic process 247 68 39,43 7,47532258064516E-05
GO:0009306 protein secretion 451 123 71,99 6,90808943089431E-05
GO:0006928 movement of cell or subcellular componen... 1669 454 266,41 5,94192307692308E-28
GO:0045861 negative regulation of proteolysis 276 75 44,06 2,2447264957265E-09
GO:0051049 regulation of transport 1476 398 235,6 3,14701851851852E-16
GO:1990778 protein localization to cell periphery 230 62 36,71 2,99980582524272E-10
GO:0032879 regulation of localization 2182 583 348,29 5,94192307692308E-28
GO:0031960 response to corticosteroid 131 35 20,91 0,0002151825
GO:0023052 signaling 5319 1409 849,02 5,94192307692308E-28
GO:0007165 signal transduction 4969 1312 793,15 5,94192307692308E-28
GO:0007154 cell communication 5349 1397 853,81 5,94192307692308E-28
GO:0051241 negative regulation of multicellular org... 919 234 146,69 0,000497858139535
GO:0006809 nitric oxide biosynthetic process 55 14 8,78 0,000374361433447
GO:0001568 blood vessel development 560 142 89,39 0,000375786486486
GO:0098657 import into cell 650 164 103,75 0,000211173381295
GO:0001944 vasculature development 585 145 93,38 0,00055535620915
GO:0065008 regulation of biological quality 3134 776 500,25 1,83456875E-23
GO:0051261 protein depolymerization 85 21 13,57 1,5113152173913E-06
GO:0072358 cardiovascular system development 593 146 94,65 0,000733338607595
GO:0048646 anatomical structure formation involved ... 890 218 142,06 4,01674E-06
GO:0023051 regulation of signaling 2907 703 464,02 4,4851935483871E-15
GO:0051493 regulation of cytoskeleton organization 440 106 70,23 0,000112463984674
GO:0010646 regulation of cell communication 2879 690 459,55 7,37862686567164E-14
GO:0009653 anatomical structure morphogenesis 2056 488 328,18 2,31735E-16
GO:0051239 regulation of multicellular organismal p... 2354 549 375,75 4,32572E-09
GO:0051716 cellular response to stimulus 6086 1408 971,45 5,94192307692308E-28
GO:0032501 multicellular organismal process 5860 1350 935,37 5,94192307692308E-28
GO:0051223 regulation of protein transport 560 129 89,39 6,38110869565217E-11
GO:0006790 sulfur compound metabolic process 323 74 51,56 7,99086206896552E-08
GO:0065009 regulation of molecular function 2633 603 420,28 6,27615625E-09
GO:0018958 phenol-containing compound metabolic pro... 83 19 13,25 0,000219914590747
GO:0009101 glycoprotein biosynthetic process 282 64 45,01 2,78940277777778E-13
GO:0044242 cellular lipid catabolic process 180 40 28,73 0,000820728125
GO:0046488 phosphatidylinositol metabolic process 205 45 32,72 0,000345080756014
GO:0009100 glycoprotein metabolic process 342 75 54,59 1,86962420382166E-07
GO:0044282 small molecule catabolic process 394 86 62,89 4,49781012658228E-05
GO:0070201 regulation of establishment of protein l... 601 131 95,93 3,31961983471074E-09
GO:0048583 regulation of response to stimulus 3365 729 537,12 2,13089655172414E-09
GO:0050896 response to stimulus 7323 1575 1168,9 5,94192307692308E-28
GO:0051258 protein polymerization 229 49 36,55 2,27191176470588E-16
GO:1901617 organic hydroxy compound biosynthetic pr... 225 48 35,91 1,95145263157895E-06
GO:1901615 organic hydroxy compound metabolic proce... 445 94 71,03 1,51460784313725E-07
GO:0062012 regulation of small molecule metabolic p... 322 67 51,4 0,000201508695652
GO:0001934 positive regulation of protein phosphory... 849 174 135,52 0,00039918557047
GO:0010562 positive regulation of phosphorus metabo... 954 195 152,28 1,02451263157895E-10
GO:0045937 positive regulation of phosphate metabol... 954 195 152,28 0,000336777508651
GO:0006952 defense response 1288 257 205,59 2,91362719298246E-05
GO:1901137 carbohydrate derivative biosynthetic pro... 628 125 100,24 4,21336363636364E-16
GO:0006164 purine nucleotide biosynthetic process 215 42 34,32 0,00055535620915
GO:0019220 regulation of phosphate metabolic proces... 1448 281 231,13 2,79034801762115E-05
GO:0051174 regulation of phosphorus metabolic proce... 1456 282 232,41 2,21296486486486E-22
GO:0051179 localization 5214 997 832,26 3,75786486486487E-13
4 GO:0001932 regulation of protein phosphorylation 1195 228 190,75 3,01057435897436E-06
GO:0050789 regulation of biological process 9094 1727 1451,59 5,94192307692308E-28
GO:0050794 regulation of cellular process 8535 1618 1362,36 8,75443333333333E-25
GO:0065007 biological regulation 9603 1790 1532,83 5,94192307692308E-28
GO:0044255 cellular lipid metabolic process 927 171 147,97 3,67833333333333E-10
GO:0072522 purine-containing compound biosynthetic ... 228 42 36,39 3,7130643776824E-05
GO:0006082 organic acid metabolic process 968 177 154,51 3,41904098360656E-09
GO:0032502 developmental process 4760 860 759,79 3,66247844827586E-05
GO:0031401 positive regulation of protein modificat... 1032 185 164,73 1,59473548387097E-07
GO:0009617 response to bacterium 504 90 80,45 0,000733338607595
GO:0044281 small molecule metabolic process 1779 315 283,96 5,241625E-16
GO:0006468 protein phosphorylation 1629 287 260,02 2,83231666666667E-15
GO:0072657 protein localization to membrane 483 85 77,1 0,000112463984674
GO:0016053 organic acid biosynthetic process 366 64 58,42 0,000211173381295
GO:0044283 small molecule biosynthetic process 659 109 105,19 6,63176585365854E-06
GO:0009987 cellular process 12542 2071 2001,96 8,42672727272727E-10
GO:0006793 phosphorus metabolic process 2768 457 441,83 5,2968E-23
GO:0031399 regulation of protein modification proce... 1515 248 241,82 7,09576162790698E-07
GO:0090407 organophosphate biosynthetic process 569 92 90,82 1,95145263157895E-06
GO:0032270 positive regulation of cellular protein ... 1338 215 213,57 2,64153886010363E-06
GO:0051246 regulation of protein metabolic process 2330 349 371,92 2,87869565217391E-07
GO:0032268 regulation of cellular protein metabolic... 2170 322 346,38 1,87260606060606E-10
GO:1901293 nucleoside phosphate biosynthetic proces... 284 42 45,33 0,000166550557621
GO:0009259 ribonucleotide metabolic process 397 58 63,37 6,33155737704918E-05
GO:1901564 organonitrogen compound metabolic proces... 5705 796 910,63 6,02887804878049E-21
GO:0019538 protein metabolic process 4819 653 769,21 5,77532710280374E-10
GO:0044267 cellular protein metabolic process 4335 525 691,95 0,000733338607595
GO:0051666 actin cortical patch localization 5 5 0,17 2,32899497487437E-05
GO:0000147 actin cortical patch assembly 5 5 0,17 2,47184E-05
GO:0044396 actin cortical patch organization 5 5 0,17 2,75328712871287E-05
GO:0098943 neurotransmitter receptor transport, pos... 6 5 0,2 5,44819806763285E-05
GO:0098953 receptor diffusion trapping 6 5 0,2 0,000549879661017
GO:1905247 positive regulation of aspartic-type pep... 6 5 0,2 0,000886418032787
B GO:0038128 ERBB2 signaling pathway 28 22 0,93 1,7745472972973E-07
GO:0048013 ephrin receptor signaling pathway 76 53 2,54 1,158675E-10
GO:0036092 phosphatidylinositol-3-phosphate biosynt... 49 33 1,64 2,13089655172414E-11
GO:0002433 immune response-regulating cell surface ... 67 45 2,24 2,00837E-26
GO:0002431 Fc receptor mediated stimulatory signali... 72 47 2,4 6,70213970588235E-09
GO:0046854 phosphatidylinositol phosphorylation 97 57 3,24 5,47152083333333E-14
GO:0038093 Fc receptor signaling pathway 112 62 3,74 6,8779794520548E-08
GO:0045730 respiratory burst 24 13 0,8 1,28965565217391E-11
GO:0046834 lipid phosphorylation 109 57 3,64 5,5175E-26
GO:0061098 positive regulation of protein tyrosine ... 40 19 1,34 0,000357553275109
GO:0051897 positive regulation of protein kinase B ... 136 61 4,54 2,23603947368421E-17
GO:0038179 neurotrophin signaling pathway 29 13 0,97 0,000549879661017
GO:0031295 T cell costimulation 49 21 1,64 0,000214761883408
GO:0014068 positive regulation of phosphatidylinosi... 66 28 2,2 3,35464E-06
GO:0031294 lymphocyte costimulation 50 21 1,67 1,55601438848921E-08
GO:0018108 peptidyl-tyrosine phosphorylation 333 122 11,11 4,68151515151515E-28
GO:0018212 peptidyl-tyrosine modification 335 122 11,18 4,68151515151515E-28
GO:0051896 regulation of protein kinase B signaling 182 66 6,07 1,80634461538462E-19
GO:0061097 regulation of protein tyrosine kinase ac... 67 24 2,24 0,000246090265487
GO:0002768 immune response-regulating cell surface ... 269 92 8,98 1,15165272727273E-22
GO:0002429 immune response-activating cell surface ... 244 83 8,14 1,13734969325153E-06
GO:0043491 protein kinase B signaling 203 69 6,78 1,2793703125E-19
GO:0006801 superoxide metabolic process 56 19 1,87 0,000541375213675
GO:0046488 phosphatidylinositol metabolic process 205 67 6,84 1,16221834862385E-12
GO:0007169 transmembrane receptor protein tyrosine ... 598 194 19,96 2,06958301886792E-23
GO:0043303 mast cell degranulation 39 12 1,3 1,38959259259259E-05
GO:0070374 positive regulation of ERK1 and ERK2 cas... 196 60 6,54 2,69012437810945E-05
GO:0030032 lamellipodium assembly 56 17 1,87 1,98064102564103E-05
GO:0048017 inositol lipid-mediated signaling 145 44 4,84 7,17709448818898E-10
GO:0014066 regulation of phosphatidylinositol 3-kin... 96 29 3,2 1,61769633507853E-05
GO:0097581 lamellipodium organization 70 21 2,34 0,000264259210526
GO:0006909 phagocytosis 207 62 6,91 9,62763768115942E-09
GO:0060193 positive regulation of lipase activity 63 18 2,1 0,0002207
GO:0050730 regulation of peptidyl-tyrosine phosphor... 202 57 6,74 9,53641975308642E-07
GO:0046777 protein autophosphorylation 212 59 7,08 4,56942253521127E-08
GO:0032273 positive regulation of protein polymeriz... 113 31 3,77 5,20752808988764E-06
GO:0070372 regulation of ERK1 and ERK2 cascade 258 67 8,61 1,09150543478261E-05
GO:0051781 positive regulation of cell division 78 20 2,6 4,74003409090909E-06
GO:0044344 cellular response to fibroblast growth f... 117 30 3,91 1,09150543478261E-05
GO:0097485 neuron projection guidance 223 57 7,44 1,05095238095238E-07
GO:0002764 immune response-regulating signaling pat... 399 98 13,32 4,99820588235294E-28
GO:0030258 lipid modification 262 64 8,74 6,33155737704918E-21
B GO:0071774 response to fibroblast growth factor 123 30 4,11 1,8410725388601E-05
GO:0010634 positive regulation of epithelial cell m... 121 29 4,04 7,24858293838863E-05
GO:0007167 enzyme linked receptor protein signaling... 869 203 29,01 4,68151515151515E-28
GO:0010324 membrane invagination 56 13 1,87 0,000651856540084
GO:0032956 regulation of actin cytoskeleton organiz... 285 63 9,51 9,38889502762431E-06
GO:0060996 dendritic spine development 68 15 2,27 0,000829904958678
GO:0051495 positive regulation of cytoskeleton orga... 198 43 6,61 3,10986363636364E-07
GO:0006650 glycerophospholipid metabolic process 311 67 10,38 8,01599056603774E-05
GO:0032418 lysosome localization 56 12 1,87 0,000116048826291
GO:0043410 positive regulation of MAPK cascade 461 97 15,39 3,10755747126437E-06
GO:0032271 regulation of protein polymerization 177 37 5,91 1,23922459893048E-05
GO:1902905 positive regulation of supramolecular fi... 179 37 5,97 0,000382866521739
GO:0099173 postsynapse organization 107 22 3,57 1,40445454545455E-06
GO:0006935 chemotaxis 538 110 17,96 4,68151515151515E-28
GO:0042330 taxis 540 110 18,02 4,91559090909091E-19
GO:0002253 activation of immune response 446 89 14,89 3,33557954545455E-15
GO:0032970 regulation of actin filament-based proce... 316 63 10,55 2,26585333333333E-21
GO:1900046 regulation of hemostasis 71 14 2,37 0,000147467727273
GO:0010632 regulation of epithelial cell migration 185 36 6,17 1,22461585365854E-06
GO:0090130 tissue migration 240 46 8,01 4,8554E-13
GO:0006644 phospholipid metabolic process 398 75 13,28 2,59761946902655E-12
GO:0000165 MAPK cascade 773 144 25,8 3,57964634146341E-16
GO:0033674 positive regulation of kinase activity 481 88 16,05 0,000233451555556
GO:0023014 signal transduction by protein phosphory... 789 144 26,34 4,68151515151515E-28
GO:0061564 axon development 409 74 13,65 0,000446156465517
GO:0050867 positive regulation of cell activation 274 49 9,15 4,87863157894737E-09
GO:0032535 regulation of cellular component size 305 54 10,18 2,40317777777778E-14
GO:0030036 actin cytoskeleton organization 541 95 18,06 9,30092857142857E-14
GO:0051302 regulation of cell division 143 25 4,77 6,19928025477707E-07
GO:0051258 protein polymerization 229 40 7,64 1,23592E-13
GO:0046486 glycerolipid metabolic process 384 67 12,82 2,72629411764706E-13
GO:0040017 positive regulation of locomotion 446 77 14,89 3,86225E-14
GO:1902903 regulation of supramolecular fiber organ... 289 49 9,65 2,62000584795322E-06
GO:0043408 regulation of MAPK cascade 632 106 21,09 8,44417391304348E-07
GO:1903409 reactive oxygen species biosynthetic pro... 85 14 2,84 1,931125E-06
GO:0048667 cell morphogenesis involved in neuron di... 474 78 15,82 0,000147467727273
GO:0045017 glycerolipid biosynthetic process 232 38 7,74 5,99513432835821E-19
GO:0006897 endocytosis 576 94 19,23 5,24683018867924E-13
GO:0051347 positive regulation of transferase activ... 544 88 18,16 1,48906024096386E-06
GO:0030029 actin filament-based process 619 100 20,66 4,68151515151515E-28
GO:0001934 positive regulation of protein phosphory... 849 137 28,34 1,16282795698925E-05
GO:1902533 positive regulation of intracellular sig... 892 142 29,77 7,0698813559322E-11
GO:0048858 cell projection morphogenesis 535 85 17,86 1,09150543478261E-05
GO:0045785 positive regulation of cell adhesion 341 54 11,38 2,1913475177305E-08
GO:0008654 phospholipid biosynthetic process 240 38 8,01 1,23592E-17
GO:0042327 positive regulation of phosphorylation 891 141 29,74 1,931125E-06
GO:0050778 positive regulation of immune response 589 93 19,66 4,67238048780488E-05
GO:0050776 regulation of immune response 756 119 25,23 4,93053191489362E-14
GO:0031334 positive regulation of protein complex a... 225 35 7,51 2,28582142857143E-05
GO:0000904 cell morphogenesis involved in different... 603 93 20,13 3,11471774193548E-10
GO:0050865 regulation of cell activation 436 67 14,55 1,01689620253165E-16
GO:0045937 positive regulation of phosphate metabol... 954 146 31,84 1,06114343434343E-13
GO:0090066 regulation of anatomical structure size 415 63 13,85 1,11124385964912E-11
GO:0051493 regulation of cytoskeleton organization 440 66 14,69 5,43378620689655E-08
GO:2000377 regulation of reactive oxygen species me... 141 21 4,71 4,87468446601942E-05
GO:0001525 angiogenesis 424 63 14,15 7,9176125E-07
GO:0098657 import into cell 650 96 21,7 1,02993333333333E-22
GO:0006809 nitric oxide biosynthetic process 55 8 1,84 0,000886418032787
GO:0072593 reactive oxygen species metabolic proces... 222 32 7,41 4,078536E-10
GO:0070848 response to growth factor 603 86 20,13 9,08764705882353E-11
GO:0071363 cellular response to growth factor stimu... 576 82 19,23 2,96702649006623E-07
GO:0007163 establishment or maintenance of cell pol... 176 25 5,87 1,9700067114094E-07
GO:0030155 regulation of cell adhesion 552 78 18,42 6,58854411764706E-19
GO:0002684 positive regulation of immune system pro... 814 115 27,17 3,41081818181818E-17
GO:0040012 regulation of locomotion 786 110 26,23 4,47207894736842E-27
GO:0018193 peptidyl-amino acid modification 1026 141 34,25 1,9863E-22
GO:0051270 regulation of cellular component movemen... 787 108 26,27 9,97747916666667E-25
GO:0040011 locomotion 1472 200 49,13 4,68151515151515E-28
GO:0032989 cellular component morphogenesis 890 119 29,71 1,85388E-25
B GO:0051674 localization of cell 1280 170 42,72 4,68151515151515E-28
GO:0001568 blood vessel development 560 74 18,69 5,64926119402985E-09
GO:0010469 regulation of signaling receptor activit... 477 63 15,92 4,61955392156863E-05
GO:0031175 neuron projection development 762 100 25,43 0,000141733944954
GO:0071375 cellular response to peptide hormone sti... 283 37 9,45 0,000769244813278
GO:0001944 vasculature development 585 76 19,53 2,26585333333333E-07
GO:0009967 positive regulation of signal transducti... 1337 173 44,63 4,93509722222222E-18
GO:0008037 cell recognition 116 15 3,87 0,000708079166667
GO:0051338 regulation of transferase activity 803 103 26,8 6,1796E-05
GO:0072358 cardiovascular system development 593 76 19,79 4,18618064516129E-07
GO:0050817 coagulation 308 39 10,28 1,3148085106383E-05
GO:0048584 positive regulation of response to stimu... 1878 231 62,68 4,68151515151515E-28
GO:0050808 synapse organization 261 32 8,71 0,000214761883408
GO:0008284 positive regulation of cell proliferatio... 751 92 25,07 2,83035114503817E-09
GO:0010647 positive regulation of cell communicatio... 1464 179 48,87 8,07561363636364E-26
GO:0023056 positive regulation of signaling 1470 179 49,07 4,47207894736842E-27
GO:0007166 cell surface receptor signaling pathway 2446 297 81,64 4,68151515151515E-28
GO:0097435 supramolecular fiber organization 537 65 17,92 1,43711627906977E-09
GO:0002682 regulation of immune system process 1169 140 39,02 4,68151515151515E-28
GO:0043066 negative regulation of apoptotic process 754 90 25,17 5,98741148325359E-05
GO:0043254 regulation of protein complex assembly 386 46 12,88 0,000167771945701
GO:0043069 negative regulation of programmed cell d... 763 90 25,47 2,67901734104046E-06
GO:0048666 neuron development 863 101 28,81 1,98064102564103E-05
GO:0009611 response to wounding 562 65 18,76 5,72185185185185E-09
GO:0016310 phosphorylation 1979 226 66,05 2,0690625E-12
GO:1902531 regulation of intracellular signal trans... 1562 178 52,14 2,36278823529412E-06
GO:0043085 positive regulation of catalytic activit... 1159 130 38,68 1,5449E-05
GO:0010638 positive regulation of organelle organiz... 537 60 17,92 2,09665E-08
GO:0060548 negative regulation of cell death 828 91 27,64 3,44977669902913E-13
GO:0072359 circulatory system development 860 93 28,7 2,32899497487437E-05
GO:0060627 regulation of vesicle-mediated transport 391 42 13,05 1,16282795698925E-05
GO:0030030 cell projection organization 1197 128 39,95 1,21086756756757E-17
GO:0051247 positive regulation of protein metabolic... 1428 151 47,66 4,68151515151515E-28
GO:0035556 intracellular signal transduction 2361 247 78,81 5,47152083333333E-14
GO:0035295 tube development 833 86 27,8 2,32899497487437E-05
GO:0001775 cell activation 1152 118 38,45 6,46702325581395E-26
GO:0051094 positive regulation of developmental pro... 1072 108 35,78 6,49838888888889E-10
GO:0030182 neuron differentiation 1051 105 35,08 0,000708079166667
GO:0044087 regulation of cellular component biogene... 776 76 25,9 1,64620491803279E-10
GO:0006955 immune response 1656 162 55,27 1,1035E-15
GO:0022008 neurogenesis 1241 121 41,42 5,94192307692308E-05
GO:0051240 positive regulation of multicellular org... 1315 125 43,89 9,87019444444444E-13
GO:0009719 response to endogenous stimulus 1405 131 46,9 1,43711627906977E-09
GO:0009653 anatomical structure morphogenesis 2056 191 68,62 3,86225E-13
GO:0042127 regulation of cell proliferation 1301 119 43,42 1,68534545454545E-12
GO:0023057 negative regulation of signaling 1106 101 36,92 2,67901734104046E-06
GO:0010648 negative regulation of cell communicatio... 1102 100 36,78 1,8410725388601E-05
GO:0019637 organophosphate metabolic process 965 86 32,21 2,6701975308642E-16
GO:0009966 regulation of signal transduction 2612 232 87,18 4,93509722222222E-08
GO:0016192 vesicle-mediated transport 1644 145 54,87 8,85279775280899E-15
GO:0044255 cellular lipid metabolic process 927 81 30,94 3,98683870967742E-14
GO:0010941 regulation of cell death 1376 120 45,93 2,37676923076923E-11
GO:0032940 secretion by cell 1227 107 40,95 9,52335616438356E-18
GO:0009605 response to external stimulus 1929 168 64,39 1,1746347107438E-10
GO:0002376 immune system process 2380 207 79,44 4,68151515151515E-28
GO:0048468 cell development 1653 143 55,17 2,8158275862069E-05
GO:0048585 negative regulation of response to stimu... 1294 109 43,19 4,86157342657343E-08
GO:0051050 positive regulation of transport 808 68 26,97 0,000708079166667
GO:0008283 cell proliferation 1662 139 55,47 6,83321153846154E-24
B GO:0046903 secretion 1342 110 44,79 5,54579487179487E-07
GO:0071310 cellular response to organic substance 2238 182 74,7 7,20953333333333E-06
GO:0070838 divalent metal ion transport 369 30 12,32 2,2853550295858E-06
GO:0051128 regulation of cellular component organiz... 2018 164 67,36 2,09665E-18
GO:0072511 divalent inorganic cation transport 372 30 12,42 0,000945857142857
GO:0033043 regulation of organelle organization 1065 85 35,55 0,000128148387097
GO:0008219 cell death 1804 141 60,21 5,14966666666667E-20
GO:0090407 organophosphate biosynthetic process 569 44 18,99 4,68151515151515E-09
GO:0045595 regulation of cell differentiation 1377 105 45,96 6,45337974683544E-07
GO:0051301 cell division 538 41 17,96 0,000128148387097
GO:0050793 regulation of developmental process 1985 151 66,26 5,77532710280374E-13
GO:0070887 cellular response to chemical stimulus 2671 199 89,15 0,000122154883721
GO:0048869 cellular developmental process 3276 232 109,35 4,68151515151515E-28
GO:0023052 signaling 5319 372 177,54 4,68151515151515E-28
GO:0048522 positive regulation of cellular process 4294 288 143,32 4,68151515151515E-28
GO:0036211 protein modification process 3414 228 113,95 5,14966666666667E-17
GO:0006464 cellular protein modification process 3414 228 113,95 2,38643089430894E-10
GO:0048518 positive regulation of biological proces... 4802 316 160,28 4,68151515151515E-28
GO:0040007 growth 764 50 25,5 0,000251811894273
GO:0031325 positive regulation of cellular metaboli... 2736 176 91,32 1,95332183908046E-15
GO:0043412 macromolecule modification 3579 228 119,46 4,68151515151515E-28
GO:0007275 multicellular organism development 4107 258 137,08 5,52366480446927E-06
GO:0009893 positive regulation of metabolic process 2921 181 97,5 3,56515384615385E-14
GO:0051173 positive regulation of nitrogen compound... 2648 163 88,38 2,82869014084507E-18
GO:0010604 positive regulation of macromolecule met... 2751 166 91,82 2,60032673267327E-13
GO:0044267 cellular protein metabolic process 4335 241 144,69 6,24534042553191E-25
GO:0016043 cellular component organization 5129 265 171,19 1,47304418604651E-15
GO:0071840 cellular component organization or bioge... 5277 265 176,13 1,12686823529412E-15
B GO:0048523 negative regulation of cellular process 3779 188 126,13 3,10986363636364E-07
GO:0048519 negative regulation of biological proces... 4189 198 139,82 3,10986363636364E-07
GO:0080090 regulation of primary metabolic process 4805 200 160,38 0,000428024242424
GO:0044260 cellular macromolecule metabolic process 6674 260 222,76 0,000122154883721
GO:0043170 macromolecule metabolic process 7510 275 250,67 0,000503915879828
GO:0048490 anterograde synaptic vesicle transport 18 18 1,33 2,30379824561404E-08
GO:0099517 synaptic vesicle transport along microtu... 18 18 1,33 4,414E-07
GO:0099514 synaptic vesicle cytoskeletal transport 18 18 1,33 1,30292771084337E-05
GO:0035333 Notch receptor processing, ligand-depend... 7 7 0,52 6,84852577319588E-05
GO:0044340 canonical Wnt signaling pathway involved... 6 6 0,44 0,000275119178082
GO:0031629 synaptic vesicle fusion to presynaptic a... 15 14 1,11 6,517546875E-05
GO:0016082 synaptic vesicle priming 13 11 0,96 3,30207633587786E-07
GO:0048268 clathrin coat assembly 16 13 1,18 6,35125555555556E-10
GO:0007220 Notch receptor processing 10 8 0,74 1,30292771084337E-05
GO:0090110 cargo loading into COPII-coated vesicle 14 11 1,03 0,000161846666667
GO:1904948 midbrain dopaminergic neuron differentia... 16 12 1,18 3,91374666666667E-06
GO:0090177 establishment of planar polarity involve... 12 9 0,88 7,76331658291457E-05
GO:0035459 cargo loading into vesicle 19 14 1,4 5,95166393442623E-08
GO:0006896 Golgi to vacuole transport 15 11 1,11 0,000468751709402
GO:0048278 vesicle docking 58 40 4,28 1,90840588235294E-12
GO:0072583 clathrin-dependent endocytosis 34 23 2,51 5,49014049586777E-08
GO:0035493 SNARE complex assembly 15 10 1,11 2,85470652173913E-05
GO:0048208 COPII vesicle coating 63 40 4,64 3,23024545454545E-22
GO:0042228 interleukin-8 biosynthetic process 13 8 0,96 5,793375E-06
GO:0140029 exocytic process 66 39 4,87 2,16286E-24
GO:0032802 low-density lipoprotein particle recepto... 12 7 0,88 9,89344827586207E-05
GO:0006900 vesicle budding from membrane 93 54 6,86 1,47773043478261E-13
GO:0006906 vesicle fusion 72 41 5,31 1,7931875E-08
GO:0016339 calcium-dependent cell-cell adhesion via... 25 14 1,84 2,00749717514124E-05
GO:0048489 synaptic vesicle transport 99 54 7,3 5,25266E-13
GO:0006903 vesicle targeting 85 46 6,27 3,2829125E-26
GO:0090174 organelle membrane fusion 76 41 5,6 3,54909459459459E-06
5 GO:0036465 synaptic vesicle recycling 51 27 3,76 9,36801063829787E-10
GO:0099003 vesicle-mediated transport in synapse 126 66 9,29 1,02993333333333E-11
GO:0090175 regulation of establishment of planar po... 56 29 4,13 2,15768156424581E-05
GO:0044331 cell-cell adhesion mediated by cadherin 34 17 2,51 5,80359259259259E-05
GO:0048488 synaptic vesicle endocytosis 40 20 2,95 0,000202136448598
GO:0071542 dopaminergic neuron differentiation 30 15 2,21 0,000756685714286
GO:0086010 membrane depolarization during action po... 35 17 2,58 0,000174073239437
GO:0006888 ER to Golgi vesicle-mediated transport 188 90 13,86 1,09430416666667E-18
GO:0099504 synaptic vesicle cycle 124 59 9,14 5,10488695652174E-19
GO:0051767 nitric-oxide synthase biosynthetic proce... 17 8 1,25 0,000394584848485
GO:0051650 establishment of vesicle localization 221 100 16,29 1,25523125E-20
GO:0042094 interleukin-2 biosynthetic process 20 9 1,47 2,00749717514124E-05
GO:0051648 vesicle localization 232 104 17,1 4,55338947368421E-20
GO:0007164 establishment of tissue polarity 67 30 4,94 2,79198795180723E-10
GO:0045454 cell redox homeostasis 63 28 4,64 5,42070175438596E-12
GO:0050690 regulation of defense response to virus ... 27 12 1,99 8,49695E-05
GO:0016050 vesicle organization 278 122 20,5 2,207E-27
GO:0048284 organelle fusion 94 41 6,93 4,8554E-16
GO:0032482 Rab protein signal transduction 65 28 4,79 1,37164018691589E-08
GO:0035567 non-canonical Wnt signaling pathway 93 40 6,86 0,000270535483871
GO:0048193 Golgi vesicle transport 320 133 23,59 8,53760526315789E-15
GO:0098693 regulation of synaptic vesicle cycle 53 22 3,91 0,000382866521739
GO:0017156 calcium ion regulated exocytosis 94 38 6,93 1,7931875E-08
GO:0033059 cellular pigmentation 47 19 3,47 3,5349406779661E-08
GO:0016482 cytosolic transport 132 53 9,73 6,3149197080292E-07
GO:0001738 morphogenesis of a polarized epithelium 82 31 6,05 1,04900617283951E-05
GO:0008088 axo-dendritic transport 53 20 3,91 0,000134991262136
GO:0099643 signal release from synapse 116 43 8,55 1,60927083333333E-09
GO:0007269 neurotransmitter secretion 116 43 8,55 1,38763473053892E-05
GO:0019882 antigen processing and presentation 162 58 11,94 1,02120508474576E-11
GO:0033619 membrane protein proteolysis 54 19 3,98 5,020925E-08
GO:0099531 presynaptic process involved in chemical... 123 43 9,07 3,65158181818182E-16
GO:0042108 positive regulation of cytokine biosynth... 52 18 3,83 0,0003034625
GO:1905330 regulation of morphogenesis of an epithe... 111 38 8,18 3,74521212121212E-07
GO:0007030 Golgi organization 113 38 8,33 1,58502727272727E-10
GO:0043473 pigmentation 85 28 6,27 8,63815053763441E-10
GO:0016197 endosomal transport 174 56 12,83 2,54653846153846E-05
GO:0042089 cytokine biosynthetic process 95 29 7 4,79451724137931E-10
GO:0007034 vacuolar transport 118 36 8,7 2,70146448087432E-05
GO:0042107 cytokine metabolic process 96 29 7,08 7,7245E-08
GO:0031341 regulation of cell killing 70 21 5,16 1,13734969325153E-05
GO:0032526 response to retinoic acid 87 26 6,41 0,00069933744856
GO:0035384 thioester biosynthetic process 44 13 3,24 0,000161846666667
GO:0070304 positive regulation of stress-activated ... 144 42 10,62 0,000315846222222
GO:0042035 regulation of cytokine biosynthetic proc... 86 25 6,34 1,931125E-06
GO:0050435 amyloid-beta metabolic process 35 10 2,58 7,01510714285714E-05
GO:0061024 membrane organization 668 190 49,25 2,207E-27
GO:0045995 regulation of embryonic development 103 29 7,59 0,000412861206897
GO:0032872 regulation of stress-activated MAPK casc... 199 56 14,67 0,00028661040724
GO:0070302 regulation of stress-activated protein k... 201 56 14,82 2,68678260869565E-08
GO:0022406 membrane docking 159 44 11,72 2,03125740740741E-12
GO:0051091 positive regulation of DNA binding trans... 228 63 16,81 1,03837540983607E-11
GO:2000027 regulation of organ morphogenesis 171 47 12,61 2,5346015625E-07
GO:0051403 stress-activated MAPK cascade 241 66 17,77 9,17762376237624E-05
GO:0072659 protein localization to plasma membrane 194 53 14,3 3,20863846153846E-07
GO:0043406 positive regulation of MAP kinase activi... 217 59 16 0,000297895515695
GO:0051656 establishment of organelle localization 398 107 29,34 1,02993333333333E-13
GO:0043405 regulation of MAP kinase activity 283 76 20,87 2,60032673267327E-09
GO:0060627 regulation of vesicle-mediated transport 391 103 28,83 4,55338947368421E-20
GO:0031098 stress-activated protein kinase signalin... 265 69 19,54 1,359512E-10
GO:1905475 regulation of protein localization to me... 149 37 10,99 0,000638388429752
GO:0198738 cell-cell signaling by wnt 400 98 29,49 6,94085507246377E-07
GO:0010256 endomembrane system organization 335 80 24,7 5,25266E-13
GO:0051640 organelle localization 565 134 41,66 9,20365957446809E-07
5 GO:0030100 regulation of endocytosis 216 51 15,93 0,000923247011952
GO:0019218 regulation of steroid metabolic process 102 24 7,52 0,000250331018519
GO:0045165 cell fate commitment 203 47 14,97 2,38990055248619E-05
GO:0005976 polysaccharide metabolic process 92 21 6,78 0,000168401421801
GO:0016192 vesicle-mediated transport 1644 375 121,21 2,207E-27
GO:1905114 cell surface receptor signaling pathway ... 482 107 35,54 4,69191851851852E-07
GO:0006470 protein dephosphorylation 263 58 19,39 1,5449E-08
GO:0001819 positive regulation of cytokine producti... 357 78 26,32 0,000813105263158
GO:0001505 regulation of neurotransmitter levels 263 57 19,39 1,73686982248521E-05
GO:0051090 regulation of DNA binding transcription ... 360 77 26,54 2,07242682926829E-10
GO:0034219 carbohydrate transmembrane transport 99 21 7,3 0,00033348061674
GO:0001906 cell killing 138 29 10,17 1,96461271676301E-05
GO:0035725 sodium ion transmembrane transport 119 25 8,77 2,73247619047619E-06
GO:0050867 positive regulation of cell activation 274 55 20,2 7,2147614213198E-05
GO:0098657 import into cell 650 128 47,92 7,67529936305733E-06
GO:0002521 leukocyte differentiation 430 84 31,7 2,53956164383562E-06
GO:0046907 intracellular transport 1531 297 112,88 2,02025384615385E-21
GO:1903532 positive regulation of secretion by cell 320 60 23,59 0,000479905106383
GO:0043068 positive regulation of programmed cell d... 518 97 38,19 5,793375E-06
GO:0043065 positive regulation of apoptotic process 514 96 37,9 5,20474331550802E-05
GO:0010942 positive regulation of cell death 562 104 41,44 2,04935714285714E-09
GO:0051649 establishment of localization in cell 1838 335 135,51 5,2968E-21
GO:0046890 regulation of lipid biosynthetic process 156 28 11,5 0,00059616473029
GO:1903530 regulation of secretion by cell 578 103 42,61 5,59746376811594E-11
GO:0001816 cytokine production 601 107 44,31 1,12129838709677E-11
GO:0000165 MAPK cascade 773 136 56,99 0,000113041463415
GO:0034097 response to cytokine 970 169 71,52 1,22461585365854E-05
GO:0043408 regulation of MAPK cascade 632 110 46,6 0,000345569736842
GO:0045597 positive regulation of cell differentiat... 760 132 56,03 3,68931343283582E-11
GO:0023014 signal transduction by protein phosphory... 789 137 58,17 4,08944117647059E-09
GO:0015833 peptide transport 1701 287 125,41 1,19511132075472E-08
GO:0045321 leukocyte activation 1016 171 74,91 0,00033348061674
GO:0042886 amide transport 1719 288 126,74 1,81752941176471E-05
GO:0050865 regulation of cell activation 436 73 32,15 1,08034965034965E-06
GO:0045184 establishment of protein localization 1760 294 129,76 7,0624E-11
GO:0080135 regulation of cellular response to stres... 561 93 41,36 4,94967961165049E-09
GO:0030182 neuron differentiation 1051 169 77,49 1,71655555555556E-07
GO:0031175 neuron projection development 762 121 56,18 1,5449E-08
GO:0051641 cellular localization 2367 374 174,51 1,3468358974359E-14
GO:0051049 regulation of transport 1476 233 108,82 0,000161846666667
GO:0016051 carbohydrate biosynthetic process 184 29 13,57 0,000592211666667
GO:0005996 monosaccharide metabolic process 255 40 18,8 7,76331658291457E-05
GO:0001775 cell activation 1152 180 84,93 1,25523125E-20
GO:0032940 secretion by cell 1227 191 90,46 7,338275E-22
GO:0007267 cell-cell signaling 1315 204 96,95 5,33899264705882E-07
GO:0002684 positive regulation of immune system pro... 814 126 60,01 4,06989784946237E-05
GO:0051050 positive regulation of transport 808 125 59,57 0,000813105263158
GO:0071705 nitrogen compound transport 1944 299 143,33 0,000514966666667
GO:0030155 regulation of cell adhesion 552 84 40,7 2,34398620689655E-06
GO:0008104 protein localization 2255 341 166,26 0,000430980686695
GO:0048534 hematopoietic or lymphoid organ developm... 716 107 52,79 1,89724561403509E-05
GO:0051094 positive regulation of developmental pro... 1072 160 79,04 2,18470707070707E-09
GO:0022008 neurogenesis 1241 183 91,5 9,24764084507042E-07
GO:0045937 positive regulation of phosphate metabol... 954 140 70,34 0,0003508069869
GO:0051240 positive regulation of multicellular org... 1315 192 96,95 7,35666666666667E-14
GO:0033036 macromolecule localization 2514 356 185,35 5,95745751633987E-06
GO:0043067 regulation of programmed cell death 1277 177 94,15 0,000923247011952
GO:0007166 cell surface receptor signaling pathway 2446 337 180,34 1,00847638888889E-10
GO:0002682 regulation of immune system process 1169 161 86,19 2,47184E-09
GO:1901700 response to oxygen-containing compound 1341 184 98,87 0,000161846666667
GO:0080134 regulation of response to stress 1136 155 83,75 0,000756685714286
GO:2000026 regulation of multicellular organismal d... 1566 213 115,46 2,5346015625E-07
5 GO:0045595 regulation of cell differentiation 1377 186 101,52 1,87319125E-10
GO:0002376 immune system process 2380 319 175,47 2,207E-27
GO:0051707 response to other organism 763 102 56,25 5,85435789473684E-05
GO:0010648 negative regulation of cell communicatio... 1102 147 81,25 0,000292278378378
GO:0023057 negative regulation of signaling 1106 147 81,54 6,97185641025641E-05
GO:0060548 negative regulation of cell death 828 110 61,05 0,000275119178082
GO:0006914 autophagy 392 52 28,9 0,000174073239437
GO:0045944 positive regulation of transcription by ... 989 130 72,92 9,79161971830986E-11
GO:0005975 carbohydrate metabolic process 518 68 38,19 8,50672784810127E-06
GO:0070887 cellular response to chemical stimulus 2671 346 196,93 8,98197674418605E-14
GO:0050793 regulation of developmental process 1985 257 146,35 2,74648888888889E-15
GO:0010033 response to organic substance 2704 348 199,36 6,39268965517241E-12
GO:0035556 intracellular signal transduction 2361 303 174,07 6,22813089005236E-05
GO:0051234 establishment of localization 4186 533 308,63 5,25923404255319E-05
GO:0051241 negative regulation of multicellular org... 919 116 67,76 1,65525E-05
GO:0045893 positive regulation of transcription, DN... 1279 160 94,3 2,10668181818182E-11
GO:0048585 negative regulation of response to stimu... 1294 160 95,4 7,62546794871795E-06
GO:0042127 regulation of cell proliferation 1301 156 95,92 0,000113041463415
GO:0008219 cell death 1804 215 133,01 2,08438888888889E-11
GO:1903508 positive regulation of nucleic acid-temp... 1357 160 100,05 4,36207058823529E-10
GO:1902680 positive regulation of RNA biosynthetic ... 1358 160 100,12 1,25262162162162E-10
GO:0009605 response to external stimulus 1929 226 142,22 0,00048441779661
GO:0040007 growth 764 89 56,33 0,000562369456067
GO:0019362 pyridine nucleotide metabolic process 129 15 9,51 0,00028661040724
GO:0010628 positive regulation of gene expression 1621 188 119,51 2,03125740740741E-12
GO:0051254 positive regulation of RNA metabolic pro... 1428 162 105,28 2,04935714285714E-09
GO:0048869 cellular developmental process 3276 363 241,53 5,685232E-17
GO:0023052 signaling 5319 589 392,16 2,207E-27
GO:0010557 positive regulation of macromolecule bio... 1575 174 116,12 3,03698290598291E-08
GO:0009891 positive regulation of biosynthetic proc... 1683 184 124,08 5,61781818181818E-10
GO:0031328 positive regulation of cellular biosynth... 1655 180 122,02 5,45258823529412E-11
GO:0042221 response to chemical 3908 411 288,13 0,000872120967742
GO:0045935 positive regulation of nucleobase-contai... 1588 167 117,08 3,67833333333333E-10
5 GO:0016043 cellular component organization 5129 496 378,15 3,54039583333333E-13
GO:0048523 negative regulation of cellular process 3779 342 278,62 0,000250331018519
GO:0006355 regulation of transcription, DNA-templat... 2729 233 201,2 0,000538767647059
GO:2001141 regulation of RNA biosynthetic process 2778 233 204,82 0,000923247011952
GO:0080090 regulation of primary metabolic process 4805 397 354,26 7,00206474820144E-07
GO:0060255 regulation of macromolecule metabolic pr... 4858 397 358,17 9,17762376237624E-05
GO:0006351 transcription, DNA-templated 2873 234 211,82 2,75447286821705E-07
GO:0051171 regulation of nitrogen compound metaboli... 4663 376 343,79 2,00749717514124E-05
GO:0032774 RNA biosynthetic process 2917 234 215,06 1,5835225E-14
GO:2000112 regulation of cellular macromolecule bio... 3101 245 228,63 3,0898E-05
GO:0044260 cellular macromolecule metabolic process 6674 511 492,06 9,13337106918239E-06
GO:0034654 nucleobase-containing compound biosynthe... 3345 244 246,62 2,95831914893617E-13
GO:0019438 aromatic compound biosynthetic process 3406 248 251,12 5,05063461538462E-09
GO:1901362 organic cyclic compound biosynthetic pro... 3535 256 260,63 2,79680172413793E-08
GO:0018130 heterocycle biosynthetic process 3399 246 250,6 4,28417647058823E-08
GO:1901030 positive regulation of mitochondrial out... 31 11 0,49 3,70776E-07
GO:0051204 protein insertion into mitochondrial mem... 31 10 0,49 8,36820833333333E-07
GO:1901028 regulation of mitochondrial outer membra... 37 11 0,58 6,67115909090909E-07
GO:1902686 mitochondrial outer membrane permeabiliz... 49 13 0,77 3,83884242424242E-06
GO:0051205 protein insertion into membrane 43 11 0,68 3,83884242424242E-06
GO:0003197 endocardial cushion development 40 10 0,63 0,000120158888889
GO:0060389 pathway-restricted SMAD protein phosphor... 59 14 0,93 3,62252413793103E-06
GO:0060393 regulation of pathway-restricted SMAD pr... 56 13 0,88 2,2787275E-05
GO:0046902 regulation of mitochondrial membrane per... 59 13 0,93 3,73766129032258E-06
GO:0006099 tricarboxylic acid cycle 32 7 0,51 0,000205986666667
6 GO:0090559 regulation of membrane permeability 65 13 1,03 4,414E-07
GO:0097711 ciliary basal body-plasma membrane docki... 86 16 1,36 0,00027409516129
GO:0060411 cardiac septum morphogenesis 64 11 1,01 6,11522916666667E-05
GO:0008637 apoptotic mitochondrial changes 102 16 1,61 3,26145555555556E-05
GO:0010822 positive regulation of mitochondrion org... 100 15 1,58 4,31663235294118E-06
GO:2001235 positive regulation of apoptotic signali... 142 21 2,24 9,57838E-05
GO:0031929 TOR signaling 94 13 1,49 0,000952335616438
GO:0003279 cardiac septum development 87 12 1,37 4,60182978723404E-05
GO:0003206 cardiac chamber morphogenesis 106 13 1,67 0,000201622542373
GO:0030509 BMP signaling pathway 123 15 1,94 3,62252413793103E-06
GO:0007006 mitochondrial membrane organization 111 13 1,75 0,000613416176471
GO:0022406 membrane docking 159 17 2,51 8,65144E-07
GO:0003205 cardiac chamber development 133 14 2,1 0,000109925576923
GO:0046661 male sex differentiation 131 13 2,07 0,000944105555556
GO:2001233 regulation of apoptotic signaling pathwa... 334 31 5,28 5,57880555555556E-06
GO:0000226 microtubule cytoskeleton organization 436 39 6,89 0,000120158888889
GO:0007017 microtubule-based process 602 51 9,51 2,62633E-18
GO:0003007 heart morphogenesis 210 17 3,32 0,000761570422535
GO:0097190 apoptotic signaling pathway 496 38 7,84 7,7245E-09
GO:0010942 positive regulation of cell death 562 37 8,88 3,35847826086957E-05
GO:0007389 pattern specification process 349 20 5,51 0,000613416176471
GO:0022402 cell cycle process 1118 51 17,66 3,0898E-08
GO:0051726 regulation of cell cycle 983 44 15,53 2,90139756097561E-05
GO:0007049 cell cycle 1522 66 24,05 3,028004E-10
GO:0009719 response to endogenous stimulus 1405 56 22,2 3,65626333333333E-06
GO:0031399 regulation of protein modification proce... 1515 60 23,94 0,000427818461538
GO:0051240 positive regulation of multicellular org... 1315 51 20,78 0,000109051764706
GO:0006468 protein phosphorylation 1629 61 25,74 0,000123592
GO:0008219 cell death 1804 67 28,5 8,23946666666667E-08
6 GO:0033554 cellular response to stress 1625 59 25,67 0,000192434912281
GO:0045595 regulation of cell differentiation 1377 48 21,76 0,00075038
GO:0009966 regulation of signal transduction 2612 89 41,27 6,26310810810811E-06
GO:0070887 cellular response to chemical stimulus 2671 85 42,2 3,16165581395349E-05
GO:0051239 regulation of multicellular organismal p... 2354 68 37,19 0,000694085507246
GO:0006793 phosphorus metabolic process 2768 76 43,73 0,0003620859375
GO:0031323 regulation of cellular metabolic process 4883 120 77,15 1,06954615384615E-06
GO:0007165 signal transduction 4969 118 78,5 0,000253262295082
GO:0051171 regulation of nitrogen compound metaboli... 4663 110 73,67 0,0001710425
GO:0019222 regulation of metabolic process 5251 123 82,96 8,36820833333333E-07
GO:0060255 regulation of macromolecule metabolic pr... 4858 111 76,75 0,000318788888889
GO:0023052 signaling 5319 119 84,03 0,000201622542373
GO:0071840 cellular component organization or bioge... 5277 117 83,37 0,000444743939394
GO:0046755 viral budding 23 23 3,78 1,53466887417219E-11
GO:0006293 nucleotide-excision repair, preincision ... 20 20 3,29 1,58082790697674E-07
GO:0001325 formation of extrachromosomal circular D... 16 16 2,63 2,0741712962963E-07
GO:0043248 proteasome assembly 14 14 2,3 4,77514545454545E-07
GO:1904896 ESCRT complex disassembly 10 10 1,64 1,3923991416309E-06
GO:0061641 CENP-A containing chromatin organization 34 34 5,59 3,59146494464945E-05
GO:1901409 positive regulation of phosphorylation o... 7 7 1,15 6,59743772241993E-05
GO:0006297 nucleotide-excision repair, DNA gap fill... 23 23 3,78 0,000243408681672
GO:1904429 regulation of t-circle formation 8 8 1,31 0,000257483333333
7 GO:1902047 polyamine transmembrane transport 4 4 0,66 0,000432572
GO:0015846 polyamine transport 5 5 0,82 0,000603114114114
GO:1900264 positive regulation of DNA-directed DNA ... 7 7 1,15 0,000887873563218
GO:0000731 DNA synthesis involved in DNA repair 47 46 7,72 1,689734375E-05
GO:0034724 DNA replication-independent nucleosome o... 45 44 7,39 0,000681573529412
GO:0033683 nucleotide-excision repair, DNA incision 38 37 6,24 2,17591549295775E-12
GO:0036258 multivesicular body assembly 29 28 4,76 7,24171875E-14
GO:0006270 DNA replication initiation 29 28 4,76 0,000209986407767
GO:0032201 telomere maintenance via semi-conservati... 24 23 3,94 6,4918670212766E-09
GO:0061952 midbody abscission 16 15 2,63 8,77784090909091E-10
GO:0036257 multivesicular body organization 30 28 4,93 3,13301398601399E-12
GO:0007130 synaptonemal complex assembly 15 14 2,46 1,01638157894737E-06
GO:1904837 beta-catenin-TCF complex assembly 28 26 4,6 3,63505882352941E-13
GO:0007131 reciprocal meiotic recombination 45 41 7,39 0,000143235761589
GO:0043562 cellular response to nitrogen levels 10 9 1,64 0,000115999317406
GO:0006995 cellular response to nitrogen starvation 10 9 1,64 0,00030898
GO:0031441 negative regulation of mRNA 3'-end proce... 9 8 1,48 1,73651550387597E-05
GO:0034243 regulation of transcription elongation f... 26 23 4,27 8,17014423076923E-08
GO:0032968 positive regulation of transcription elo... 17 15 2,79 0,000643708333333
GO:0070816 phosphorylation of RNA polymerase II C-t... 16 14 2,63 1,57519215686275E-05
GO:0051984 positive regulation of chromosome segreg... 26 22 4,27 1,32642929292929E-08
GO:0051782 negative regulation of cell division 13 11 2,14 0,000853287790698
GO:0051383 kinetochore organization 19 16 3,12 0,000425684520124
GO:0065004 protein-DNA complex assembly 189 156 31,05 2,37676923076923E-28
GO:0045023 G0 to G1 transition 40 33 6,57 0,000335847826087
GO:0071824 protein-DNA complex subunit organization 226 185 37,13 2,37676923076923E-28
GO:0001711 endodermal cell fate commitment 11 9 1,81 0,000887873563218
GO:0006338 chromatin remodeling 139 113 22,84 0,000681573529412
GO:0044786 cell cycle DNA replication 58 47 9,53 1,39892519685039E-05
GO:0000209 protein polyubiquitination 231 187 37,95 1,64905056179775E-09
GO:1903513 endoplasmic reticulum to cytosol transpo... 26 21 4,27 1,06138167938931E-13
GO:0006368 transcription elongation from RNA polyme... 72 58 11,83 7,1824298245614E-17
GO:0097329 response to antimetabolite 10 8 1,64 0,000469574468085
GO:0031497 chromatin assembly 120 95 19,72 2,37676923076923E-28
GO:0061982 meiosis I cell cycle process 95 75 15,61 0,000129605704698
GO:0008156 negative regulation of DNA replication 38 30 6,24 0,000185588636364
GO:0051095 regulation of helicase activity 14 11 2,3 1,27069696969697E-06
GO:0051304 chromosome separation 83 65 13,64 5,95791031390134E-07
GO:0061418 regulation of transcription from RNA pol... 27 21 4,44 7,09671378091873E-05
GO:0051096 positive regulation of helicase activity 9 7 1,48 0,000101217586207
GO:0000083 regulation of transcription involved in ... 27 21 4,44 0,000139041
GO:0010992 ubiquitin recycling 9 7 1,48 0,000151957377049
GO:0009396 folic acid-containing compound biosynthe... 9 7 1,48 0,000283557594937
GO:0019068 virion assembly 35 27 5,75 1,24470616113744E-07
GO:1902188 positive regulation of viral release fro... 13 10 2,14 4,55872131147541E-06
GO:0006281 DNA repair 470 361 77,22 2,37676923076923E-28
7 GO:0042769 DNA damage response, detection of DNA da... 38 29 6,24 1,78945945945946E-05
GO:0045814 negative regulation of gene expression, ... 97 74 15,94 5,39965048543689E-19
GO:0006342 chromatin silencing 80 60 13,14 4,34104132231405E-06
GO:0051457 maintenance of protein location in nucle... 20 15 3,29 3,17050373134328E-05
GO:0006363 termination of RNA polymerase I transcri... 27 20 4,44 3,63505882352941E-05
GO:0033045 regulation of sister chromatid segregati... 77 57 12,65 0,000643708333333
GO:0051985 negative regulation of chromosome segreg... 44 32 7,23 1,18838461538462E-08
GO:0006354 DNA-templated transcription, elongation 84 61 13,8 1,94633070866142E-14
GO:0034502 protein localization to chromosome 77 55 12,65 2,86682474226804E-20
GO:0036260 RNA capping 30 21 4,93 1,87595E-12
GO:0051443 positive regulation of ubiquitin-protein... 30 21 4,93 3,81376987447699E-06
GO:0032509 endosome transport via multivesicular bo... 23 16 3,78 0,000266532268371
GO:0006260 DNA replication 243 169 39,92 2,37676923076923E-28
GO:0000070 mitotic sister chromatid segregation 142 98 23,33 0,0009655625
GO:0006370 7-methylguanosine mRNA capping 29 20 4,76 2,92824183006536E-11
GO:0098813 nuclear chromosome segregation 227 156 37,29 6,50484210526316E-10
GO:0051983 regulation of chromosome segregation 98 67 16,1 6,82163636363636E-24
GO:0032784 regulation of DNA-templated transcriptio... 41 28 6,74 2,05595057034221E-05
GO:0070646 protein modification by small protein re... 258 175 42,39 4,58376923076923E-21
GO:0010498 proteasomal protein catabolic process 407 275 66,87 5,5175E-17
GO:1904029 regulation of cyclin-dependent protein k... 92 62 15,11 8,09233333333333E-09
GO:0000079 regulation of cyclin-dependent protein s... 91 61 14,95 3,94807777777778E-21
GO:0019941 modification-dependent protein catabolic... 539 361 88,55 3,00954545454545E-11
GO:0006325 chromatin organization 667 444 109,58 2,37676923076923E-28
GO:0007059 chromosome segregation 269 179 44,19 2,37676923076923E-28
GO:0016570 histone modification 403 268 66,21 2,37676923076923E-28
GO:0016569 covalent chromatin modification 413 274 67,85 2,37676923076923E-28
GO:0051276 chromosome organization 1027 679 168,73 2,37676923076923E-28
GO:0006367 transcription initiation from RNA polyme... 165 109 27,11 2,99521428571429E-20
GO:0043632 modification-dependent macromolecule cat... 549 361 90,2 2,37676923076923E-28
GO:0070647 protein modification by small protein co... 904 591 148,52 2,37676923076923E-28
GO:0098534 centriole assembly 34 22 5,59 0,000270603503185
GO:0006275 regulation of DNA replication 96 62 15,77 3,0898E-10
GO:0010948 negative regulation of cell cycle proces... 256 165 42,06 1,99837810945274E-08
GO:1903046 meiotic cell cycle process 157 101 25,79 1,03745109489051E-12
GO:0035601 protein deacylation 93 59 15,28 1,32642929292929E-08
GO:0098732 macromolecule deacylation 94 59 15,44 8,85098958333333E-09
GO:0000075 cell cycle checkpoint 204 127 33,52 2,37676923076923E-28
GO:0051603 proteolysis involved in cellular protein... 618 384 101,53 2,37676923076923E-28
GO:0006974 cellular response to DNA damage stimulus 724 449 118,95 2,37676923076923E-28
GO:0007214 gamma-aminobutyric acid signaling pathwa... 21 13 3,45 1,95994776119403E-13
GO:2001022 positive regulation of response to DNA d... 76 47 12,49 8,21004E-10
GO:0006352 DNA-templated transcription, initiation 211 130 34,67 1,66920229885057E-21
GO:0071985 multivesicular body sorting pathway 31 19 5,09 1,34789932885906E-11
GO:0031440 regulation of mRNA 3'-end processing 23 14 3,78 4,03348275862069E-08
GO:0036503 ERAD pathway 79 48 12,98 1,01638157894737E-06
GO:0051321 meiotic cell cycle 211 128 34,67 2,97536296296296E-13
GO:1904031 positive regulation of cyclin-dependent ... 33 20 5,42 0,000106178694158
GO:0043687 post-translational protein modification 320 192 52,57 8,36820833333333E-26
GO:0045652 regulation of megakaryocyte differentiat... 60 36 9,86 4,23871480144404E-05
GO:0045324 late endosome to vacuole transport 15 9 2,46 0,000151957377049
GO:0044770 cell cycle phase transition 477 285 78,37 6,06331720430108E-09
GO:0045815 positive regulation of gene expression, ... 52 31 8,54 0,000887873563218
GO:0006259 DNA metabolic process 858 509 140,96 2,37676923076923E-28
GO:0051783 regulation of nuclear division 190 112 31,22 6,77216438356165E-12
GO:0018205 peptidyl-lysine modification 306 180 50,27 2,37676923076923E-28
GO:0044257 cellular protein catabolic process 664 390 109,09 2,37676923076923E-28
GO:0051053 negative regulation of DNA metabolic pro... 130 76 21,36 6,896875E-07
GO:0072395 signal transduction involved in cell cyc... 69 40 11,34 7,4617619047619E-12
GO:1990182 exosomal secretion 19 11 3,12 8,92608888888889E-07
GO:0006473 protein acetylation 170 98 27,93 2,55547368421053E-05
GO:0031058 positive regulation of histone modificat... 77 44 12,65 8,58277777777778E-05
GO:0032922 circadian regulation of gene expression 51 29 8,38 8,85098958333333E-09
GO:0048285 organelle fission 401 228 65,88 3,04298484848485E-28
GO:0051303 establishment of chromosome localization 67 38 11,01 1,29823529411765E-16
GO:0071897 DNA biosynthetic process 166 94 27,27 9,62591538461539E-14
GO:0010212 response to ionizing radiation 131 74 21,52 8,06574297188755E-06
GO:0072595 maintenance of protein localization in o... 32 18 5,26 0,000724879765396
GO:0009411 response to UV 121 68 19,88 6,36135294117647E-10
GO:0050000 chromosome localization 68 38 11,17 7,09576162790698E-10
GO:0060968 regulation of gene silencing 89 49 14,62 1,97221276595745E-12
GO:1901796 regulation of signal transduction by p53... 120 66 19,72 2,82603658536585E-22
7 GO:0009147 pyrimidine nucleoside triphosphate metab... 20 11 3,29 0,000681573529412
GO:0040029 regulation of gene expression, epigeneti... 235 129 38,61 3,27155294117647E-22
GO:0009649 entrainment of circadian clock 24 13 3,94 4,26370250896057E-05
GO:0008213 protein alkylation 150 81 24,64 4,09151598173516E-07
GO:0006479 protein methylation 150 81 24,64 9,23247011952191E-06
GO:0032527 protein exit from endoplasmic reticulum 39 21 6,41 1,17706666666667E-07
GO:1902186 regulation of viral release from host ce... 26 14 4,27 3,44587360594796E-05
GO:0051054 positive regulation of DNA metabolic pro... 186 100 30,56 1,31170754716981E-07
GO:0031056 regulation of histone modification 123 66 20,21 2,74939830508475E-06
GO:0030219 megakaryocyte differentiation 73 39 11,99 1,84859829059829E-06
GO:0051052 regulation of DNA metabolic process 348 185 57,17 8,74088157894737E-25
GO:1903047 mitotic cell cycle process 716 379 117,63 0,000120449830508
GO:0030163 protein catabolic process 792 414 130,12 2,37676923076923E-28
GO:0042770 signal transduction in response to DNA d... 123 64 20,21 3,07272928176796E-09
GO:0072331 signal transduction by p53 class mediato... 200 104 32,86 2,37676923076923E-28
GO:0060964 regulation of gene silencing by miRNA 56 29 9,2 1,8631959798995E-08
GO:0002200 somatic diversification of immune recept... 58 30 9,53 2,08514110429448E-10
GO:0035329 hippo signaling 33 17 5,42 0,000115999317406
GO:0043543 protein acylation 193 99 31,71 3,06132258064516E-07
GO:0031023 microtubule organizing center organizati... 123 63 20,21 4,71795808383234E-10
GO:0001824 blastocyst development 49 25 8,05 7,50558704453441E-06
GO:0061726 mitochondrion disassembly 64 32 10,51 5,84556756756757E-09
GO:0052126 movement in host environment 30 15 4,93 3,59146494464945E-05
GO:0006760 folic acid-containing compound metabolic... 24 12 3,94 4,10100727272727E-05
GO:0052192 movement in environment of other organis... 30 15 4,93 0,000887873563218
GO:0000278 mitotic cell cycle 835 417 137,18 0,00044409969419
GO:0051225 spindle assembly 91 45 14,95 0,000166606862745
GO:1903362 regulation of cellular protein catabolic... 209 103 34,34 6,12475147928994E-10
GO:0045786 negative regulation of cell cycle 495 242 81,33 6,7589375E-23
GO:0000045 autophagosome assembly 78 38 12,81 0,000603114114114
GO:0043401 steroid hormone mediated signaling pathw... 177 86 29,08 7,15929268292683E-08
GO:0022402 cell cycle process 1118 543 183,68 2,37676923076923E-28
GO:0045732 positive regulation of protein catabolic... 183 88 30,07 2,49813617021277E-06
GO:0016458 gene silencing 186 89 30,56 6,95205E-20
GO:0051301 cell division 538 257 88,39 2,37676923076923E-28
GO:0032204 regulation of telomere maintenance 73 34 11,99 1,07940611353712E-06
GO:0019827 stem cell population maintenance 125 58 20,54 3,213392E-15
GO:0042176 regulation of protein catabolic process 319 148 52,41 1,6385303030303E-13
GO:0045747 positive regulation of Notch signaling p... 41 19 6,74 0,000296348742138
GO:0098727 maintenance of cell number 128 59 21,03 1,53466887417219E-11
GO:0070507 regulation of microtubule cytoskeleton o... 154 70 25,3 3,86225E-12
GO:0051726 regulation of cell cycle 983 444 161,5 2,37676923076923E-28
GO:0007049 cell cycle 1522 684 250,05 2,37676923076923E-28
GO:0007032 endosome organization 67 30 11,01 1,11487628865979E-08
GO:0033143 regulation of intracellular steroid horm... 72 32 11,83 6,90808943089431E-06
GO:0097711 ciliary basal body-plasma membrane docki... 86 38 14,13 1,16596226415094E-10
GO:0007569 cell aging 85 37 13,96 1,27069696969697E-06
GO:0044265 cellular macromolecule catabolic process 992 430 162,98 2,37676923076923E-28
GO:0035195 gene silencing by miRNA 86 37 14,13 5,48597142857143E-06
GO:0006998 nuclear envelope organization 49 21 8,05 2,05595057034221E-05
GO:0045787 positive regulation of cell cycle 349 149 57,34 7,23334682080925E-10
GO:0030522 intracellular receptor signaling pathway 257 107 42,22 3,12658333333333E-22
GO:0009755 hormone-mediated signaling pathway 217 90 35,65 4,53548623853211E-18
GO:0043414 macromolecule methylation 249 102 40,91 1,22126482213439E-05
GO:0032886 regulation of microtubule-based process 177 72 29,08 8,21004E-10
GO:0010639 negative regulation of organelle organiz... 327 133 53,72 0,00044409969419
GO:0071383 cellular response to steroid hormone sti... 227 92 37,29 1,04313760683761E-16
GO:0000122 negative regulation of transcription by ... 689 278 113,2 8,17014423076923E-08
GO:0045892 negative regulation of transcription, DN... 970 389 159,36 6,45337974683544E-23
GO:1903507 negative regulation of nucleic acid-temp... 1008 400 165,61 6,7669173553719E-16
GO:1902679 negative regulation of RNA biosynthetic ... 1009 400 165,77 8,3866E-19
GO:0033554 cellular response to stress 1625 643 266,98 2,37676923076923E-28
GO:1901565 organonitrogen compound catabolic proces... 1107 438 181,87 2,37676923076923E-28
GO:0030178 negative regulation of Wnt signaling pat... 139 54 22,84 0,000469574468085
GO:0045934 negative regulation of nucleobase-contai... 1206 467 198,14 2,37676923076923E-28
GO:0009057 macromolecule catabolic process 1189 458 195,34 2,37676923076923E-28
GO:0009314 response to radiation 384 147 63,09 4,4851935483871E-11
GO:2000113 negative regulation of cellular macromol... 1173 447 192,72 1,5449E-26
GO:0051253 negative regulation of RNA metabolic pro... 1080 409 177,44 2,37676923076923E-28
GO:0010558 negative regulation of macromolecule bio... 1234 461 202,74 2,37676923076923E-28
GO:0002312 B cell activation involved in immune res... 59 22 9,69 0,000810734693878
GO:0042752 regulation of circadian rhythm 89 33 14,62 0,0009655625
7 GO:0030111 regulation of Wnt signaling pathway 262 97 43,04 1,07471304347826E-12
GO:0007224 smoothened signaling pathway 106 39 17,42 1,90141538461538E-05
GO:0032259 methylation 294 108 48,3 1,26114285714286E-08
GO:0030177 positive regulation of Wnt signaling pat... 109 40 17,91 0,000327268535826
GO:0031647 regulation of protein stability 239 87 39,27 1,73259813084112E-18
GO:0045935 positive regulation of nucleobase-contai... 1588 571 260,9 2,37676923076923E-28
GO:0016055 Wnt signaling pathway 398 143 65,39 1,91913043478261E-10
GO:0198738 cell-cell signaling by wnt 400 143 65,72 2,37676923076923E-28
GO:0031327 negative regulation of cellular biosynth... 1299 463 213,42 2,37676923076923E-28
GO:1903508 positive regulation of nucleic acid-temp... 1357 483 222,95 1,64905056179775E-09
GO:0046785 microtubule polymerization 59 21 9,69 8,06574297188755E-06
GO:1902680 positive regulation of RNA biosynthetic ... 1358 483 223,11 1,85852631578947E-13
GO:0051932 synaptic transmission, GABAergic 34 12 5,59 8,58277777777778E-05
GO:0009890 negative regulation of biosynthetic proc... 1319 464 216,7 2,37676923076923E-28
GO:0045944 positive regulation of transcription by ... 989 347 162,49 0,00012484040404
GO:0016073 snRNA metabolic process 83 29 13,64 4,10263900414938E-06
GO:0006508 proteolysis 1485 515 243,98 1,95145263157895E-20
GO:0045637 regulation of myeloid cell differentiati... 199 69 32,69 3,12451685393258E-05
GO:0051254 positive regulation of RNA metabolic pro... 1428 494 234,61 8,3009552238806E-28
GO:0071453 cellular response to oxygen levels 148 51 24,32 2,01251340996169E-05
GO:0045893 positive regulation of transcription, DN... 1279 439 210,13 5,54148913043478E-09
GO:0002712 regulation of B cell mediated immunity 44 15 7,23 0,000185588636364
GO:0000226 microtubule cytoskeleton organization 436 148 71,63 2,37676923076923E-28
GO:0002377 immunoglobulin production 80 27 13,14 7,58898245614035E-05
GO:0051099 positive regulation of binding 163 55 26,78 0,000143235761589
GO:0010557 positive regulation of macromolecule bio... 1575 528 258,76 4,67062790697674E-22
GO:0032606 type I interferon production 99 33 16,27 1,03486124401914E-07
GO:0080135 regulation of cellular response to stres... 561 185 92,17 1,22258273381295E-12
GO:0051098 regulation of binding 325 107 53,4 5,793375E-10
GO:0032479 regulation of type I interferon producti... 98 32 16,1 3,11814678899083E-07
GO:0031328 positive regulation of cellular biosynth... 1655 531 271,91 8,88846575342466E-26
GO:0009891 positive regulation of biosynthetic proc... 1683 535 276,51 3,63505882352941E-27
GO:0006464 cellular protein modification process 3414 1081 560,9 2,37676923076923E-28
GO:0007219 Notch signaling pathway 156 49 25,63 0,00012484040404
GO:0097193 intrinsic apoptotic signaling pathway 244 76 40,09 8,17014423076923E-08
GO:0006996 organelle organization 3158 981 518,84 2,37676923076923E-28
GO:0051172 negative regulation of nitrogen compound... 1946 603 319,71 1,68256435643564E-19
GO:0010629 negative regulation of gene expression 1472 454 241,84 1,46184086021505E-20
GO:0061919 process utilizing autophagic mechanism 392 120 64,4 3,65003846153846E-09
GO:0009894 regulation of catabolic process 732 221 120,26 0,000681573529412
GO:0008652 cellular amino acid biosynthetic process 83 25 13,64 4,96575E-05
GO:0010628 positive regulation of gene expression 1621 488 266,32 5,05603636363636E-18
GO:1905114 cell surface receptor signaling pathway ... 482 145 79,19 7,25431304347826E-17
GO:0048545 response to steroid hormone 334 100 54,87 1,72614525139665E-09
GO:0031324 negative regulation of cellular metaboli... 2065 618 339,27 1,69103918918919E-25
GO:0006355 regulation of transcription, DNA-templat... 2729 813 448,36 2,97812048192771E-22
GO:2001141 regulation of RNA biosynthetic process 2778 827 456,41 3,11471774193548E-15
GO:0006351 transcription, DNA-templated 2873 853 472,01 2,37676923076923E-28
GO:0007034 vacuolar transport 118 35 19,39 1,99837810945274E-08
GO:0032774 RNA biosynthetic process 2917 862 479,24 2,37676923076923E-28
GO:0007389 pattern specification process 349 103 57,34 5,70727388535032E-11
GO:0019219 regulation of nucleobase-containing comp... 3217 943 528,53 2,95093258426966E-21
GO:0071407 cellular response to organic cyclic comp... 502 147 82,48 9,77784810126582E-11
GO:0060249 anatomical structure homeostasis 352 103 57,83 8,54091056910569E-16
GO:0034654 nucleobase-containing compound biosynthe... 3345 973 549,56 9,32769811320755E-19
GO:0018130 heterocycle biosynthetic process 3399 985 558,43 2,37676923076923E-28
GO:0010605 negative regulation of macromolecule met... 2154 624 353,89 2,40317777777778E-09
GO:0090304 nucleic acid metabolic process 4138 1194 679,84 2,37676923076923E-28
GO:0019438 aromatic compound biosynthetic process 3406 981 559,58 2,37676923076923E-28
GO:0048511 rhythmic process 247 71 40,58 4,00320802919708E-05
GO:2000112 regulation of cellular macromolecule bio... 3101 890 509,47 1,80934234234234E-17
GO:0010556 regulation of macromolecule biosynthetic... 3207 913 526,89 1,70335128205128E-23
GO:0051252 regulation of RNA metabolic process 2978 844 489,26 9,90751086956522E-21
GO:1901362 organic cyclic compound biosynthetic pro... 3535 992 580,78 2,37676923076923E-28
GO:0044248 cellular catabolic process 1935 541 317,91 1,14823648648649E-11
GO:0061458 reproductive system development 343 95 56,35 4,26370250896057E-05
GO:0022412 cellular process involved in reproductio... 261 72 42,88 0,000929595988539
GO:0048608 reproductive structure development 341 94 56,02 0,000139041
7 GO:0009893 positive regulation of metabolic process 2921 805 479,9 2,37676923076923E-28
GO:1901575 organic substance catabolic process 1828 503 300,33 5,24869871794872E-11
GO:0033157 regulation of intracellular protein tran... 200 55 32,86 1,52314084507042E-07
GO:0031326 regulation of cellular biosynthetic proc... 3338 917 548,41 1,00754347826087E-26
GO:0009892 negative regulation of metabolic process 2327 639 382,31 2,25297916666667E-20
GO:0009889 regulation of biosynthetic process 3399 925 558,43 2,37676923076923E-28
GO:0006139 nucleobase-containing compound metabolic... 4630 1251 760,68 2,37676923076923E-28
GO:0030099 myeloid cell differentiation 336 90 55,2 0,00030898
GO:0007017 microtubule-based process 602 161 98,9 1,58082790697674E-07
GO:0000003 reproduction 1101 292 180,89 6,15225663716814E-17
GO:0022414 reproductive process 1098 291 180,39 7,85768103448276E-17
GO:0046483 heterocycle metabolic process 4776 1264 784,66 2,37676923076923E-28
GO:0003006 developmental process involved in reprod... 518 137 85,1 0,000279553333333
GO:0016197 endosomal transport 174 46 28,59 9,60559585492228E-09
GO:0044260 cellular macromolecule metabolic process 6674 1763 1096,49 2,37676923076923E-28
GO:0044419 interspecies interaction between organis... 737 194 121,08 4,0553625E-06
GO:0006725 cellular aromatic compound metabolic pro... 4819 1264 791,73 1,50076E-26
GO:0009056 catabolic process 2174 564 357,17 3,88335519125683E-09
GO:0009790 embryo development 772 200 126,83 5,14966666666667E-08
GO:0051171 regulation of nitrogen compound metaboli... 4663 1205 766,1 6,621E-15
GO:0009059 macromolecule biosynthetic process 4059 1048 666,87 4,56024698795181E-10
GO:0009628 response to abiotic stimulus 951 245 156,24 6,11350802139037E-09
GO:0044271 cellular nitrogen compound biosynthetic ... 3956 1012 649,94 2,57483333333333E-18
GO:1903706 regulation of hemopoiesis 342 87 56,19 0,000296348742138
GO:1901360 organic cyclic compound metabolic proces... 5009 1272 822,94 8,44545333333333E-25
GO:0071396 cellular response to lipid 511 129 83,95 7,58898245614035E-05
GO:0048523 negative regulation of cellular process 3779 947 620,86 2,37676923076923E-28
GO:0060255 regulation of macromolecule metabolic pr... 4858 1217 798,14 0,000239210322581
GO:0097190 apoptotic signaling pathway 496 124 81,49 9,23247011952191E-06
GO:0010468 regulation of gene expression 3522 877 578,64 1,33181034482759E-06
GO:0031323 regulation of cellular metabolic process 4883 1215 802,24 2,37676923076923E-28
GO:0051338 regulation of transferase activity 803 199 131,93 8,13105263157895E-09
GO:0034641 cellular nitrogen compound metabolic pro... 5227 1294 858,76 6,68466346153846E-19
GO:0002520 immune system development 756 185 124,21 2,00264814814815E-10
GO:0006950 response to stress 3200 770 525,74 2,37676923076923E-28
GO:0043170 macromolecule metabolic process 7510 1803 1233,84 2,37676923076923E-28
GO:0048519 negative regulation of biological proces... 4189 987 688,22 2,37676923076923E-28
GO:0040007 growth 764 179 125,52 1,04219444444444E-05
GO:0016043 cellular component organization 5129 1190 842,66 2,37676923076923E-28
GO:0006807 nitrogen compound metabolic process 8145 1877 1338,17 2,37676923076923E-28
GO:1903827 regulation of cellular protein localizat... 435 100 71,47 2,93335443037975E-06
GO:0032504 multicellular organism reproduction 625 141 102,68 7,09671378091873E-05
GO:0033365 protein localization to organelle 781 175 128,31 4,03348275862069E-08
GO:0007178 transmembrane receptor protein serine/th... 282 63 46,33 9,08764705882353E-05
GO:0044238 primary metabolic process 8586 1879 1410,62 4,99820588235294E-19
GO:0044237 cellular metabolic process 8695 1900 1428,53 2,37676923076923E-28
GO:0009968 negative regulation of signal transducti... 1022 218 167,91 3,96128205128205E-05
7 GO:0071704 organic substance metabolic process 8885 1895 1459,74 9,57017699115044E-07
GO:0008152 metabolic process 9233 1929 1516,92 2,37676923076923E-28
GO:0051704 multi-organism process 1984 397 325,96 0,000147861716172
GO:0032386 regulation of intracellular transport 324 61 53,23 0,000643708333333
GO:0009991 response to extracellular stimulus 433 80 71,14 0,000120449830508
GO:0009987 cellular process 12542 2175 2060,56 2,37676923076923E-28
GO:0046390 ribose phosphate biosynthetic process 226 38 37,13 0,000514966666667
GO:0006796 phosphate-containing compound metabolic ... 2686 450 441,29 2,2237196969697E-05
GO:0070727 cellular macromolecule localization 1542 257 253,34 0,000603114114114
GO:0098661 inorganic anion transmembrane transport 91 15 14,95 4,1421231884058E-05
GO:0002218 activation of innate immune response 206 33 33,84 0,000429138888889
GO:0008104 protein localization 2255 341 370,48 0,000767932748538
GO:0071705 nitrogen compound transport 1944 223 319,39 0,0009655625
GO:0043101 purine-containing compound salvage 15 7 0,26 0,000180238333333
GO:0046033 AMP metabolic process 13 6 0,23 0,000188821111111
GO:0016180 snRNA processing 24 11 0,42 9,40373913043478E-05
GO:0042795 snRNA transcription by RNA polymerase II 66 25 1,16 5,40715E-20
GO:0098781 ncRNA transcription 90 30 1,58 4,78919E-23
GO:0031163 metallo-sulfur cluster assembly 22 7 0,39 1,17706666666667E-05
GO:0016073 snRNA metabolic process 83 25 1,45 2,05251E-09
GO:0043094 cellular metabolic compound salvage 34 10 0,6 1,85388E-06
GO:0009163 nucleoside biosynthetic process 39 11 0,68 9,75726315789474E-06
GO:1901659 glycosyl compound biosynthetic process 42 11 0,74 0,000346270689655
GO:0006383 transcription by RNA polymerase III 45 11 0,79 1,17706666666667E-05
GO:0009116 nucleoside metabolic process 109 24 1,91 7,7245E-16
GO:1901657 glycosyl compound metabolic process 130 24 2,28 6,43708333333333E-11
8 GO:0072527 pyrimidine-containing compound metabolic... 80 14 1,4 2,22079375E-06
GO:1901293 nucleoside phosphate biosynthetic proces... 284 30 4,97 7,27011764705882E-06
GO:0006457 protein folding 191 20 3,34 2,13909230769231E-07
GO:0006753 nucleoside phosphate metabolic process 523 45 9,16 5,020925E-15
GO:0055086 nucleobase-containing small molecule met... 597 48 10,45 2,10668181818182E-08
GO:0034660 ncRNA metabolic process 464 36 8,12 5,18645E-07
GO:0072521 purine-containing compound metabolic pro... 432 28 7,56 0,0002041475
GO:0019637 organophosphate metabolic process 965 45 16,89 0,000188821111111
GO:0018130 heterocycle biosynthetic process 3399 120 59,5 0,000381075333333
GO:1901362 organic cyclic compound biosynthetic pro... 3535 123 61,88 0,000185388
8 GO:0044237 cellular metabolic process 8695 192 152,2 7,81032777777778E-06
GO:0019081 viral translation 13 13 1,12 2,23152222222222E-09
GO:0010603 regulation of cytoplasmic mRNA processin... 7 7 0,6 7,13030769230769E-07
GO:0075522 IRES-dependent viral translational initi... 10 10 0,86 7,20953333333333E-07
GO:0010606 positive regulation of cytoplasmic mRNA ... 5 5 0,43 0,000313457971014
GO:0006418 tRNA aminoacylation for protein translat... 44 41 3,79 5,25923404255319E-09
GO:0043039 tRNA aminoacylation 47 43 4,04 1,47044698795181E-11
GO:0043038 amino acid activation 48 43 4,13 2,86092592592593E-28
GO:0044417 translocation of molecules into host 8 7 0,69 7,06582170542636E-05
GO:0006406 mRNA export from nucleus 102 87 8,78 2,86092592592593E-28
GO:0006369 termination of RNA polymerase II transcr... 33 28 2,84 6,65905172413793E-26
GO:0071166 ribonucleoprotein complex localization 122 102 10,5 2,86092592592593E-28
GO:0071426 ribonucleoprotein complex export from nu... 121 101 10,41 2,86092592592593E-28
GO:0033962 cytoplasmic mRNA processing body assembl... 17 14 1,46 2,2793606557377E-05
GO:0006405 RNA export from nucleus 128 105 11,01 2,6110985915493E-16
GO:0033119 negative regulation of RNA splicing 21 17 1,81 6,6382421875E-05
GO:0006364 rRNA processing 174 138 14,97 7,51572972972973E-16
GO:0000375 RNA splicing, via transesterification re... 303 240 26,07 2,09205208333333E-08
GO:0000398 mRNA splicing, via spliceosome 300 237 25,81 5,86670886075949E-13
GO:1905216 positive regulation of RNA binding 9 7 0,77 7,24914615384615E-05
GO:0000956 nuclear-transcribed mRNA catabolic proce... 190 145 16,35 9,43202105263158E-09
GO:0006613 cotranslational protein targeting to mem... 98 74 8,43 7,84333846153846E-21
GO:0002181 cytoplasmic translation 88 66 7,57 2,55121100917431E-06
GO:0022613 ribonucleoprotein complex biogenesis 394 291 33,9 2,86092592592593E-28
GO:0033750 ribosome localization 15 11 1,29 4,23822123893805E-06
GO:0045047 protein targeting to ER 104 76 8,95 2,86092592592593E-28
GO:0016072 rRNA metabolic process 201 144 17,29 1,09273414634146E-11
GO:0008380 RNA splicing 378 270 32,52 3,65460215053763E-09
GO:0000921 septin ring assembly 14 10 1,2 0,000232315789474
GO:0022618 ribonucleoprotein complex assembly 213 152 18,33 2,86092592592593E-28
GO:0006413 translational initiation 185 131 15,92 2,86092592592593E-28
GO:0072599 establishment of protein localization to... 108 76 9,29 1,710425E-27
GO:0042255 ribosome assembly 54 38 4,65 7,04954368932039E-07
GO:0050657 nucleic acid transport 174 122 14,97 3,19055434782609E-09
GO:0071826 ribonucleoprotein complex subunit organi... 224 156 19,27 2,86092592592593E-28
C GO:0051236 establishment of RNA localization 177 122 15,23 2,86092592592593E-28
GO:0006397 mRNA processing 439 293 37,77 2,2793606557377E-05
GO:0032185 septin cytoskeleton organization 15 10 1,29 2,08301123595506E-09
GO:1905214 regulation of RNA binding 12 8 1,03 9,42650847457627E-06
GO:0006611 protein export from nucleus 166 108 14,28 2,86092592592593E-28
GO:0006403 RNA localization 208 135 17,9 2,86092592592593E-28
GO:0006402 mRNA catabolic process 274 174 23,58 2,86092592592593E-28
GO:0051168 nuclear export 178 113 15,32 6,58261739130435E-06
GO:0050779 RNA destabilization 19 12 1,63 0,000313457971014
GO:0016071 mRNA metabolic process 692 434 59,54 2,86092592592593E-28
GO:0015931 nucleobase-containing compound transport 201 123 17,29 2,86092592592593E-28
GO:0006450 regulation of translational fidelity 15 9 1,29 6,20989215686275E-07
GO:0006401 RNA catabolic process 304 181 26,16 2,86092592592593E-28
GO:0006396 RNA processing 781 464 67,2 2,86092592592593E-28
GO:0070972 protein localization to endoplasmic reti... 128 76 11,01 6,80803389830509E-25
GO:0050684 regulation of mRNA processing 117 69 10,07 2,45222222222222E-22
GO:0032239 regulation of nucleobase-containing comp... 14 8 1,2 0,00092694
GO:0043484 regulation of RNA splicing 116 65 9,98 1,75242388059702E-19
GO:0090501 RNA phosphodiester bond hydrolysis 127 71 10,93 9,9315E-18
GO:0006353 DNA-templated transcription, termination 68 38 5,85 9,897015625E-22
GO:0043046 DNA methylation involved in gamete gener... 18 10 1,55 0,00024211119403
GO:1902369 negative regulation of RNA catabolic pro... 43 23 3,7 1,42805882352941E-05
GO:1903313 positive regulation of mRNA metabolic pr... 63 33 5,42 0,000344546043165
GO:1903311 regulation of mRNA metabolic process 227 118 19,53 2,86092592592593E-28
GO:0034660 ncRNA metabolic process 464 241 39,92 2,86092592592593E-28
GO:0043487 regulation of RNA stability 105 54 9,03 2,03276315789474E-26
GO:1902373 negative regulation of mRNA catabolic pr... 36 18 3,1 0,00039726
GO:0051169 nuclear transport 323 160 27,79 2,86092592592593E-28
GO:0001510 RNA methylation 61 30 5,25 3,61307258064516E-05
GO:0006412 translation 626 298 53,86 2,86092592592593E-28
GO:0061013 regulation of mRNA catabolic process 119 56 10,24 7,38865217391304E-18
GO:0043043 peptide biosynthetic process 644 298 55,41 2,86092592592593E-28
GO:0006612 protein targeting to membrane 166 76 14,28 2,57483333333333E-20
GO:0031503 protein-containing complex localization 239 102 20,56 2,86092592592593E-28
GO:0045727 positive regulation of translation 105 44 9,03 1,59879186046512E-10
GO:0043604 amide biosynthetic process 713 298 61,35 2,86092592592593E-28
GO:0016226 iron-sulfur cluster assembly 22 9 1,89 3,89704504504505E-06
GO:0010608 posttranscriptional regulation of gene e... 422 171 36,31 2,86092592592593E-28
GO:0006518 peptide metabolic process 763 305 65,65 2,86092592592593E-28
GO:0006417 regulation of translation 341 136 29,34 2,86092592592593E-28
GO:0034655 nucleobase-containing compound catabolic... 479 190 41,21 2,86092592592593E-28
GO:0017148 negative regulation of translation 173 65 14,89 4,48021E-07
GO:0046700 heterocycle catabolic process 526 192 45,26 2,86092592592593E-28
GO:0044270 cellular nitrogen compound catabolic pro... 526 192 45,26 2,86092592592593E-28
GO:0034248 regulation of cellular amide metabolic p... 381 139 32,78 2,86092592592593E-28
GO:0090305 nucleic acid phosphodiester bond hydroly... 241 87 20,74 2,71630769230769E-09
GO:0034250 positive regulation of cellular amide me... 125 45 10,76 1,63368735632184E-09
GO:0019439 aromatic compound catabolic process 540 192 46,46 2,86092592592593E-28
GO:0034249 negative regulation of cellular amide me... 189 66 16,26 2,08301123595506E-09
GO:0009451 RNA modification 128 44 11,01 1,08302268041237E-07
GO:0043603 cellular amide metabolic process 902 307 77,61 2,86092592592593E-28
GO:1901361 organic cyclic compound catabolic proces... 570 192 49,04 2,86092592592593E-28
GO:0006414 translational elongation 128 40 11,01 1,5449E-05
GO:0001825 blastocyst formation 23 7 1,98 0,000306708088235
GO:0016458 gene silencing 186 51 16 9,53641975308642E-12
GO:0018208 peptidyl-proline modification 48 13 4,13 7,45813793103448E-06
GO:0032204 regulation of telomere maintenance 73 18 6,28 2,14569444444444E-06
GO:0072395 signal transduction involved in cell cyc... 69 17 5,94 3,27706060606061E-07
GO:0071158 positive regulation of cell cycle arrest 75 18 6,45 0,000232315789474
GO:0006997 nucleus organization 110 26 9,46 3,01443902439024E-05
GO:0006886 intracellular protein transport 958 225 82,43 2,86092592592593E-28
GO:0043414 macromolecule methylation 249 58 21,42 5,04769306930693E-07
GO:0006998 nuclear envelope organization 49 11 4,22 0,00011793129771
GO:0032259 methylation 294 61 25,3 4,78183333333333E-05
GO:0034622 cellular protein-containing complex asse... 939 192 80,79 3,00397222222222E-16
GO:0010467 gene expression 4333 876 372,81 2,86092592592593E-28
GO:0044265 cellular macromolecule catabolic process 992 200 85,35 2,394595E-24
GO:0016070 RNA metabolic process 3693 738 317,75 2,40763636363636E-14
GO:1901566 organonitrogen compound biosynthetic pro... 1636 321 140,76 2,86092592592593E-28
C GO:0071156 regulation of cell cycle arrest 98 19 8,43 0,000438269503546
GO:0072331 signal transduction by p53 class mediato... 200 38 17,21 2,26585333333333E-15
GO:0090304 nucleic acid metabolic process 4138 763 356,04 2,86092592592593E-28
GO:0042770 signal transduction in response to DNA d... 123 22 10,58 2,14569444444444E-06
GO:0035722 interleukin-12-mediated signaling pathwa... 45 8 3,87 0,000297536296296
GO:0070727 cellular macromolecule localization 1542 267 132,67 2,86092592592593E-28
GO:0009057 macromolecule catabolic process 1189 204 102,3 4,0001875E-06
GO:0034613 cellular protein localization 1533 261 131,9 7,04292647058824E-19
GO:0046907 intracellular transport 1531 259 131,73 2,86092592592593E-28
GO:1901796 regulation of signal transduction by p53... 120 19 10,32 6,56886614173228E-05
GO:0044085 cellular component biogenesis 2670 397 229,73 5,33692727272727E-28
GO:0044419 interspecies interaction between organis... 737 107 63,41 2,66846363636364E-06
GO:0009892 negative regulation of metabolic process 2327 336 200,22 1,52672470588235E-10
GO:0033044 regulation of chromosome organization 307 44 26,41 3,70776E-05
GO:0045184 establishment of protein localization 1760 252 151,43 2,86092592592593E-28
GO:0051649 establishment of localization in cell 1838 263 158,14 2,86092592592593E-28
GO:0065003 protein-containing complex assembly 1601 229 137,75 3,476025E-12
GO:0043933 protein-containing complex subunit organ... 1878 260 161,58 2,00077213114754E-23
GO:0034645 cellular macromolecule biosynthetic proc... 3932 534 338,31 2,86092592592593E-28
GO:0071705 nitrogen compound transport 1944 261 167,26 2,86092592592593E-28
GO:0009059 macromolecule biosynthetic process 4059 542 349,24 0,000702227272727
GO:0043170 macromolecule metabolic process 7510 965 646,17 2,86092592592593E-28
GO:0071702 organic substance transport 2254 265 193,94 9,21956451612903E-23
GO:0006807 nitrogen compound metabolic process 8145 950 700,8 1,12189166666667E-10
GO:0071704 organic substance metabolic process 8885 990 764,47 0,000708079166667
GO:0044237 cellular metabolic process 8695 967 748,12 2,86092592592593E-28
GO:0032504 multicellular organism reproduction 625 61 53,78 7,5264358974359E-06
C GO:0033554 cellular response to stress 1625 117 139,82 9,91067924528302E-07
GO:0010033 response to organic substance 2704 145 232,65 0,000576617605634
GO:0030150 protein import into mitochondrial matrix 17 14 0,6 0,000809233333333
GO:0015986 ATP synthesis coupled proton transport 22 18 0,77 2,97536296296296E-09
GO:0042776 mitochondrial ATP synthesis coupled prot... 17 13 0,6 4,5814275862069E-09
GO:0044743 protein transmembrane import into intrac... 38 29 1,34 2,5104625E-24
GO:0018206 peptidyl-methionine modification 11 8 0,39 3,00629189189189E-08
GO:0007007 inner mitochondrial membrane organizatio... 38 27 1,34 5,98025806451613E-09
GO:0015919 peroxisomal membrane transport 17 12 0,6 3,51113636363636E-06
GO:0017004 cytochrome complex assembly 25 15 0,88 1,9863E-11
GO:0006474 N-terminal protein amino acid acetylatio... 15 9 0,53 1,3148085106383E-05
GO:0070525 tRNA threonylcarbamoyladenosine metaboli... 10 6 0,35 2,67138958333333E-05
GO:0065002 intracellular protein transmembrane tran... 53 29 1,86 1,33423181818182E-21
GO:0071806 protein transmembrane transport 60 31 2,11 1,67364166666667E-19
GO:0031365 N-terminal protein amino acid modificati... 24 12 0,84 1,69939E-11
GO:0006119 oxidative phosphorylation 113 56 3,97 2,207E-27
GO:1990542 mitochondrial transmembrane transport 75 36 2,64 1,64145625E-15
GO:0099131 ATP hydrolysis coupled ion transmembrane... 31 14 1,09 8,29668518518519E-05
GO:0022904 respiratory electron transport chain 97 43 3,41 1,3148085106383E-05
GO:0099132 ATP hydrolysis coupled cation transmembr... 52 23 1,83 1,67364166666667E-10
GO:1902600 proton transmembrane transport 134 59 4,71 2,207E-27
GO:0090662 ATP hydrolysis coupled transmembrane tra... 32 14 1,13 0,00015173125
GO:0006626 protein targeting to mitochondrion 89 32 3,13 6,83321153846154E-05
GO:0033108 mitochondrial respiratory chain complex ... 80 28 2,81 5,98025806451613E-09
GO:0010257 NADH dehydrogenase complex assembly 56 19 1,97 1,66373846153846E-09
GO:0022900 electron transport chain 166 55 5,84 2,37676923076923E-19
GO:0033572 transferrin transport 35 11 1,23 0,000314216949153
GO:0046034 ATP metabolic process 220 68 7,73 2,207E-27
GO:0045333 cellular respiration 153 46 5,38 1,18838461538462E-07
GO:0007006 mitochondrial membrane organization 111 33 3,9 2,09665E-06
GO:0070585 protein localization to mitochondrion 126 32 4,43 3,44091363636364E-11
GO:0006839 mitochondrial transport 205 52 7,21 4,17123E-22
11 GO:0009141 nucleoside triphosphate metabolic proces... 273 68 9,6 4,14683684210526E-13
GO:0009123 nucleoside monophosphate metabolic proce... 282 69 9,91 2,91814444444444E-13
GO:0007031 peroxisome organization 78 17 2,74 5,04457142857143E-05
GO:0007005 mitochondrion organization 437 95 15,36 2,207E-27
GO:0006415 translational termination 96 20 3,38 3,0898E-06
GO:0015980 energy derivation by oxidation of organi... 232 46 8,16 6,1796E-06
GO:0032543 mitochondrial translation 121 22 4,25 6,48858E-05
GO:0000041 transition metal ion transport 100 18 3,52 1,84634390243902E-06
GO:0006091 generation of precursor metabolites and ... 412 74 14,49 2,207E-27
GO:0017038 protein import 192 34 6,75 2,40317777777778E-08
GO:0140053 mitochondrial gene expression 139 24 4,89 1,18139411764706E-08
GO:0006163 purine nucleotide metabolic process 401 68 14,1 0,000411973333333
GO:0019693 ribose phosphate metabolic process 413 68 14,52 2,207E-27
GO:0072521 purine-containing compound metabolic pro... 432 68 15,19 4,11973333333333E-24
GO:0006414 translational elongation 128 20 4,5 7,31794736842105E-08
GO:0015672 monovalent inorganic cation transport 439 64 15,43 4,30365E-09
GO:0006605 protein targeting 371 47 13,04 1,85388E-08
GO:0055086 nucleobase-containing small molecule met... 597 73 20,99 2,8691E-19
GO:0017144 drug metabolic process 694 72 24,4 2,45366470588235E-13
GO:0055085 transmembrane transport 1229 124 43,21 4,32572E-19
GO:0008286 insulin receptor signaling pathway 119 12 4,18 0,000506524590164
GO:0072594 establishment of protein localization to... 500 46 17,58 0,000200565964912
GO:0055114 oxidation-reduction process 863 75 30,34 1,14188260869565E-10
GO:0050954 sensory perception of mechanical stimulu... 126 10 4,43 0,000120783090909
GO:0019637 organophosphate metabolic process 965 76 33,93 5,994212E-10
GO:0033365 protein localization to organelle 781 54 27,46 1,4290325E-06
GO:0044281 small molecule metabolic process 1779 100 62,55 0,000237062241379
GO:0006996 organelle organization 3158 159 111,03 0,000622943548387
GO:0051234 establishment of localization 4186 200 147,17 7,2872641509434E-05
GO:1901663 quinone biosynthetic process 13 10 0,73 1,606696E-10
GO:0006743 ubiquinone metabolic process 14 10 0,79 1,34627E-09
D GO:0010257 NADH dehydrogenase complex assembly 56 27 3,15 5,14966666666667E-16
GO:0035510 DNA dealkylation 30 14 1,69 0,000820728125
GO:0032196 transposition 20 9 1,13 0,000241810434783
GO:0033108 mitochondrial respiratory chain complex ... 80 32 4,5 1,23592E-16
GO:1901661 quinone metabolic process 26 10 1,46 2,40317777777778E-07
GO:0042454 ribonucleoside catabolic process 21 8 1,18 0,000908764705882
GO:0033572 transferrin transport 35 13 1,97 1,0428075E-07
GO:0018195 peptidyl-arginine modification 25 9 1,41 4,6347E-05
GO:0072512 trivalent inorganic cation transport 38 13 2,14 4,91559090909091E-06
GO:0099131 ATP hydrolysis coupled ion transmembrane... 31 10 1,75 0,000452815517241
GO:0090662 ATP hydrolysis coupled transmembrane tra... 32 10 1,8 0,000514966666667
GO:0022904 respiratory electron transport chain 97 30 5,46 4,89218333333333E-05
GO:0006119 oxidative phosphorylation 113 32 6,36 1,23592E-16
GO:0009451 RNA modification 128 35 7,21 1,6607675E-11
GO:0099132 ATP hydrolysis coupled cation transmembr... 52 12 2,93 0,00092694
GO:0003341 cilium movement 41 9 2,31 0,000429138888889
GO:0008033 tRNA processing 106 23 5,97 5,87062E-05
GO:0022900 electron transport chain 166 31 9,35 2,75875E-05
D GO:0000041 transition metal ion transport 100 17 5,63 6,47386666666667E-05
GO:1902600 proton transmembrane transport 134 22 7,54 5,87062E-05
GO:0046034 ATP metabolic process 220 34 12,39 1,34627E-09
GO:0015980 energy derivation by oxidation of organi... 232 35 13,06 0,000415934615385
GO:0008286 insulin receptor signaling pathway 119 16 6,7 4,0553625E-05
GO:0007005 mitochondrion organization 437 51 24,6 4,0553625E-05
GO:0019693 ribose phosphate metabolic process 413 40 23,25 1,30722307692308E-05
GO:0046483 heterocycle metabolic process 4776 322 268,88 0,000797367741935
GO:0006725 cellular aromatic compound metabolic pro... 4819 324 271,3 0,000889487878788
GO:2000112 regulation of cellular macromolecule bio... 3101 206 174,58 0,00030898
GO:1901360 organic cyclic compound metabolic proces... 5009 331 281,99 0,000370776
GO:0010556 regulation of macromolecule biosynthetic... 3207 208 180,55 2,93531E-07
GO:0009889 regulation of biosynthetic process 3399 212 191,35 0,000987019444444
GO:0006996 organelle organization 3158 175 177,79 0,000430365
GO:0009607 response to biotic stimulus 790 34 7,5 4,78919E-14
GO:0043207 response to external biotic stimulus 765 34 7,26 1,5449E-07
GO:0051704 multi-organism process 1984 49 18,84 2,21435666666667E-07
GO:0098542 defense response to other organism 407 29 3,86 4,01674E-07
GO:0006952 defense response 1288 35 12,23 4,01674E-07
GO:0042742 defense response to bacterium 207 28 1,97 4,37721666666667E-07
E GO:0045087 innate immune response 710 32 6,74 1,67732E-06
GO:0009605 response to external stimulus 1929 37 18,31 1,40757555555556E-05
GO:0009620 response to fungus 41 11 0,39 0,000494368
GO:0071216 cellular response to biotic stimulus 190 11 1,8 0,000632004545455
GO:0009617 response to bacterium 504 34 4,79 0,000653611538462
GO:0051673 membrane disruption in other organism 10 6 0,09 0,000653611538462
Table fields:
Cluster - Cluster number on first time-interval

GO.ID - Gene Ontology term ID
Term - Human-readable term name
Annotated - Number of genes annotated for a specific GO term
Significant - Number of differentially expressed genes identified
Expected - Number of differentially expressed genes expected by the topGO statistical test
pValue - p-values adjusted by the Benjamini-Hochberg procedure.

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UNIVERSIDADE FEDERAL DO RIO GRANDE DO NORTE

INSTITUTO METRÓPOLE DIGITAL

Clovis Ferreira dos Reis

Análise Baseada em Biologia de Sistemas de

Clovis Ferreira dos Reis

Análise Baseada em Biologia de Sistemas de Dados

Orientador: Prof. Dr. Rodrigo Juliani Siqueira Dalmolin – UFRN

Análise Baseada em Biologia de Sistemas de Dados

Tese apresentada ao Programa de Pós-graduação

Área de concentração: Bioinformática

Linha de Pesquisa: Genômica

Orientador: Prof. Dr. Rodrigo Juliani S. Dalmolin

Reis, Clovis Ferreira dos.

Tese (Doutorado)-Universidade Federal do Rio Grande do Norte,

1. Exposição ao chumbo - Tese. 2. Análise de transcriptoma -

RN/UF/BCZM CDU 004:577

Elaborado por Raimundo Muniz de Oliveira - CRB-15/429

Aos colegas do BioME pela convivência;

1 CONSIDERAÇÕES SOBRE O CHUMBO . . . . . . . . . . . . . . . 13

APÊNDICE A – MATERIAL SUPLEMENTAR DO ARTIGO . . . . 61

Figura 1 – Número de casos de intoxicação por chumbo em crianças (USA) . . . . 15

Tabela 1 – Exemplo de enriquecimento de ontologias. . . . . . . . . . . . . . . . . 29

A.C. – Antes de Cristo

1 Considerações sobre o chumbo

– Maleabilidade, podendo ser facilmente reduzido a lâminas finas;

– Ductibilidade, podendo deformar-se sem romper-se sob pressão cisalhante;

– Baixa condutividade elétrica;

– Baixo ponto de fusão, tornando-se lı́quido a 327,4o C e;

– Altamente resistente à corrosão.

1.1 O chumbo como uma questão de saúde pública

contaminação ambiental. A gestão de resı́duos contaminados e a proibição do uso de aditivos à

• Poeira de tinta de casas antigas; • Fundições;

• Água proveniente de encanamentos anti-

• Rejeitos de mineradoras; • Alimentos contaminados; etc.

Alguns grupos populacionais também são especialmente suscetı́veis à contaminação por

Figura 1 – Número de casos confirmados de intoxicação por chumbo em crianças com

1.2 Efeitos neurotóxicos do chumbo

1.2.1 Causas da severa toxicidade do chumbo

Desempenha papel fundamental na estabilização da estrutura terciária de uma famı́lia de

1.3 O impacto sistêmico do envenenamento por chumbo

• Diminuir a frequência de abertura de canais associados a receptores de acetilcolina;

• Interferir em canais de cálcio dependentes de voltagem, impedindo o seu correto fun-

• Substituir ı́ons Ca2+ em metaloproteı́nas e receptores ionotrópicos, causando sua perda

• Substitui o Zn2+ em receptores NMDAR em vias glutamatérgicas, conhecidos por serem

• Causar estresse oxidativo, etc. [23]

Despolarização de Membrana Neurotransmissores e hormônios

Canais de Cálcio Adenilato Ciclase

O Glutationa Proteina Quinase C

O processo de diferenciação celular ocorre em diversas fases sucessivas, onde células

Figura 4 – Modelo esquemático da diferenciação neuronal a partir de células-tronco

2.1 Caracterizando a diferenciação neuronal em um transcriptoma

2.1.1 Genes marcadores de NPCs

e astrócitos, mas nunca em neurônios ou oligodendrócitos. Podem ainda ser encontrados

Inicialmente identificada em células tronco neuroepiteliais de ratos, a Nestina é um indicador

Sox1 é um membro da famı́lia de fatores de transcrição SoxB1 e sua expressão está

2.1.2 Genes marcadores neuronais

A TH é uma enzima considerada crı́tica para o funcionamento de diversos tipos de neurônios,

Membro da subfamı́lia NEUROD, pertencente ao grupo dos fatores de transcrição basic-

Seus elevados nı́veis de expressão estão associados ao desenvolvimento de neurônios maduros a

A doublecortina (DCX) é uma fosfoproteı́na necessária ao desenvolvimento dos microtúbulos.

3.1 O pacote transcriptogramer

3.1.1 Sequencia ordenada de genes

(a) Ordem aleatória (b) Troca aleatória de posição (c) Ordenado