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Troy Quintana
Biology 1615-018
Jack Later
Friday, April 11, 2014

















http://gosc.pl/files/11/12/16/058161_gn47s52a_34.jpg
Trisomy 21-derived miRNAs
provide an etiological basis for
aberrant protein expression in
human Down Syndrome Brains

T.S ELTON, D.S. FELDMAN, E. HEAD, W.D. BECK, A.P PLEISTER, S.E.
SANSOM, G.E. MALANA, M.M MARTIN, A.V. TERRY JR, G.J NUOVO, AND
D.E. KUHN.
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1) Introduction
i) Failure of the 21
st
chromosome
ii) Down Syndrome (DS) or Trisomy 21 blamed for cognitive impairment and
congenital heart defects
iii) Target important micro RNA (miRNAs) that may play a role in DS phenotype.
2) Methods and Materials
i) Brain tissues derived from individuals with DS
ii) Cell culture-human neuroblastoma cell line and fetal bovine serum
iii) Human Chromosome 21 (Hsa21)-derived miRNA Bioinformatic analyses
iv) Real time PCR
v) Luciferase Reporter Constructs
vi) Transfection and Luciferase assay
vii) Western blot analyses
viii) Immunohistochemistry
ix) Chromatin Immunoprecipitation
x) Transgenic mice
xi) Stereotaxic Intracerebroventricular Injection
xii) Statistical analysis
3) Results
i) Hsa21 derived miRNAs are Overexpressed in Human DS brain specimens
ii) methyl-CpG-binding protein (MeCP-2) is a target of Hsa21-derived MiRNAs
iii) MeCP-2 is Under expressed in Human DS brain specimens
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iv) MeCP-2 target genes are aberrantly expressed in human DS brain specimens
v) Ts65Dn mouse model of DS aberrantly expresses miR-155/802, MeCP-2, and MeCP-
2 target genes.
4) Discussion
i) Improper attenuation of MeCP-2, CREB1, and MEF2C aberrantly expressed that
contributes to cognitive defects
ii) In vivo silencing of target miRNAs results in normalization of the mice.
iii) Transgenic mice demonstrate both under and over expression of MeCP-2 are
detrimental to cognitive development.
iv) MeCP-2 in relation to DS fetal fibroblasts and heart
v) Precise control of MeCP-2 is critical
vi) cAMP responsive element-binding protein (CREB) factors are critical in relation to
MeCP-2
Introduction:
This summary is extracted from one article, The Journal of Biological Chemistry (JBC),
Trisomy 21 or Down Syndrome (DS) is caused by a failure of the 21st chromosome to separate
during development. A sperm or egg cell is produced with an extra copy of chromosome 21; this
cell thus has 24 chromosomes instead of 23. When combined with a normal cell from the other
parent, the baby has 47 chromosomes instead of 46, with three copies of chromosome 21. The
incidence of one in 750 live births, DS is the most frequently survivable congenital chromosomal
abnormality. The phenotype includes cognitive impairment, congenital heart defects, craniofacial
abnormalities, gastrointestinal anomalies, leukemia, and Alzheimer disease. As stated in the JBC
article the aim of this study was to identify important Human chromosome 21 (Hsa21)-derived
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miRNA/mRNA target pairs that may play a role, in part, in mediating the DS phenotype. Past
studies have shown that (Hsa21) harbors five miRNAs genes; miR-99a, let-7c, miR-125b-2,
miR-155, and miR-802. According to this JBC article their lab recently demonstrated that Hsa21
derived miRNAs are over expressed in DS brain and heart specimens (Fitzpatrick D. R., 2002)
(LeJeune J., 1959).
Methods and Materials:
Various Human brain cerebellum, hippocampus, and pre-frontal cortex samples, were
obtained from the Brain and Tissue Bank for Developmental Disorders, University of Maryland
at Baltimore, in contract with National Institutes of health, NICHD. Fetal samples ranged from
18-22 weeks of gestation. Children, adolescent, and adult brain samples were obtained from
specimens ranging 1-8, 9-19 and 20-50 years of age, respectively (Jiang J., 2008) (LeJeune J.,
1959).
Results:
Experiments demonstrated that all five Has21-derived miRNAs were over expressed in
human DS fetal heart and hippocampus specimens by at least 50% in prefrontal cortex samples
(Jiang J., 2008) (Fitzpatrick D. R., 2002). The Major finding was that the Hsa21-derived
miRNAS 155/-802 and proteins MeCP2, CREB1, and MEF2C are all aberrantly expressed in a
different manner than brain specimens isolated form DS individuals. Importantly was that in vivo
silencing (in which chemically modified, cholesterol conjugated, single-stranded RNA
complimentary to miRNAs, which were designatedantagomirs can silence endogenous
miRNAs in vivo) of miR-155/-802 resulted in normalization of the appropriate target proteins in
the transgenic mice.
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Conclusion:
The study suggests that the both under expression and over expression of MeCP2 are
detrimental to cognitive development which would indicate that level of MeCP2 in the central
nervous system are crucial for neural function and that precise control of MeCP2 is mandatory
for normal behavior. It also suggests that decreased MeCP2 may be indicative of
neurodevelopment disorders (Fitzpatrick D. R., 2002) (Jiang J., 2008) (LeJeune J., 1959).
Discussion:
The results that were attempted to be demonstrated here, that not only is it the extra
chromosome that causes cognitive defects and the physical attributes of the disorder, but also
may be contributed by other factors such as the micro RNAs, methyl-CpG- binding protein and
the CREB1/Creb1 and MEF2C/Mef2c genes. This study and studies like this can only improve
our understanding of the disorder and that if we can target the genes or binding proteins that
may or may not affect the disorder that may be the solution so that individuals can have a normal
life without having to live a life worrying about heart problems, appearance anomalies, and the
various other ailments that go along with Down Syndrome (Jiang J., 2008) (Fitzpatrick D. R.,
2002).





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Works Cited

Fitzpatrick D. R., R. J. (2002). Hum. Mol. Genet. PubMed, 3249-3256.
LeJeune J., G. <. (1959). Comptes Renus de l'Academic les Sciences. Journal Am. Med. Assoc.,
249.
Kuhn, Donald E., Gerard J. Nuovo, Alvin V. Terry, and T. S. Elton. "(2009)
21-derived MicroRNAs Provide an Etiological Basis for Aberrant Protein
Expression in Human Down Syndrome Brains." National Library of Medicine.
American Society for Biochemistry and Molecular Biology, 6 Nov. 2009. Web. 10
Apr. 2014.
<http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fpmc%2Farticles%2FPMC2801278
%2F>.