July 11, 2018

Learning Outcomes:
At the end of this chapter, the students should be able to:

1. describe gene therapy and its various forms;

2. discuss the prevalence of gene therapy in daily life;
3. explore the opportunities that may be opened by gene
therapy in the future; and
4. assess the issue’s potential benefits and detriments to
global health.
5. Compare and contrast gene therapy and stem cell therapy.
Genes
 Basic unit of heredity
 Carried on a chromosome
 Encode how to make a protein: DNA  RNA  proteins
Proteins carry out most of life’s function
 Causes dysfunction of a protein when altered
 When there is mutation in the gene, then it will:
Change the codon  change the amino acid  change the
conformation of a protein  change the function of a protein 
results to genetic disorders in the genome.
Gene Therapy seeks to alter genes to correct genetic
defects in order to prevent or cure genetic diseases.
How it started….
In the 1980s, Scientists began to look into gene therapy.
 They would insert human genes into a bacteria cell.
 Then the bacteria cell would transcribe and translate the
information into a protein
 Then they would introduce the protein into human cells
The First Case….
The first gene therapy was performed on September 14th,
1990
 Ashanti De Silva was treated for SCID (Sever combined
immunodeficiency)
 Doctors removed her white blood cells, inserted the missing
gene into the WBC, and then put them back into her blood
stream.
 This strengthened her immune system
 Only worked for a few months 
 Two Basic Types of Gene Therapy
1. Somatic gene therapy
o insertion of therapeutic DNA into body cells
o effects of the therapy are confined to the individual being
treated

2. Germline gene therapy

o modification of the genes inside germ cells (sperm or ova)
o alters the genome of future generations to come

https://www.news-medical.net/health/Gene-Therapy-Types.aspx
1. Ex vivo – transfer of genes in cultured cells and
will be reinjected to the body of the patient.

2. In vivo – introduction of therapeutic gene into the

vector injected directly to the body.
 How it works?

A closer look on Ex vivo gene

therapy
Step 1. Treatment begins with cells being removed
from an infected person. These cells are
then grown and multiplies in a laboratory.
Step 2. Therapeutic gene will be introduced into the
cultured cells using a virus called Vector. The
virus’s DNA is taken out and replaced with the
one that treats the patient.
Step 3. The genetically transformed cells are selected
and reinjected into the patient’s body.

These cells can either destroy the tumour or they produce the
necessary proteins and hormones for treatment.
A closer look on In vivo gene
therapy
In vivo gene therapy
 Involves direct delivery of the therapeutic gene into the target cells
of a particular tissue of a patient
 Potential tissue candidates include liver, muscle, skin, spleen, lung, brain
and blood cells.
 Can be carried out using viral or non-viral vectors
 Success depends on:
 efficiency of the uptake of the therapeutic gene by the target cells
 intracellular degradation of the gene and its uptake by nucleus
 the expression capability of the gene
 Vector refers to carrier particles or molecules
used to deliver genes. This includes:

1. Viral vectors
2. Non-Viral vectors

http://www.biologydiscussion.com/gene/therapy/gene-therapy-ex-vivo-and-in-vivo-gene-therapy-with-diagram/9969
Viruses
 Replicate by inserting their DNA into a host cell
 Gene therapy can use this to insert genes that encode for a
desired protein to create the desired trait
 Four different types
1. Retroviruses
 Created double stranded DNA copies from RNA genome
o The retrovirus goes through reverse transcription using reverse
transcriptase and RNA
o The double stranded viral genome integrates into the human
genome using integrase
 Integrase inserts the gene anywhere because it has no specific
site
 May cause insertional mutagenesis
 One gene disrupts another gene’s code (disrupted cell division causes
cancer from uncontrolled cell division)
o Vectors used are derived from the human immunodeficiency
virus (HIV) and are being evaluated for safety
2. Adenoviruses
 Are double stranded DNA genome that cause respiratory,
intestinal, and eye infections in humans
 The inserted DNA is not incorporate into genome
 Not replicated though 
 Has to be reinserted when more cells divide
 Ex. Common cold
3. Adeno-associated Viruses
Small, single stranded DNA that insert genetic material at a specific point on
chromosome 19
From parvovirus family- causes no known disease and doesn't trigger patient
immune response.
Low information capacity
gene is always "on" so the protein is always being expressed, possibly even in
instances when it isn't needed.
hemophilia treatments, for example, a gene-carrying vector could be injected
into a muscle, prompting the muscle cells to produce Factor IX and thus prevent
bleeding.
Study by Wilson and Kathy High (University of Pennsylvania), patients have
not needed Factor IX injections for more than a year
4. Herpes Simplex Viruses
• Double stranded DNA viruses that infect neurons
• Ex. Herpes simplex virus type 1

http://www.ucmp.berkeley.edu/alllife/virus.html
Non-viral Vectors
o Direct introduction of therapeutic DNA
 But only with certain tissue
 Requires a lot of DNA
o Creation of artificial lipid sphere with aqueous core, liposome
 Carries therapeutic DNA through membrane
o Chemically linking DNA to molecule that will bind to special cell receptors
 DNA is engulfed by cell membrane
 Less effective 
o Trying to introduce a 47th chromosome
 Exist alongside with the 46 others
 Could carry a lot of information
 But how to get the big molecule through membranes?
Current Status
 FDA hasn’t approved any human gene therapy product for sale

Reasons:
 In 1999, 18-year-old Jesse Gelsinger died from multiple organ failure
4 days after treatment for omithine transcarboxylase deficiency.
 Death was triggered by severe immune response to adenovirus carrier
 January 2003, halt to using retrovirus vectors in blood stem cells
because children developed leukemia-like condition after successful
treatment for X-linked severe combined immunodeficiency disease
Problems with Gene Therapy
Short Lived
o Hard to rapidly integrate therapeutic DNA into genome and rapidly dividing
nature of cells prevent gene therapy from long time
o Would have to have multiple rounds of therapy
 Immune Response
o new things introduced leads to immune response
o increased response when a repeat offender enters
Viral Vectors
o patient could have toxic, immune, inflammatory response
o also may cause disease once inside
Multigene Disorders
o Heart disease, high blood pressure, Alzheimer’s, arthritis and diabetes are
hard to treat because you need to introduce more than one gene
May induce a tumor if integrated in a tumor suppressor gene because insertional
mutagenesis
 Gene Therapy
vs
Stem Cell Therapy
Stem Cells
Importance of Stem Cell Research
 Stem Cell History
1998 - Researchers first extract stem cells from human
embryos
1999 - First Successful human transplant of insulin-making
cells from cadavers
2001 - President Bush restricts federal funding for embryonic
stem-cell research
2002 - Juvenile Diabetes Research Foundation International
creates $20 million fund-raising effort to support stem-
cell research
2002 - California supports stem cell research
2004 - Harvard researchers grow stem cells from embryos
using private funding
2004 - Ballot measure for $3 Billion bond for stem cells
 Stem Cell History
2005 – Woo Suk Hwang reports that his team has used
therapeutic cloning. But his claims turned out to be false
later that year.
2006 – Shinya Yamanaka reveals that her team found a way
of making embryonic-like cells from adult cells –
avoiding the need to destroy the embryo.
2007 – Evans shares the Nobel prize for medicine with Mario
Capecchi and Oliver Smithies for work on genetics and
embryonic stem cells.
2009 – President Barack Obama lifts 2001 restrictions on
federal funding for human embryonic stem cell research
2010 – A person with spinal injury becomes the first to receive
a medical treatment derived from human embryonic
stem cells
 Stem Cell History
2012 – Human embryonic stem cells show medical promise in a
treatment that eases blindness
– Yamanaka wins a Nobel prize for creating induced
pluripotent stem cells, which he shares with John Gurdon
2013 – Shoukhrat Mitalipov and his colleagues produce human
embryonic stem cells from fetal cells using therapeutic
cloning – the breakthrough falsely claimed in 2005.
2014 – Charles Vacanti together with Haruko Obokata and
colleagues announced a revolutionary discovery that any cell
can potentially be rewound to a pre-embryonic state – using
a simple, 30-minute technique.
– Teams led by Dieter Egli and Young Gie
Chung, independently produce human embryonic stem cells
from adult cells, using therapeutic cloning.
– Masayo Takahashi selects patients to be the world’s first trial
of a therapy based on induced pluripotent stem cells, to treat
a form of age-related blindness.
 Stem Cell

•A cell that has the ability to

continuously divide and differentiate
(develop) into various other kind(s)
of cells/tissues
 Stem Cell Characteristics

• ‘Blank cells’ (unspecialized)

• Capable of dividing and renewing themselves for long
periods of time (proliferation and renewal)
• Have the potential to give rise to specialized cell types
(differentiation)
Classification of Stem Cells
Stem cell
type Description Examples
Cells from early
Each cell can develop
Totipotent (1-3 days)
into a new individual
embryos
Some cells of
Cells can form any (over
Pluripotent blastocyst (5 to 14
200) cell types
days)
Cells differentiated, but Fetal tissue, cord
Multipotent can form a number of blood, and adult
other tissues stem cells
 This cell
Can form the
Embryo and placenta

This cell
Can just form the
embryo

Fully mature
 Two main sources of stem cells:

1. Embryonic Stem cells

 derived from a four- or five-day-old human embryo that is in the
blastocyst phase of development

2. Adult Stem Cells

 found in tissues such as the brain, bone marrow, blood, blood
vessels, skeletal muscles, skin, liver and umbilical cord.

https://www.medicalnewstoday.com/info/stem_cell
Blastocyst Diagram
Stem Cell Applications
• Tissue repair
- nerve, heart, muscle, organ, skin
• Cancers
• Autoimmune diseases
- diabetes, rheumatoid arthritis,
multiple sclerosis
 Technical Challenges

•Source - Cell lines may have mutations.

•Delivery to target areas
•Prevention of rejection
•Suppressing tumors
Why is Stem Cell Research So
Important to All of Us?

Stem cells can replace diseased or damaged cells

Stem cells allow us to study development and
genetics
Stem cells can be used to test different substances
(drugs and chemicals)
Why the Controversy Over Stem cells?
• Embryonic Stem cells are derived from extra
blastocysts that would otherwise be discarded
following IVF.
• Extracting stem cells destroys the developing
blastocyst (embryo).

-Questions for Consideration-

• Is an embryo a person?
• Is it morally acceptable to use embryos for
research?
• When do we become “human beings?”
Group Activity:

Creative Presentation on the topic chosen.

Maximum of 10 mins per group.

Topics:
1. What are the recent developments of gene therapy and
stem cell research?
2. What will be the future of gene therapy and stem cell
research?
3. What are the ethical and social issues concerning gene
therapy and stem cell research?
4. What are the risks and challenges of gene therapy and
stem cell research?