GENE

THERAPY

Gene Therapy
It is a technique for correcting defective

genes that are responsible for disease

development
There are four approaches:
1.
2.
3.
4.

A normal gene inserted to compensate

for a nonfunctional gene.
An abnormal gene traded for a normal
gene
An abnormal gene repaired through
selective reverse mutation
Change the regulation of gene pairs

The Beginning
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

Scientist took the logical step of trying to introduce genes straight

into human
cells, focusing diseases caused by single-gene defects, such as cystic
fibrosis,
hemophilia, muscular dystrophy and sickle cell anemia. However this
has been
much harder than modifying simple bacteria, primarily because of the
problems
involved in carrying large section of DNA and delivery it to the right
site on the
genome

The First Case

The first gene therapy was performed on

September 14th, 1990 at US national

institute of health.

Ashanti DeSilva was treated for SCID

Severe 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

1.

Gene therapy utilizes two

theoriticaly
: line
1..Germ
line genepossible
therapy :In directions
the case of germ

gene therapy, germ cells, i.e., sperm or eggs, are

modified by the introduction of functional genes,
which are ordinarily integrated into their genomes.
Therefore, the change due to therapy would be
heritable and would be passed on to later
generations. This new approach, theoretically, should
be highly effective in counteracting genetic disorders
and hereditary diseases.
2. Somatic Gene Therapy :In the case of somatic gene
therapy, the therapeutic genes are transferred into
the somatic cells of a patient. Any modifications and
effects will be restricted to the individual patient
only, and will not be inherited by the patient's
offspring or later generations

 SOMATIC GENE THERAPY IS SUBDIVIDED INTO

AUGMENTATION GENE THERAPY

TARGETED GENE THERAPY

In augmentation gene therapy the

functional gene is introduced in to the
somatic cell containing defective gene. Hence
the modified cell contain both defective gene
and newly introduced functional gene.
The targeted gene therapy abnormal gene
is replaced by a functional gene using
recombinant DNA technology.

Vectors used in
gene therapy :
mainly viruses

Why Viruses ?
A virus is the simplest organism there isit is pretty much just
genetic material wrapped up in a protein coat.
a virus cant live on its own
it survives and multiplies by parasitically attacking living cells
and injecting its genetic material into cells.
The fact that many viruses are cell specific (e.g., a certain
virus may only infect heart cells or lung cells) helps scientists
target just the desired cells.

That makes it an ideal mechanism for getting genes into a cell.

a scientist might take the damaging, infecting portion of the

DNA out of the virus and add to it the desired gene segment.

Some of the different types of viruses as

gene therapy vectors :
1. Retroviruses
2. Adenoviruses
3. Adeno-associated viruses
4. Herpes simplex viruses

Retroviruses
Created double stranded DNA copies from RNA genome
The retrovirus goes through reverse transcription
using reverse transcriptase and RNA
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 genes code (disrupted cell
division causes cancer from uncontrolled cell
division)

vectors used are derived from the human

immunodeficiency virus (HIV) and are being
evaluated for safety.
May trigger immune response .

Retroviruses carrying healthy gene are mixed with

unhealthy cells taken from a patient.
Retroviruses infect unhealthy cells with healthy gene,
adding the gene to patients DNA.
Healthy cells then injected back to the patient.

Adenoviruses
Adenoviruses carry their genetic material in the form of DNA.
When these viruses infect a host, they introduce their DNA

molecule into the host but not incorporated into the host
genetic material.
These extra genes are not replicated, so when the host undergo

cell division, the descendants of the cell will not have the extra
gene.
This means that treatment with adenovirus will require regular
doses to add the missing gene every time.
Adenoviruses can invade slower dividing cells, such as lung

cells
Like retroviruses, immune response may reduce its
effectiveness

Adenovirus cont

http://en.wikipedia.org/wiki/Gene_therapy

Adeno-associated Viruses
Adeno-associated viruses are believed to occur naturally in

humans, existing without causing disease or instigating in

immune response from the body. This type of virus is being
used, because it is non-pathogenic (most people carry this
harmless virus).
AAV are small viruses with a genome of single stranded

DNA. They insert their material specifically into chromosome

19, at a specific site.
Disadvantages in using AAV are the small amount of DNA
it can carry (only 2 genes) and the difficulty in producing it.

Continued

Several trials with AAV are ongoing or in

preparation, mainly trying to treat muscle and
eye disease, the two tissues where the virus
seems particularly useful.

hemophilia treatments, for example, a genecarrying 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

Herpes Simplex Viruses

Herpes simplex viruses : A class of

double stranded DNA virus that usually

used to target the nervous system
because herpes simplex virus usually
infects neurons.
Ex. Herpes simplex virus type 1

NON-VIRAL OPTIONS
Direct introduction of therapeutic

DNA

Intramuscular injection of a naked

DNA plasmids has been used to
transport normal functional
gene in to a cell.
Similarly
oligonucleotides, lipoplexes,
polyplexes are also used as a
vector in gene therapy.

What diseases could be treated

with
About 4000 diseases have been traced
gene
therapy
to gene disorders.
Current and possible candidates for gene

therapy include cancer, AIDS, cystic

fibrosis, Parkinsons, Alzheimers disease,
Retinoblastoma, cardiovascular disease
and arthritis.
more than 600 gene therapy clinical trials
were under way in the US but only a
handful of these are in advanced stages.

What factors have kept gene

therapy from becoming an
Short-lived nature of gene therapy.
effective
treatment
for
genetic
Immune response .
disease
? viral vector : viruses may present
Problem with
a variety of potential problems to the patient
toxicity, immune and inflammatory response
and also cause disease.
Multigene disorder : multigene or multifactorial

disorders would be difficult to treat effectively

using gene therapy.
High risk factor .

What risk are associated with

current
gene
Viruses can
usually infect
more than
one type of cell,
therapy
trials
?
thus they might infect healthy cells.

Another danger is that the new gene might be inserted in

the wrong location in the DNA, possibly causing cancer

or harmful mutations to the DNA.
There is a slight chance that the DNA could unintentionally
be introduced into the patients reproductive cells, so it
may be passed on if the patient has children after
treatment.
The possibility that transferred genes could be
overexpressed , producing so much of the missing
protein as to be harmful .
the viral vector could cause inflammation or an immune
reaction ; and that the virus could be transmitted from
the patient to other individuals or into the environment.

 RELIGIOUS CONCERN: Most of the

people believe that human are created

by god and alteration of human gene
is equal to interfering or corrupting
gods work..

The Future of Gene Therapy

Current uses of gene therapy focus on

treating or curing existing conditions. In
the future, the focus could shift to
prevention. As more of the human
genome is understood, medicine will know
more about which genes contribute to or
cause disease. With that knowledge in
hand, gene therapy could be used to head
off problems before they occur.

Gene therapy and gene-based

medicine will revolutionize
medicine over the next ten to
twenty years. The big question is
when.

Gene therapy pioneer French Anderson, 2001

http://www.wellesley.edu/Biology/Courses/219/Gen_news/i3_Gene_Therapy.jpg

Thank You