Gene Therapy

What Are Genes?

• Genes are carried on chromosomes

and are the basic physical and
functional units of heredity.
• Genes are specific sequences of
nucleotides that encode instructions
on how to make proteins.
• When genes are altered so that the
encoded proteins are unable to carry
out their normal functions, genetic
disorders result.
 What Are Genetic Disorders?

• Genetic disorder is a disease caused

by a "variation" or "mutation“ of a
gene.
• Genetic disorders can be passed on
to family members who inherit the
genetic abnormality.
• A small number of rare disorders
are caused by a mistake in a single
gene.
• Most disorders involving genetic
factors, such as heart disease and
most cancers, arise from a interplay
of multiple genetic changes and
environmental factors.
 Gene Therapy

 Gene therapy is a technique for correcting

defective genes responsible for disease
development.

 Gene therapy is the use of genes as medicine‟. It

involves the transfer of a therapeutic or working
gene copy into specific cells of an individual in
order to repair a faulty gene copy.
 Gene Therapy

Gene Therapy is the insertion,

alteration or removal of genes
within an individual cell’s and
biological tissues to treat disease.
It involves adding a normally
functioning copy of the gene(s) to
enough affected cells to restore
normal function.
 Why Gene Therapy?

• Gene therapy can be used for a

number of diseases, such as severe
combined immune-deficiencies,
hemophilia, Parkinson's disease,
cancer and even HIV through a
number of different approaches.
• This technique may allow doctors
to treat a disorder by inserting a
gene into a patient’s cells instead of
using drugs or surgery.
 Gene Therapy - Background
• 1990 - The first gene therapy journal
published, Human Gene Therapy.
• 1990 - The first approved gene therapy
clinical trial took place when Ashanthi
De Silva, a 4 year old girl with ADA-
deficient Severe Combined
Immunodeficiency, was given her own
T cells engineered with a retroviral
vector carrying a normal ADA gene.
• 2000 - The first gene therapy cure was
reported when Alain Fischer (Paris)
“Bubble Boy”
succeeded in totally correcting children
with SCID-X1, or “bubble boy”
syndrome.
 Basic Processes of Gene Therapy
To design and carry out a gene therapy
treatment, a researcher must:
1. Identify the gene(s) responsible for the
disorder.
2. Make copies of the normal gene.
3. Insert the copies into vectors.
4. “Infect” the affected cells with the vectors.
5. Activate the gene so that transcription and
translation take place.
• In general, a gene cannot be directly inserted
into a person’s cell. It must be delivered to the
cell using a carrier, or vector. Vectors are
carrier molecules which are employed to
enhance gene transfer efficiency in gene
therapy. In optimizing a particular vector, one
must consider:
– Host immune response.
– Must target specific tissues for long term gene
expression.
– Regulation of the gene after insertion.
 Viral Vector Carrying Healthy Gene

Cell with mutated Vector inserts New gene replaces

gene(s) healthy gene into the defective genes
cell
 Types of Gene Therapy
• Germ line Gene Therapy: It involves permanent
transfer of the gene to the sperm or egg cells. Gene
therapy using germ line cells results in permanent changes
that are passed down to subsequent generations. germ line
therapy is not permitted in any county, on the basis that it
is unethical.

• Somatic cells Gene Therapy: It ideally involves the

transfer of genes to the affected cells. Somatic cell therapy
is viewed as a more conservative, safer approach because
it affects only the targeted cells in the patient, and is not
passed on to future generations.
 Somatic Cell Gene Therapy
Two Types:
• In vivo gene therapy: delivery of new genetic
material directly to target cells within the body
– The challenge lies in ensuring the specificity and in
reaching the correct target cells within the body
• Ex vivo therapy: target cells are removed from the
body and then genetically modified
– The cells are then returned to the body after selection
and amplification
– This is a safe method but dependent on the type of cells
being targeted
 Approaches to Gene Therapy
Several approaches to gene therapy are being
tested, including:

 Replacing a mutated gene that causes disease

with a healthy copy of the gene.
 Inactivating, or “knocking out,” a mutated
gene that is functioning improperly.
 Introducing a new gene into the body to help
fight a disease
 Viruses as Vectors

• Currently, the most common type of vectors

are viruses that have been genetically altered
to carry normal human DNA.
• 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.
 Viral Vectors
• The viral vectors increases the capability of
viruses to transfer genetic material into infected
cell.
• Viruses insert their DNA into cells with high
efficiency.
• Viruses are evolved by genetic modification of
 Adenovirus
 Adeno-associated virus(AAV)
 Retroviruses
 Lentivirus
 ADENOVIRUS AS A VECTOR IN GENE
THERAPY
• The Adenovirus is ubiquitous- it has
been isolated from a large number of
different species with over 100 known
serotypes.
• Can rapidly infect a large range of
human cells.
• Low pathogen city in humans.
• Can hold large segments of DNA.
• Genome does not undergo
rearrangement at high rates.
• DNA is easy to manipulate with current
recombinant DNA techniques.
 Cancer
 Multiple gene therapy strategies have been
developed to treat a wide variety of cancers,
including suicide gene therapy, anti-angiogenesis
and therapeutic gene vaccines.

Two-thirds of all gene therapy trials are for

cancer and many of these are entering the
advanced stage e.g. gene vaccine trials for
prostate cancer and pancreas cancer.