Gene

Therapy
BY: GROUP 2
 List of key concepts
- What is gene therapy

Our Agenda - Mechanism of gene therapy

- Types of gene therapy

for Today
- Identify and describes the delivery method.
- Advantages and disadvantages of gene therapy.
- Considering the ethical issue of Gene therapy
- Knowing the recent developments of Gene therapy
What is
Gene Therapy?
Gene therapy introduces genes into existing

cells to prevent or cure a wide range of

diseases.
It is a technique for correcting defective

genes responsible for disease development

The first approval of gene therapy was in

Europe in 2012, a rare enzyme deficiency

that causes pancreatitis but was

discontinued due to the high cost and low

demand.
Types of Gene Therapy

SOMATIC GENE THERAPY GERMLINE GENE THERAPY

Transfer of a section of DNA to any
Transfer of a section of DNA to cells that
cell of the body that doesn’t
produces eggs or sperm.[GERM CELLS]
produce sperm or egg.[SOMATIC
Genes introduced into eggs and
CELLS]
sperms.
Introduction of genes into bone
It is heritable and passed on for later
marrow cells, blood cells, skin cells
generations.
and etc.
For safety, ethical and technical
Will not be inherited by later
reasons, it is not being attempted at
generations.
present.
At present all researchers are
directed to correct genetic defects
in somatic cells.
 APPROACHES IN GENE THERAPY

IN VIVO GENE

EX VIVO GENE

THERAPY THERAPY
Direct delivery of genes into the

Transfer of genes to cultures

cells of a particular tissue in the

cells and reinsertion.

body.

EX VIVO THERAPY
“outside the body”
A cell that can be altered
outside the body and then
infused into the bloodstream
through a vein.
Process of Ex Vivo Gene Therapy
1. Isolate cells with genetic defects
2. Grow the cells in a culture
3. Introduce therapeutic genes
4. Select genetically corrected cells and grow
5. Transplant the modified cells to the patient

Process of Ex Vivo Gene Therapy in General

1. Remove cells from the patient
2. Add therapeutic genes to the cells
3. Return Cells back to the patient
An example of Ex Vivo Therapy is CAR T Cells
Therapeutics or also known as Chimeric Antigen
Receptor T Cells
CAR T Cells Therapeutics are T cells that have been
genetically engineered to produce an artificial T cell
receptor for use in immunotherapy.
Process of CAR T Cells Therapeutics
1. T cells are isolated from the patient's blood

2. A new gene encoding a chimeric antigen receptor is incorporated

into the T cells.

Process of CAR T Cells Therapeutics
3. Engineered T cells are now specific to the desired target

antigen

5. Engineered T cells are infused back into the patient

4. Engineered T cells are expanded in

tissue culture

IN VIVO THERAPY
In vivo gene therapy is a strategy in which
genetic material usually in the form of DNA, is
applied to modify the genetic repertoire of
target cells for therapeutic goals.
With in vivo gene therapy, the target cells or
tissues need to be easily accessible for
injection of the new genetic material.
·direct delivery of genetic material either
intravenously (through an IV) or locally to a
specific organ (ex: directly into the eye).
Process of In Vivo Gene Therapy

1. Copies of therapeutic genes are inserted into viral DNA, Liposomes, or in

form of Plasmid DNA.
2. Genetically altered DNA is inserted into the patient’s body by cell-specific
direct tissue injection.
3. Inside the body, the inserted DNA is incorporated into the cells of the
specific tissue it was injected into. These cells now encode and produce
the needed protein encoded by the inserted gene.
VECTORS OF GENE
THERAPY
To transfer the desired gene into a
target cell, a carrier is required. Such
vehicles of gene delivery are known
as Vectors.
Main classes
1. Viral vectors (adeno-associated
viral, adenoviral, lentiviral, retroviral)
2. Non- Viral vectos
 Why Viruses?

For gene therapy, there are four main types of

vectors, all derived from viruses because a
virus is good at getting into cells. But the viral
genes are removed, so only the therapeutic
genes are delivered. Researchers carefully
choose which viral vector to use to treat a
disease based on:
How well do researchers understand the
virus
How well the virus can target certain cells
How safe it is to use
VIRAL VECTORS
1. Adeno-Associated Viral (AAV) Vectors
Are able to deliver only small DNA packages or genes, to cells.
Are non-integrating, or don’t insert themselves into cells’ genome
Are typically used to target non-dividing cells, such as liver or nerve cells
Are most effective for in vivo treatments when vectors are injected directly into the
body Can be limited by innate immunity since many people are exposed to AAVs
through natural infections. The immune cells may destroy the vector before it can
deliver the therapy
Can be limited to a single dose since antibodies may develop
VIRAL VECTORS
2. Adenovirus Vector System
Can deliver packages up to 8x larger than AAVs
Are similar to AAVs in that they are non-integrating into the cells’ genome and target non-
dividing cells
Can cause strong immune responses resulting in potentially harmful inflammation throughout
the body, which decreases treatment effectiveness. Scientists are working to improve this
vector type to have a milder response

3. Lentiviral + Retroviral Vectors

Can deliver larger genetic packages of RNA, which is converted into DNA. During this process, the
vectors integrate into the genome of the target cell
Are typically used to target dividing cells, like T cells, which are immune cells, and stem cells. The new
genetic material is copied into all the new cells beyond the original cell
Are most effective for ex vivo treatments when the person's cells are removed from the body,
modified by vectors delivering working genes, and then returned to the body to improve function
Other Limitations
1. Off-target effects
when tissues other than the main target tissue may be affected after administration.
Vectors that insert their genetic package into the host genome can potentially integrate at an
incorrect location of the genome and cause unintended consequences.

2. Manufacturing
Creating a very large quantity of safe viral vectors requires time, effort, and resources.
The complexities of the process add to manufacturing costs and make it hard to effectively
streamline production.
Researchers will continue working towards more efficient production methods as more gene
therapies are researched and approved for use.
NON-VIRAL VECTORS

Researchers are also developing more non-viral vectors to

address (or eliminate) these limitations. Non-viral vectors
are cheaper to manufacture than their viral counterparts.
They can potentially deliver larger genetic packages, allow
for repeated dosing, and make quality control easier. Non-
viral vectors also have the benefit of a lowered chance of
triggering adverse immune responses.
METHODS OF GENE DELIVERY ( PHYSICAL )
1. Electroporation (EP)
It utilizes physical force to import therapeutic drugs and macromolecules, such as DNA
and proteins, from extracellular compartments into cells.
Electric pulses of different intensities and duration are applied according to the target
cell, tissue, or organ.

2. Gene Gun
Employs a high-pressure delivery system to shoot tissue with gold or tungsten
particles that are coated with DNA

3. Hydrodynamic Intravascular Injection

This approach combines naked DNA and hydrodynamic pressure generated by rapid
injection of a large volume of fluid into a blood vessel, to deliver genetic materials into
parenchyma cells.
METHODS OF GENE DELIVERY ( PHYSICAL )
4. Sonoporation
It is a non-viral gene delivery system for increasing plasmid DNA transfer across
biological cell membranes.

5. Microinjection
Process of using a glass micropipette to insert microscopic substances
into a single living cell.
Normally performed under a specialized optical microscope setup called
A micromanipulator.
METHODS OF GENE DELIVERY ( CHEMICAL )
1. USING DETERGENT MIXTURES
Certain charged chemical compounds like Calcium phosphates are mixed with
functional cDNA of the desired function.
The mixture is introduced near the vicinity of recipient cells.
The chemicals disturb the cell membrane, widen the pore size, and allow cDNA to
pass through the cell.

2. LIPOFECTION
It is a technique used to inject genetic materials into a cell by means of
liposomes.
Liposomes are artificial phospholipid vesicles used to deliver a variety of
molecules including DNA into the cells.
Advantages and

Disadvantages of Gene Therapy

ADVANTAGES
DISADVANTAGES
Long-lasting effects.
High cost.
Only needs to be given once.
Immune response to the transferred
Earlier undergo can be better.
gene stimulates a potential risk to
Potential to eliminate and prevent
heredity disease. gene therapy.
Positive effects of a gene therapy Potential for short-term efficacy.
are passed down through Experimental.
generations. Multigene disorders.
 Who will have access to such

therapy?

Is it against the beliefs of the

Christian Doctrine and God?

Ethical
Considerations Should people be allowed to use

gene therapy to modify basic

human traits such as intelligence,

height or athletic ability.?

Is it all right to use the therapy

in the prenatal stage in the

development of babies?
 How can the “good” and “bad”

uses of these technologies be

distinguished?

Who decides which traits are

normal and which constitute a

Ethical disability or disorder?

Considerations Will the high costs of gene

therapy make it available only
to the wealthy?

Could the widespread use of

gene therapy make society less
accepting of people who are
different?
RECENT DEVELOPMENTS IN GENE THERAPY
On January 28, 2021 the First hybrid gene therapy shows early promise in
treating long QT syndrome.
Oct. 10, 2022 - Researchers Improve Vehicle for Delivering Gene Therapies to
the Central Nervous System
October 19, 2022 - Researchers identify a gene as a potential target in
treatment-resistant brain cancer glioblastoma multiforme
November 3, 2022 - Gene therapy targeting overactive brain cells could
treat neurological disorders
November 9, 2022 - Silicon nanochip could treat traumatic muscle loss
Technology previously shown to change skin tissue into blood vessels and
nerve cells
CONCLUSION:

Theoretically, gene therapy is the permanent solution for genetic disease,

but it has several complexities. At its current stage, it is not accessible to
most people due to its huge cost.
A breakthrough may come anytime and a day may come when almost
every disease will have gene therapy. Because we believe that gene therapy
has the potential to revolutionize the practice of medicine.
THANK YOU FOR LISTENING!!