Introduction

Gene therapy as it is used to treat medical conditions and the benefits and challenges it raises.
In summary Gene therapy is the transfer of a therapeutic or working copy of a gene into specific cells of
an individual The technique may be used to replace a faulty gene, or to introduce a new gene whose
function is to cure or to favourably modify the symptoms of a condition. Gene therapy is still an
experimental technique and much research remains to be done before this approach to the treatment
of conditions will realise its full potential. Gene therapy involves the transfer of a therapeutic or working
gene copy into specific cells of an individual. It may be used to:  Replace a faulty gene  Introduce a
new gene whose function is to cure or to favourably modify the clinical course of a condition  Inactivate
or “knock out” a faulty gene that is not functioning properly. The potential of gene therapy is very
broad, with research involving a number of diseases such as severe combined immune-deficiencies
(SCID), haemophilia, Parkinson’s disease, many forms of cancer and HIV.

This is probably an important advancement in stem cell research, since it allows researchers to
obtain pluripotent stem cells, which are important in research, without the controversial use of
embryos. There were two main issues concerning stem cell research with both pros and cons: How the
knowledge will be used Concerns about the methods.The first issue is really not just about stem cell
research, as it may be applied to most research about human health. Since 2007, the second point,
concerns about the methods involved, has been less debated, because of scientific developments such
as iPS

There are many challenges to successful gene therapy. Firstly, the condition in question must be
well understood and the underlying causative gene identified. A working copy of the gene involved must
be available and the specific cells in the body requiring treatment must be identified, accessible and a
means of efficiently delivering working copies of the gene to these cells must be available. Of all these
challenges, the one that is most difficult is the problem of gene delivery i.e. how to get the new or
replacement genes into the desired tissues. Some of the vectors for delivering the working copy of the
gene to the target cells include using: a) Harmless viruses One of the most promising methods currently
being developed is the use of harmless viruses that can be used to carry genes into cells. Scientists now
have the knowledge and skills to remove the virus’ own genes and to replace them with working human
genes. These altered viruses can then be used to deliver genes into cells with great efficiency. When
viruses are used in this way they are known as vectors. Some of these vectors are capable of not only
carrying the gene into the cell but also of inserting the gene into the genetic material of the cell. Once in
the right location within the cell of an affected person, the transplanted gene is switched on. The
transplanted gene can then issue the instructions necessary for the cell to make the protein that was
previously missing or altered. b) Stem cells Another technique with potential is the use of stem cells in
delivering gene therapy. Stem cells are cells that have not yet differentiated into a specific tissue or
organ cell. In this technique, stem cells are manipulated in the laboratory in order to make them accept
new genes that can then change their behaviour. For example, a gene might be inserted into a stem cell
that could make it better able to survive chemotherapy. This would be of assistance to those patients
who could benefit from further chemotherapy following stem cell transplantation.
Problem

Gene therapy is used to correct defective genes in order to cure a disease or help your body better
fight disease.

Researchers are investigating several ways to do this, including:

 Replacing mutated genes. Some cells become diseased because certain genes
work incorrectly or no longer work at all. Replacing the defective genes may help
treat certain diseases. For instance, a gene called p53 normally prevents tumor
growth. Several types of cancer have been linked to problems with the p53 gene. If
doctors could replace the defective p53 gene, that might trigger the cancer cells to
die.

 Fixing mutated genes. Mutated genes that cause disease could be turned off so
that they no longer promote disease, or healthy genes that help prevent disease
could be turned on so that they could inhibit the disease.

 Making diseased cells more evident to the immune system. In some cases,
your immune system doesn't attack diseased cells because it doesn't recognize
them as intruders. Doctors could use gene therapy to train your immune system to
recognize the cells that are a threat.

 Introducing a new gene into the body to help fight a disease.

Solution:

A healthy gene is inserted into a carrier, called a vector, and transferred to the affected cells,
either inside or outside the body.
There are mainly two approaches for the transfer of genes in gene therapy: 1. Transfer of genes into
patient cells outside the body (ex vivo gene therapy) 2. Transfer of genes directly to cells inside the body
(in vivo).

Ex vivo gene therapy  In this mode of gene therapy genes are transferred to the cells grown in culture,
transformed cells are selected, multiplied and then introduced into the patient.  The use of autologous
cells avoids immune system rejection of the introduced cells.  The cells are sourced initially from the
patient to be treated and grown in culture before being reintroduced into the same individual.  This
approach can be applied to the tissues like hematopoietic cells and skin cells which can be removed
from the body, genetically corrected outside the body and reintroduced into the patient body where
they become engrafted and survive for a long period of time.  Figure 8-1.5.1 shows a self-explanatory
schematic diagram for ex vivo gene transfer.

In Vivo Gene Therapy  In vivo method of gene transfer involves the transfer of cloned
genes directly into the tissues of the patient.  This is done in case of tissues whose individual
cells cannot be cultured in vitro in sufficient numbers (like brain cells) and/or where re-
implantation of the cultured cells in the patient is not efficient.  Liposomes and certain viral
vectors are employed for this purpose because of lack of any other mode of selection.  In case
of viral vectors such type of cultured cells were often used which have been infected with the
recombinant retrovirus in vitro to produce modified viral vectors regularly. These cultured cells
will be called as vector-producing cells (VPCs)). The VPCs transfer the gene to surrounding
disease cells.  The efficiency of gene transfer and expression determines the success of this
approach, because of the lack of any way for selection and amplification of cells which take up
and express the foreign gene.
Results

The possibilities of gene therapy hold much promise. Clinical trials of gene
therapy in people have shown some success in treating certain diseases, such as:

 Severe combined immune deficiency

 Hemophilia

 Blindness caused by retinitis pigmentosa

 Leukemia
Your specific procedure will depend on the disease you have and the type of gene
therapy being used.

For example, in one type of gene therapy:

 You may have blood drawn or you may need bone marrow removed from your
hipbone with a large needle.

 Then, in a lab, cells from the blood or bone marrow are exposed to a virus or
another type of vector that contains the desired genetic material.

 Once the vector has entered the cells in the lab, those cells are injected back into
your body into a vein or into tissue, where your cells take up the vector along with
the altered genes.

Gene therapy has some potential risks. A gene can't easily be inserted directly into your
cells. Rather, it usually has to be delivered using a carrier, called a vector.

The most common gene therapy vectors are viruses because they can recognize
certain cells and carry genetic material into the cells' genes. Researchers remove the
original disease-causing genes from the viruses, replacing them with the genes needed
to stop disease.

This technique presents the following risks:

 Unwanted immune system reaction. Your body's immune system may see the
newly introduced viruses as intruders and attack them. This may cause
inflammation and, in severe cases, organ failure.

 Targeting the wrong cells. Because viruses can affect more than one type of
cells, it's possible that the altered viruses may infect additional cells — not just the
targeted cells containing mutated genes. If this happens, healthy cells may be
damaged, causing other illness or diseases, such as cancer.

 Infection caused by the virus. It's possible that once introduced into the body,
the viruses may recover their original ability to cause disease.

 Possibility of causing a tumor. If the new genes get inserted in the wrong spot in
your DNA, there is a chance that the insertion might lead to tumor formation.
Recommendations

 Targeting the correct cells

 Reducing the risk of side effects

References:

 uniQure. http://www.uniqure.com/pipeline/clinical-programs/. Accessed July 2016.

 What is gene therapy? http://www.genetherapynet.com/types-of-gene-therapy.html.
Accessed July 2016.
 Gene Therapy and Emerging Molecular Therapies (Elsevier 2005); Chpt 5:pp 50.
 Salmon F, Grosios K, Petry H. Expert Rev Clin Pharmacol. 2014;7(1):53–65.
 Scott LJ. Drugs. 2015;75(2):175–82.
 D’Avola D, et al. J Hepatol. doi: http://dx.doi.org/10.1016/j.jhep.2016.05.012
 https://l.facebook.com/l.php?u=https%3A%2F%2Fghr.nlm.nih.gov%2Fprimer%2Ftherap
y%2Fgenetherapy%3Ffbclid%3DIwAR0S2WwSCA4eGSzFPHXU32x6UsOvwpxv0vBtj
CVGgXrnoFrckQVYG3GyBOA&h=AT3ynt_88IAquf387qX58oAsCVHdM7QfMZEw5
NzQT1OtXulJ4mYfzpV-xGpK0FUH5JLIL5-P2ASLrtoNoEKw1dH0aukaIYkIb75-
hU9ELzgZ9hyzYNNrVB02voZPhuyLBx_Y
 https://l.facebook.com/l.php?u=https%3A%2F%2Fwww.thehastingscenter.org%2Fbriefi
ngbook%2Fstem-
cells%2F%3Ffbclid%3DIwAR2f_pc6XYsnZEusloGep1Aeq23EPl1MBJjQc4633jCtsCdL
m_A_jsBtD3Q&h=AT3ynt_88IAquf387qX58oAsCVHdM7QfMZEw5NzQT1OtXulJ4m
YfzpV-xGpK0FUH5JLIL5-P2ASLrtoNoEKw1dH0aukaIYkIb75-
hU9ELzgZ9hyzYNNrVB02voZPhuyLBx_Y
 https://learn.genetics.utah.edu/content/stemcells/scissues/
 AR Prabhakar (2011). Gene Therapy and its Implications in Dentistry. International Journal of
Clinical Pediatric Dentistry 4(2):85-92 Cavazzana-Calvo M., Hacein-Bey S., de Saint Basile G.,
Gross F., Yvon E., Nusbaum P., Selz F., Hue C., Certain S., Casanova J.L., Bousso P., Deist F.L.,
Fischer A.(2000). ‘Gene therapy of human severe combined immunodeficiency (SCID)-X1
disease’. Science 288:669. Chris Mason, Peter Dunnill (2008). A brief definition of regenerative
medicine: Editorial. Regenerative Medicine, 2008, 3(1), 1–5 [47] Gottesman M. (2003). “Cancer
gene therapy: An awkward adolescence”. Cancer Gene Therapy 10: 501.