NATIONAL UNIVERSITY OF MEDICAL SCIENCES

Assignment No. 1
GENE THERAPY
Khadija Tariq

This document contains a detailed explanation, types, and application of gene therapy.
 Introduction
Gene therapy is the use of genetic material (nucleic acids) to cure or prevent diseases by
delivering genes to the patient’s targeted cells. This experimental technique will be used to cure
certain genetic disease, cancer and viral diseases in the near future. There are multiple
approaches to gene therapy including

 Replacing mutated gene

 Inactivating a mutated gene
 Introducing a new gene to fight the disease

A lot of gene therapy techniques are under clinical trials. These techniques are still very
dangerous and the ethical boundaries are still being discussed and decided around the world.

History of gene therapy

A brief history of gene therapy and its development is described below:

 In early 1960s experiments were conducted to figure if foreign DNA can be successfully
transferred into a mammalian cell and produce its products with it being inheritable. These
experiments were conducted on HPRT (hypoxanthine guanine phosphoribosyl transferase)
deficient and thymidine kinase deficient cells. But because of the absence of sufficient gene
transferring methods the experiments didn’t show consistent results
 In the mid-late 1960s it was confirmed that foreign genetic information can be successfully
transferred to the targeted cells. This was due to the use of papovaviruses SV40 and plyoma for
transforming cells. The integrated DNA was functional, expressive and inheritable
 Different approaches were suggested to transfer the therapeutic genes safely and effectively to
human cells some scientists suggested that artificially constructed pseudo viruses maybe used.
But no isolated genes were available at this time.
 In the 1970s the recombinant DNA technology finally provide both the vector to transfer the
required gene and specific genes in cloned forms experimental support was given to this notion
by the preparation of a recombinant SV40 vector which successfully transferred foreign
sequences to mammalian cells.
 At this time the techniques were still not efficient enough to be used to carry out human
applications but these techniques still lay the basis of gene therapy
 In 1979 Martin Cline performed clinical trials on thalassemia patients which were heavily
criticized on ethical grounds
 In 1981-2 the use of retroviral vectors was proved with close to 100% efficiency.
 In 1983 Banbury conference was held in new york discussing the techniques on gene therapy
 In 1989-90 the first human clinical trial was approved for therapeutic studies.
 Types of gene therapy
There are two types of gene therapies depending on which type of cell is being treated, somatic or germ
line cells.

Somatic gene therapy

It is the transfer of foreign DNA to any type of cell that doesn’t produce sperm or eggs. The results of the
treatment are not transferred to the next generation. This type of gene therapy is accepted worldwide and
has clinical trials in progress. The draw backs to this type of therapy include the short lived effects of the
treatment and an overall requirement of multiple treatments. The second problem that arises is the
targeted supply of the gene to specific tissues. Now, there are two main methods for somatic gene therapy

I. In vivo gene transfer ( the genes are incorporated in the cells inside the body )
II. Ex vivo gene transfer (the specific cells are transformed outside the body and then targeted to
specific tissues )

Somatic gene therapy is being used to help patients with diseases such as muscle dystrophy, cystic
fibrosis, cancer and certain infectious diseases.

Germ line gene therapy

It is carried out when the foreign gene is transferred into the cells that produce eggs or sperms the results
of the treatment are transferred onto the next generations. This type of gene therapy produces permanent
effects. This type of treatment is usually done during in-vitro fertilization or during earlier embryonic
stages. This type of gene therapy is banned in a lot of countries around the world because the long term
effects of such procedures are unknown and can be harmful for future generations. However, this type of
therapy can be used to delete certain disease causing genes from families and then eventually from the
population. This type of gene therapy can be used to cure Alzheimer and other neurological diseases.

Vectors
Gene transfer into cells can be done by two mainly different types of methods which are viral and non-
viral methods.

Viral vectors

Viruses infect their host cells and incorporate their genetic material into that of the host to use its
metabolic machinery as a part of their life cycle. Now scientists use these viruses to transfer their desired
genes into the human cells. This method requires cutting out the disease causing genes in the virus and
adding the desired gene. This whole process is done through genetic engineering. In the beginning
retroviruses were used to perform this function but later on adenoviruses, herpes virus and some other
viruses were also used in clinical trials. Different viruses are classified on the basis of

 Infection
 Host genome interaction
  Transgene expression
 Packaging capacity

There are also drawbacks to using viruses as vectors such as some viruses target a spectrum of hosts
instead of just one type of cells so they can transfer the therapeutic gene to normal body cells as well
which can cause multiple problems.

Another major risk factor is the insertion of new gene at a wrong location in the DNA which can lead to
serious issues including cancer. This side effect has been seen in clinical trials of X-SCID patients
causing them leukemia

Other concerns include the overexpression of the gene to a level that can be very harmful.

Non-viral vectors

There are certain methods in which no viruses are used to transmit genetic material into host cells. These
methods use either direct DNA or other complex molecules to transform host cells some of the methods
are discussed below:

Naked DNA is the simplest method of transfection. There are different ways by which naked DNA is
transferred to the host cells one of the simplest being transmission by plasmid injections. Other
techniques include ‘gene guns’ which shoot gold coated DNA at the target cells at high pressures

Oligonucleotides are molecular complexes that are used to stop the expression of a faulty gene. These
complexes bind to the transcription factor and acts as a decoy to stop the transcription of the faulty gene
lowering the expression and essentially preventing the disease.

Lipoplexes are lipid DNA complexes in which the anionic DNA is packed inside a cationic lipid making
a structure like micelle or liposome. These complexes prevent the degradation of DNA inside a cell
before it’s incorporated into the host genome. In recent studies these Lipoplexes are shown to transfect
respiratory epithelial cells so they may be used as a cure for cystic fibrosis in the future.

Applications

 Other than disease therapy one of the applications of gene therapy is gene doping. The use of
gene therapy to restore lost muscle due to age or disease is gene doping. Specialists are concerned
that athletes might misuse this technology to enhance performance. Mainly because this type of
gene therapy can be caught through testing, there are organizations working against gene doing in
athletes the most important one being WADA (world anti-doping agency).
 Genetic engineering to enhance human features is a long debated ethical issue exactly due to
which germ line gene therapy is banned in a lot countries around the world. Gene therapy can be
used to enhance intelligence, memory and even physical features etc.
References
http://www.genetherapynet.com/

Wirth, Thomas, Nigel Parker, and Seppo Ylä-Herttuala. "History of gene therapy." Gene 525.2 (2013):
162-169.

Friedmann, Theodore. "A brief history of gene therapy." Nature genetics 2.2 (1992): 93-98.