Gene Therapy

Case studies

SUKESH POTLA 27 March 2012

Agenda or Summary Layout

Topics to be discussed
Genes and Genetic disorders Gene therapy Limitations and Problems of Gene therapy Regulations Discussion

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Genes

What can they do ?

Genes are

carried on chromosomes

basic physical and functional units of heredity.

specific sequences of bases that encode instructions on how to make proteins.

Its the proteins that perform most life functions and even make up the majority of cellular structures.

WhyGenetic disorders occur ?

When genes are altered so that the encoded proteins are unable to carry out their normal functions, genetic disorders can result.

What is Gene Therapy ?

Why is it used ?

It is a technique for correcting defective genes that are responsible for disease development

Genes are Medicine?

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. GT is used to correct a deficient phenotype so that sufficient amounts of a normal gene product are synthesized to improve a genetic disorder

How is Gene Therapy carried out ?

Modification of somatic cells by transferring desired gene sequences into the genome. There are Four approaches

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

Delivery of Gene sequences

Two types of delivery systems
In vivo

Ex vivo

delivery of genes takes place in the body In vivo techniques usually utilize viral vectors Virus = carrier of desired gene Virus is usually crippled to disable its ability to cause disease Many viral vectors can stable integrate the desired gene into the target cells genome

delivery takes place out of the body, and then cells are placed back into the body Ex vivo manipulation techniques Electroporation Liposomes

www.wix.com/greenapple99/gene-therapy.jpg

Viruses used as delivery tools

These viruses are inactivated already

So, they are not able to reproduce

RNA viruses

Adenoviruses

Herpes viruses

DNA viruses

Retro viruses

www.biochem.arizona.edu/classes/bioc471/pages/.../Lecture25.1jpg

Direct delivery

Therapeutic gene is packed into retrovirus and injected into patient

Cell based delivery

Therapeutic gene is packed into retrovirus and injected into adult stem cells and injected into patient

Ex vivo delivery
types

Electroporation

Liposomes

www.biochem.arizona.edu/classes/bioc471/pages/.../Lecture25.2jpg

Electroporation

Liposomes

http://mol-biol4masters.masters.grkraj.org/html/Genetic_Engineering4C-TransformationAnimal_Cells_files/image002.gif

http://upload.wikimedia.org/wikipedia/en/thumb/2/28/Liposome.jpg/2 50px-Liposome.jpg

Limitations of Gene therapy

Gene

delivery

Limited tropism of viral vectors Dependence on cell cycle by some viral vectors (i.e. mitosis required)

Duration

of gene activity

Non-integrating delivery will be transient (transient expression)

Integrated delivery will be stable

Patient

safety

Immune hyper-responsiveness (hypersensitivity reactions directed against viral vector components or against transgenes expressed in treated cells)

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control/regulation

Gene

Most viral vectors are unable to accommodate full length human genes containing all of their original regulatory sequences Human cDNA often used much regulatory information is lost (e.g. enhancers inside introns)

Expense

Costly because of cell culturing needs involved in ex vivo techniques

Problems with Gene therapy

Short Lived

Hard to rapidly integrate therapeutic DNA into genome and rapidly dividing nature of cells prevent gene therapy from long time Would have to have multiple rounds of therapy

Immune Response

new things introduced leads to immune response increased response when a repeat offender enters

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Viral Vectors

patient could have toxic, immune, inflammatory response

also may cause disease once inside

Multigene Disorders

Heart disease, high blood pressure, Alzheimers, arthritis and diabetes are hard to treat because you need to introduce more than one gene May induce a tumor if integrated in a tumor suppressor gene because insertional mutagenesis

Gene therapy regulations

Who controls the use of Gene therapy ? Currently gene therapy is regulated by the US Food and Drug Administration (FDA) and the National Institute of Health (NIH). The FDA looks at safety and control measures associated with gene therapy. The NIH has a subcommittee, the Recombinant DNA Advisory Committee (RAC) that reviews gene therapy protocols.

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Recombinant DNA-Advisory Committee

Provides advice concerning advances in:

Recombinant technology New organisms under investigation Public attitudes associated with research in molecular biology

Examines clinical trials that involve the transfer of recombinant DNA to humans

All trials funded by NIH are registered with the RAC

Advisory to the Director NIH

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Before any human gene transfer trials can take place, the RAC reviews proposals and comments on the feasibility, risks, etc.

Safety concerns

Toxicity Contaminants Safe vectors? , Side effects

First Approved Gene Therapy

On September 14, 1990 at the U.S. National Institutes of Health, W. French Anderson M.D. and his colleagues R. Michael Blaese, M.D., C. Bouzaid, M.D., and Kenneth Culver, M.D., performed the first approved gene therapy procedure on four-year old Ashanthi DeSilva. Born with a rare genetic disease called severe combined immunodeficiency (SCID)

What about in Canada ?

Which government involves in Gene regulations ?

Health Canada's Biologics and Genetic Therapies Directorate (BGTD) is the Canadian federal authority that regulates biological drugs (products derived from living sources) and radiopharmaceuticals for human use in Canada, whether manufactured in Canada or elsewhere.

Before gene therapy can be used to treat a certain genetic condition or disorder, certain requirements need to be met:
The

faulty gene must be identified and some information about how it results in the condition or disorder must be known so that the vector can be genetically altered for use and the appropriate cell or tissue can be targeted. gene must also be cloned so that it can be inserted into the vector. the gene is transferred into the new cell, its expression (whether it is turned on or off) needs to be controlled

The

Once

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The balance of the risks and benefits of gene therapy for the condition or disorder must compare favourable to other available therapies. Sufficient data from cell and animal experiments are needed to show that the procedure itself works and is safe.

Discussion

How can good and bad uses of gene therapy be distinguished?

Although gene therapy could have potentially huge benefits to truly cure diseases forever, there are several factors that need to be considered. One factor that must be addressed is that future generations will forever be affected by our decisions.

Who decides which traits are normal and which constitute a disability or disorder?

that it is the individual's decision to make such decisions concerning their lives. What must be kept in mind is that the purpose of regulation is for the safety of individuals.

Will the high costs of gene therapy make it available only to the wealthy?

Problems like only the wealthy having access to expensive gene therapy treatments have already presented problems while problems such as genetic alterations becoming a luxury instead of a lifesaving treatment could become important.

Could the widespread use of gene therapy make society less accepting of people who are different?

Surely, when gene therapy is widespread, society gets adapted to people who are good in all qualities and this can lead to less acceptance of people who are different.

Should people be allowed to use gene therapy to enhance basic human traits such as height, intelligence, or athletic ability?

The problems seem but limitless when one starts thinking about certain people having access to becoming more intelligent or stronger through gene therapy.

References

http://www.ndsu.edu/pubweb/~mcclean/plsc431/st udents/ross. http://www.nserc-crsng.gc.ca/studentsetudiants/pd-np/laboratories-laboratoires/hcsc_eng.asp http://www.hc-sc.gc.ca/sr-sr/biotech/aboutapropos/gen_therap-eng.php