1. HISTORY AND DEVELOPMENT OF GENE THERAPY 1960: The concepts of Gene Therapy was introduced 1970: Friedmann and Roblin author of a paper in Science titled "Gene therapy for human genetic disease?” cite the first attempt to perform gene therapy 1990:  The first approved gene therapy case at the National Institute of Health, U.K. It was performed on a four year old girl named Ashanti DaSilva. It was a treatment for a genetic defect that left her with an immune system deficiency  New gene therapy approach repairs errors in messenger RNA derived from defective genes. This technique has the potential to treat the blood disorder Thalassaemia, Cystic fibrosis, and some cancers  Sickle cell disease is successfully treated in mice

2. 1992: Doctor Claudio Bordignon working at the Vita-Salute San Raffaele University, Milan, Italy performed the first procedure of gene therapy using hematopoietic stem cells as vectors to deliver genes intended to correct hereditary diseases 1999: Death of Jesse Gelsinger in a gene-therapy experiment resulted in a significant setback to gene therapy research in the United States 2006: Scientists at the National Institutes of Health (Bethesda, Maryland) have successfully treated metastatic melanoma in two patients. This study constitutes one of the first demonstrations that gene therapy can be effective in treating cancer. 2007- 2011: Research is still ongoing and the number of diseases that has been treated successfully by gene therapy increases. Retinal disease Colour blindness Adrenoleukodystrophy 2011: Medical community accepted that it can cure HIV as in 2008, Gero Hutter has cured a man from HIV using gene therapy

3. WHAT IS GENE THERAPY ? Definiton: an experimental technique for correcting defective genes that are responsible for disease development The most common form of gene therapy involves inserting a normal gene to replace an abnormal gene Other approaches used: RReplacing a mutated gene that causes disease with a healthy copy of the gene. IInactivating, or “knocking out,” a mutated gene that is functioning improperly. IIntroducing a new gene into the body to help fight a disease.

4. Researchers are studying gene therapy for a number of diseases, such as  Severe combined immuno-deficiencies (SCID) HHemophilia PParkinson's disease CCancer HHIV

6.  Result in permanent changes.  Potential for offering a permanent therapeutic effect for all who inherit the target gene.  Possibility of eliminating some diseases from a particular family.  Also raises controversy:  Some people view this type of therapy as unnatural, and liken it to "playing God”.  Others have concerns about the technical aspects.

8.  Affects only the targeted cells in the patient, and is not passed to future generations.  Short-lived because the cells of most tissues ultimately die and are replaced by new cells.  Transporting the gene to the target cells or tissue is also problematic.  Appropriate and acceptable for many disorders, including cystic fibrosis, muscular dystrophy, cancer, and certain infectious diseases.

12. BASIC PROCESS OF GENE THERAPY 1. VIRAL VECTOR 2. NON VIRAL VECTOR

13. GT utilizes the delivery of DNA into cells, which can be accomplished by a number of methods. The two major classes of methods :  recombinant viruses – VIRAL VECTOR  naked DNA or DNA complexes – NONVIRAL VECTOR

14. VIRAL VECTOR Viruses have evolved a way of encapsulating and delivering their genes to human cells in a pathogenic manner. Scientists have tried to harness this ability by manipulating the viral genome to remove disease- causing genes and insert therapeutic ones.

15. VIRUS Virus bind to their hosts and introduce their genetic material into the host cell. Plausible strategy for gene therapy, by removing the viral DNA and using the virus as a vehicle to deliver the therapeutic DNA. The viruses used are altered to make them safe, although some risks still exist with gene therapy.

16. TYPES OF VIRUS Many GT clinical trials rely on retroviruses or adenoviruses to deliver the desired gene. Other viruses used as vectors include adeno-associated viruses, lentiviruses, pox viruses, alphaviruses, and herpes viruses. Differ in how well they transfer genes to the cells they recognize and are able to infect, and whether they alter the cell’s DNA permanently or temporarily

18. VIRAL VECTOR Are a tool commonly used by molecular biologists to deliver genetic material into cells. Can be performed in vivo or in vitro. Viruses have evolved specialized molecular mechanisms to efficiently transport their genomes inside the cells they infect. Delivery of genes by a virus is termed transduction and the infected cells are described as transduced.

21. retrovirus

22. NON VIRAL VECTOR Methods of non-viral gene delivery have also been explored using physical (carrier-free gene delivery) and chemical approaches (synthetic vector-based gene delivery).

23. PHYSICAL METHOD Physical approaches, including  Needle injection  Electroporation  Gene gun  Ultrasound  Hydrodynamic delivery employ a physical force that permeates the cell membrane and facilitates intracellular gene transfer

24. I.NAKED DNA The simplest method of non-viral transfection. Clinical trials carried out of intramuscular injection of a naked DNA plasmid have occurred with some success; however, the expression has been very low in comparison to other methods of transfection.

25. This success, however, does not compare to that of the other methods, leading to research into more efficient methods for delivery of the naked DNA such as electroporation and the use of a "gene gun", which shoots DNA coated gold particles into the cell using high pressure gas.

26. Gene gun

27. CHEMICAL METHODS THAT ENHANCE THE DELIVERY OF GENE THERAPY -liposome

29. DNA must be protected from damage & its entry into the cell must be facilitated Plasmid DNA can be covered with lipids in an organized structure

30. Common used of lipoplexes In gene transfer into cancer cells, where the supplied genes have activated tumor suppressor control genes in the cell decrease the activity of oncogenes. useful in transfecting respiratory epithelial cells, so they may be used for treatment of genetic respiratory diseases such as cystic fibrosis.

33. Give a chance of a normal life to baby born with genetic disease. Give hope of healthy life to cancer patient. For certain disease that do not have any cure except gene therapy, it could save many lives ADVANTAGES OF GENE THERAPY

34. The genetic testing, screening and research in finding the availability of certain gene is very controversy. May increase rate of abortion if prenatal test regarding baby with genetic disease is done. The cost is very high and the patient might need an insurance to cover the treatment. Cosmetic industry may monopolized this gene therapy if it is used in enhancing beauty and in vanishing the aging effect, rather than used for treatment of a disease. DISADVANTAGES OF GENE THERAPY

35. ETHICAL QUESTIONS SURROUNDING GENE THERAPY How can “good” and “bad” uses of gene therapy be distinguished? Who decides which traits are normal and which constitute a disability or disorder? Will the therapy only benefit the wealthy due to its high cost? Could the widespread use of gene therapy make the society less accepting 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?

36. DR.AMINA EL SHAIBANY PHG424 2014