2. Gene Therapy Mostafa A. Askar NCRRT By M.Sc. In Molecular Biology
Molecular Immunology Specialist
NCRRT

3. Lecture Content Definition of Gene Therapy Aim of Gene Therapy
Types of Gene Therapy
General Principles of Gene Therapy
DNA delivery in Gene Therapy; How & With What
Gene Therapy Approach
Gene Therapy Problems
Inherited Diseases Treated by Gene Therapy; Trails

4. Lecture Content Definition of Gene Therapy Aim of Gene Therapy
Types of Gene Therapy
General Principles of Gene Therapy
DNA delivery in Gene Therapy; How & With What
Gene Therapy Approach
Gene Therapy Problems
Inherited Diseases Treated by Gene Therapy; Trails

5. Definition of Gene Therapy
Gene Therapy: is the insertion, alteration, or removal of genes within an individual's cells and biological tissues to treat disease.
It is a technique for correcting defective genes.

6. Lecture Content Definition of Gene Therapy Aim of Gene Therapy
Types of Gene Therapy
General Principles of Gene Therapy
DNA delivery in Gene Therapy; How & With What
Gene Therapy Approach
Gene Therapy Problems
Inherited Diseases Treated by Gene Therapy; Trails

7. Aim of Gene Therapy
The most common form of gene therapy involves the insertion of functional genes into an unspecified genomic location in order to replace a mutated gene, but other forms involve directly correcting the mutation or modifying normal gene that
enables a viral infection.

8. Lecture Content Definition of Gene Therapy Aim of Gene Therapy
Types of Gene Therapy
General Principles of Gene Therapy
DNA delivery in Gene Therapy; How & With What
Gene Therapy Approach
Gene Therapy Problems
Inherited Diseases Treated by Gene Therapy; Trails

9. Somatic gene therapy In Utero gene therapy Germ line gene therapy
Types of gene therapy
Somatic gene therapy
In Utero gene therapy
Germ line gene therapy

10. Somatic gene therapy
In the case of somatic gene therapy, the therapeutic genes are transferred into the somatic cells of a patient. Any modifications and effects will be restricted to the individual patient only, and will not be inherited by the patient's offspring or later generations.

11. In Utero gene therapy
It involves gene therapy of fetuses whilst in the womb. There is some concern that the germline
will be affected.

12. Germ line gene therapy
In this case; Germ cells (Ex. sperm or eggs) are modified by the introduction of functional genes, which are integrated into their genomes. Therefore, the change due to therapy would be heritable and would be passed on to later generations.

13. Lecture Content Definition of Gene Therapy Aim of Gene Therapy
Types of Gene Therapy
General Principles of Gene Therapy
DNA delivery in Gene Therapy; How & With What
Gene Therapy Approach
Gene Therapy Problems
Inherited Diseases Treated by Gene Therapy; Trails

14. General principles of Gene therapy
Identification and characterization of gene
Cloning of gene
Choice of vector
Method of delivery
Expression of gene

15. Characteristics of the ideal vector
Non-pathogeneic.
Capable of integrating DNA into host genome at a single site.
Able to carry large amounts of DNA.
Unable to cause immune response.
Target cells specifically.

16. Lecture Content Definition of Gene Therapy Aim of Gene Therapy
Types of Gene Therapy
General Principles of Gene Therapy
DNA delivery in Gene Therapy; How & With What
Gene Therapy Approach
Gene Therapy Problems
Inherited Diseases Treated by Gene Therapy; Trails

17. Viral vectors. Non-viral delivery.
DNA delivery is mainly accomplished by either:
Viral vectors.
Non-viral delivery.

18. Vectors Adenoviruses Adeno Associated viruses Reto viruses
Herpes Simplex viruses

19. Adenoviruses
* dsDNA Viruses. * Non-integrating viruses. * Infect dividing & non-dividing cells.

20. Adenovirus vector in Gene Therapy

21. Adenoviruses Advantages. Non-oncogenic. Relatively easy to culture
Adenoviruses Advantages * Non-oncogenic. * Relatively easy to culture. * Proteins functions are known. * Complete DNA sequence is available.
Adenoviruses Disadvantages
Transient expression leads to multiple injections

22. * RNA Viruses. * Incorporating Viruses. * Infect dividing cells.
Retroviruses
* RNA Viruses. * Incorporating Viruses. * Infect dividing cells.

23. Retroviruses
Advantages
* Their ability to integrate in host DNA.
Disadvantages
* Infect ONLY dividing cells * Integration is random, causing “Insertional Mutgenesis”.

24. Non-viral delivery in gene therapy
Particle bombardment
Receptor-mediated uptake
Liposomes

25. Particle bombardment
Particle bombardment. DNA is fired through the cell walls and membranes on metal particles. This method was originally developed to get DNA into plants. However, it has also been used to make transgenic animals and is occasionally used for humans.

26. Receptor-mediated uptake
Receptor-mediated uptake. DNA is attached to a protein that is recognized by a cell surface receptor. When the protein enters the cell, the DNA is taken in with it.

27. Liposomes
Liposomes are spherical vesicles composed of phospholipid. They have been used in around 10% of gene therapy trials.

28. Lecture Content Definition of Gene Therapy Aim of Gene Therapy
Types of Gene Therapy
General Principles of Gene Therapy
DNA delivery in Gene Therapy; How & With What
Gene Therapy Approach
Gene Therapy Problems
Inherited Diseases Treated by Gene Therapy; Trails

29. Antisense Gene Therapy

30. Gene Therapy to Induce Apoptosis

31. Anti-Angiogenic Gene Therapy

32. Lecture Content Definition of Gene Therapy Aim of Gene Therapy
Types of Gene Therapy
General Principles of Gene Therapy
DNA delivery in Gene Therapy; How & With What
Gene Therapy Approach
Gene Therapy Problems
Inherited Diseases Treated by Gene Therapy; Trails

33. Problems with Gene Therapy
Short Lived
2. Immune Response
3. Viral Vectors
may cause disease once inside

34. Problems with Gene Therapy
4. Multigene Disorders
Heart disease, high blood pressure, Alzheimer’s, arthritis and diabetes are hard to treat because you need to introduce more than one gene
5. May induce a tumor if integrated in a tumor suppressor gene because insertional mutagenesis

35. Lecture Content Definition of Gene Therapy Aim of Gene Therapy
Types of Gene Therapy
General Principles of Gene Therapy
DNA delivery in Gene Therapy; How & With What
Gene Therapy Approach
Gene Therapy Problems
Inherited Diseases Treated by Gene Therapy; Trails

36. Gene Therapy trials for inherited diseases
Lung
Cystic Fibrosis (CF).

37. Diseases Treated by Gene Therapy
Bone Marrow Adenosene Deaminase Deficency(ADA)
Brain (Alzheimer’s disease)
Liver (Factor VIII deficiency.)

38. Questions?