5.

Gene Therapy GENE THERAPY The replacement of faulty genes with healthy genes to cure or prevent conditions from genetic abnormalities -> Somatic Gene Therapy - insert -> appropriate cells of diseased - change NOT passed onto offspring -> Germ line Therapy - insert -> gamete - change PASSED onto offspring In-vivo + Ex-vivo Strategies: -> Ex-vivo - normal gene removed from cell - transferred to target cells in culture medium (in vitro) -> multiply + reproduce - harvested -> In-vivo Normal genes transferred inside body via sprays Vector: Recombinant retrovirus adenovirus herpes simplex virus 1. disease-causing gene removed 2. healthy recombinant gene needed to correct disorder inserted 3. Cloned + injected

Viral vector effective as viruses are capable for introducing genes into human cells, since they naturally reproduce by delivering their genetic info into our cells. Problem: Patients immune system destroys viral vector detected as foreign virus ADA therapy worked well since it used blood ex-vivo (an approach not possible for most other disease) Other Vectors: -> Naked DNA directly injected into cells; attached to gold particles + fired into tissue -> Liposomes tiny balls lipid filled with DNA

CURRENT USE X-SCID (ADA deficieny) Cause: unable to produce enzyme adenosine deaminase -> buildup of adenosine -> dmg lymphocyte Symptoms: prone to infections, lymphomas (cancer of lymphocytes) Treatment: retrovirus with normal ADA gene -> insert ex-vivo -> removed + cultured stem cells corrected cells cloned -> injected back into body Result: Healthier, antibody levels increase, T cell no increase Cystic Fibrosis Cause: mutated gene -> defective protein -> dehydrates mucus membrane -> thick sticky mucus Symptoms: mucus cant be cleared -> improper respiratory functioning + prone to infection, lung disease Treatment: good CF gene -> Adenovirus (affect respiratory tissue) -> nose drop, spray, drips -> lung cells -> Liposomes Problem: - no vector that can deliver corrected genes to enough cells to make significant difference - no strong vector that can get in under the radar of the bodys immune system Wilson - CF too large to fit into vector - cells lining respiratory tract die + replaced every few months (if stem cells that produce these cells are identified -> gene therapy -> permanent cure) Current: + Split gene -> 2 parts -> 2 separate AAV (adeno-assoc virus) vector -> aerosol = recombine in patients cells + hybrid vector: HIV virus + decorated with proteins from Ebola virus (BUT not yet found way to use this method w/o risking lethal infection) Cancer -> Replacement Gene Therapy - several types, main focus on cancer-suppressor genes Gene p53 produce protein that shut down cells with DNA dmg est >50% cancers involve p53 gene mutation 1997 Texas Uni researchers - adenovirus -> deliver corrected p53 gene -> 21 adv lung cancer patients (inject directly into cancer) - in most, tumors regressed, esp + anti-cancer drug -> Suicide Gene Therapy (Selective Destruction) - suicide gene -> retrovirus -> cancer cell = produce substance toxic to cancer cells Texas researchers - gene forces cell to produce = enzyme thymidine kinase -> converts drug ganciclovir -> toxic substances - drug ganciclovir -> patient = cancer cells with suicide gene die (selective) = failed most patients BUT very successful where effective (cancers reduced >50%, many as 95% cancer cells died)

Aids - Aus consortium given grant by US NIAID to develop new vaccine for HIV/AIDS - Led by David Cooper - several vaccine trialled, none so far proved effective -> virus keeps changing protein coat + hides in cells where antibodies cannot reach Two-Stage Approach: -> Priming with a naked DNA vaccine made from HIV genes = triggers immune system to produce killer T cells to destroy HIV virus -> Boosting with genetically engineered foulpox virus (vector for HIV gene) = increases strength of T-cell response Foulpox virus vector for HIV gene produces HIV antigens in cytoplasm = trigger antibody + T cell response Animal trials = effective way to induce T cells to destroy HIV-infected cells Human Clinical trials 2003