A key factor in the success of gene therapy is the development of delivery systems that arc capable of efficient gene transfer in a variety of tissues without causing any pathogenic effects. This thesis is aimed at developing gene therapy strategies for the treatment of neurodegenerativc diseases, mainly Spinal Muscular Atrophy (SMA). Integration-deficient lentiviral vectors (lDLVs),. based on HIV -1 through the use of class I integrase mutation, fail to integrate as proviruses and are instead converted into episomal circles, which are metabolically stable. These vectors are of particular use as they support expression of transgenes in quiescent cells and exhibit a decreased risk of insertions I mutagenesis. I have embarked on an extensive study to compare the transduction efficiency of IDLVs and matched integrating lentivectors in spinal cord tissues in vitro and in vivo. My results demonstrate similar efficiency of eGFP expression from both vector forms in most cell types analysed, including motor neurons, inter-neurons and astroglia. After intraspinal injection of IDLY expressing eGFP into adult rat and mouse lumbar cord in vivo, transduction was mainly neuronal, with both motor and inter-neurons being efficiently targeted. Intraspinal injection of mouse embryos revealed that IDLY can completely transduce motor neurons. Additionally, IDLYs produced strong expression of mouse survival motor neuron gene (mSmn; SMA determining gene) in the injected cords. My data also revealed that IDLV-mediated express ion of a growth factor or tetanus toxin fragment C (TTC) rescued motor neuron cultures from death caused by removal of exogenous trophic support. Moreover, integration proficient lentiviral vectors encoding for mSmn restored the number of gems in type I SMA fibroblasts. Taken together, the results of this study strongly implicate that TDLVs could be efficient and safe tools for cord transduction in therapeutic strategies such as for treatment of SMA.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:604282 |
| Date | January 2011 |
| Creators | Ahmed, Sherif |
| Publisher | Royal Holloway, University of London |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
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