STEREOTAXIC GENE THERAPY TO THE RHESUS CNS

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The central nervous system is a primary target for gene therapy because the broad spectrum of inherited and acquired disorders that affect the brain. Neurodegenerative diseases such as Parkinson¿s disease, Huntington¿s disease, Alzheimer¿s disease, and amyotrophic lateral sclerosis (ALS) are among the most common disorders that affect older humans and as such are prime targets for the development of gene therapy. Although rare, inherited lysosomal storage disorders and leukodystrophys, such as Tay-Sach¿s, mucopolysaccharidosis and Krabbe¿s disease, are also primary targets for the development of novel gene therapy interventions. Treatment options for these and other CNS disorders are primarily limited to supportive care, thus providing few therapeutic options. Direct CNS gene transfer may provide the best opportunity for eliminating the pathology associated with these conditions. The goal of this proposal is the comparison of the transduction of recombinant lentivirus (LV) and adeno-associated virus (AAV) vector systems with regard to levels of gene transfer, pattern of (cells and structures), and persistence of gene expression in the CNS of nonhuman primates, specifically rhesus macaques. The ultimate aim of this proposal is to determine which vector or combination of vectors provides high levels of gene transfer to the broadest range of cell types in an animal model that is very closely related to humans. The primary advantage of this proposal is that preparations of both vector systems will be prepared using consistent production protocols and administered directly to the CNS. We have performed a series of gene transfer and biodstribution studies using direct stereotaxic injection of these recombinant virus preparations to target multiple regions of the brain, such as the hippocampus, cortex, caudate putamen/striatum, thalamus and cerebellum. Following administration of the vector preparations, we are currently mapping the transduction patterns of the various vectors by determining the levels of gene transfer to selected regions of the brain and delineating the specific cell types transduced by immunohistochemistry and confocal microscopy. Our current data suggests that marked differences in the patterns of gene transfer provided by various serotypes of AAV (AAV 1, 2, 7, 9 and 10) vectors can be detected. The biodistribution of VSV-G pseudotyped lentivirus vectors demonstrated broad gene transfer to many cell types, but at lower efficiency than AAV vectors. . The results of the proposed translational studies will serve as invaluable information on the design and execution of CNS-directed gene therapy protocols in humans, especially children.