HSV-1 vectors are attractive for gene therapy of aging disorders that affect the brain because HSV-1 can persist indefinitely in neurons in the latent state and large HSV-1 vectors can coexpress multiple genes. This laboratory has developed a Herpes Simplex Virus (HSV-1) plasmid vector system for gene transfer into neurons. Using this system, we have begun to explore gene therapy approaches to specific aging disorders that affect the brain, such as Parkinson's Disease (PD). We have shown that delivery of a HSV-1 vector that expresses human tyrosine hydroxylase into the partially denervated striatum in the 6-hydroxydopamine rat model of PD results in long-term (1 year) biochemical and behavioral correction. Other investigators have demonstrated the potential of using this vector system for gene therapy for a number of other neurological disorders. We developed a helper virus-free packaging system for these HSV-1 vectors. This improvement substantially reduces the cytopathic effects and the inflammatory response previously associated with gene transfer. Furthermore, we and others have recently identified specific promoters that support long-term expression in rat forebrain neurons. However, the relatively low titers remain one of the primary barriers to the use of this vector system for human gene therapy of aging disorders that affect the brain. The goal of this proposal is to develop a packaging cell line that can produce high-titer helper virus-free HSV-1 vector stocks. High-titer retrovirus, lentivirus, and adenovirus vector stocks have been produced using packaging cell lines, and these vector stocks have been used in human gene therapy. The first specific aim will isolate a cell line that stably maintains an HSV-1 genome that does not express any immediate early (IE) genes and lacks a packaging site. The second specific aim will produce high-titer, helper virus-free HSV-1 vector stocks by using an HSV-1 vector that contains the 3 essential IE genes flanked by lox sites. This packaging system is based upon standard genetic complementation. High-titer vector stocks will be produced by serial passaging. To excise the IE genes from the vector, the final passage will use a cell line that expresses Cre recombinase. The third specific aim will evaluate the safety features of this packaging system. The fourth specific aim will use these high-titer vector stocks to achieve gene transfer to large numbers of cells in the rat striatum.

National Institute of Health (NIH)
National Institute on Aging (NIA)
Research Project (R01)
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Special Emphasis Panel (ZRG1-BDCN-2 (02))
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Wise, Bradley C
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Harvard University
Schools of Medicine
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Cao, Haiyan; Zhang, Guo-rong; Geller, Alfred I (2011) Antibody-mediated targeted gene transfer of helper virus-free HSV-1 vectors to rat neocortical neurons that contain either NMDA receptor 2B or 2A subunits. Brain Res 1415:127-35
Cao, Haiyan; Zhang, Guo-Rong; Geller, Alfred I (2010) Antibody-mediated targeted gene transfer to NMDA NR1-containing neurons in rat neocortex by helper virus-free HSV-1 vector particles containing a chimeric HSV-1 glycoprotein C-staphylococcus A protein. Brain Res 1351:1-12
Liu, Meng; Wang, Xiaodan; Geller, Alfred I (2009) Improved long-term expression from helper virus-free HSV-1 vectors packaged using combinations of mutated HSV-1 proteins that include the UL13 protein kinase and specific components of the VP16 transcriptional complex. BMC Mol Biol 10:58
Zhang, Guo-Rong; Liu, Meng; Cao, Haiyan et al. (2009) Improved spatial learning in aged rats by genetic activation of protein kinase C in small groups of hippocampal neurons. Hippocampus 19:413-23
Cao, Haiyan; Zhang, Guo-rong; Wang, Xiaodan et al. (2008) Enhanced nigrostriatal neuron-specific, long-term expression by using neural-specific promoters in combination with targeted gene transfer by modified helper virus-free HSV-1 vector particles. BMC Neurosci 9:37
Rasmussen, Morten; Kong, Lingxin; Zhang, Guo-rong et al. (2007) Glutamatergic or GABAergic neuron-specific, long-term expression in neocortical neurons from helper virus-free HSV-1 vectors containing the phosphate-activated glutaminase, vesicular glutamate transporter-1, or glutamic acid decarboxylase promoter. Brain Res 1144:19-32
Gao, Qingshen; Sun, Mei; Wang, Xiaodan et al. (2007) Isolation of an enhancer from the rat tyrosine hydroxylase promoter that supports long-term, neuronal-specific expression from a neurofilament promoter, in a helper virus-free HSV-1 vector system. Brain Res 1130:1-16
Gao, Qingshen; Sun, Mei; Wang, Xiaodan et al. (2006) Long-term inducible expression in striatal neurons from helper virus-free HSV-1 vectors that contain the tetracycline-inducible promoter system. Brain Res 1083:1-13
Wang, Xiaodan; Kong, Lingxin; Zhang, Guo-rong et al. (2005) Targeted gene transfer to nigrostriatal neurons in the rat brain by helper virus-free HSV-1 vector particles that contain either a chimeric HSV-1 glycoprotein C-GDNF or a gC-BDNF protein. Brain Res Mol Brain Res 139:88-102
Sun, Mei; Kong, Lingxin; Wang, Xiaodan et al. (2005) Comparison of the capability of GDNF, BDNF, or both, to protect nigrostriatal neurons in a rat model of Parkinson's disease. Brain Res 1052:119-29

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