GENOME-WIDE SCREENING OF HOST GENES INVOLVED IN VIRUS-INDUCED NEUROTOXIC SIGNALI

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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. Viral infection-mediated neurotoxicity is a central pathologic mechanism for epidemic viral encephalitis, including HAD (human immunodeficiency virus (HIV)-1 associated dementia) and herpes simplex virus (HSV) encephalitis. Survival of neurons during chronic viral infection is important for maintaining the cognitive, motor, and psychiatric functions of the brain. Identification of the host genes responsible for neurotoxic signaling will enable us to enhance cell survival and to develop a therapeutic intervention. To identify which genes are responsible for viral neurotoxic signaling, we will utilize high-throughput genome-wide screening of host genes in a human neuronal cell line. A lentiviral siRNA library that targets 47,400 human mRNA transcripts will be utilized to identify which siRNA clones can prevent in vitro human neuronal cell death induced by incubation with HIV-1 gp120, or by infection with HSV, or HSV LAT null mutant virus. The siRNA clones will be identified by GeneChip microarray system, which has all the corresponding spots for siRNA oligo sequence for rapid identification of candidate siRNA clones. Each siRNA clone identified by the library screening will be further tested for their neuroprotective effect both in vitro and in vivo using virus-infected animal models. This study is a high throughput saturated screening of all human genes with the results being easily confirmed both in vitro and in vivo using a pseudotyped lentivirus vector system. The data obtained from the study can be readily used in support of future funding, since this is a genome-wide functional screening of biologically significant neurotoxic signaling with therapeutic potential.