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

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 病毒感染介导的神经毒性是流行性病毒性脑炎（包括HAD（人类免疫缺陷病毒（HIV）-1相关性痴呆）和单纯疱疹病毒（HSV）脑炎）的主要病理机制。 慢性病毒感染期间神经元的存活对于维持大脑的认知、运动和精神功能是重要的。 鉴定负责神经毒性信号传导的宿主基因将使我们能够提高细胞存活率并开发治疗干预。 为了确定哪些基因负责病毒神经毒性信号，我们将利用高通量全基因组筛选人类神经元细胞系中的宿主基因。 将利用靶向47，400个人mRNA转录物的慢病毒siRNA文库来鉴定哪些siRNA克隆可以预防由与HIV-1 gp 120孵育或由HSV或HSV LAT无效突变病毒感染诱导的体外人神经元细胞死亡。 将通过基因芯片微阵列系统鉴定siRNA克隆，该系统具有siRNA寡核苷酸序列的所有对应斑点，用于快速鉴定候选siRNA克隆。 通过文库筛选鉴定的每个siRNA克隆将使用病毒感染的动物模型在体外和体内进一步测试其神经保护作用。 这项研究是一个高通量的饱和筛选的所有人类基因的结果很容易证实在体外和体内使用假型慢病毒载体系统。从研究中获得的数据可以很容易地用于支持未来的资金，因为这是一个具有治疗潜力的生物学意义的神经毒性信号的全基因组功能筛选。

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.