Genome-wide screening of host genes involved in virus-induced neurotoxic signaling using lentiviral siRNA library

Tsuneya Ikezu, MD, PhD
Clinton Jones, PhD

Viral infection-mediated neurotoxicity is a central pathologic mechanism among epidemic viral encephalitis, including human immunodeficiency virus (HIV)-1 and herpes simplex virus (HSV) encephalitis.  Survival of neurons chronic viral infection is thus important for maintaining the cognitive, motor, and psychiatric function of brain.  Identification of host genes responsible for the neurotoxic signaling will enable us to enhance the cell survival and enable us to develop a therapeutic invention.  To this end, we will try high-throughput genome-wide screening of host genes, which are responsible for the viral neurotoxic signaling in human neuronal cell line.  Lentiviral siRNA library targeting 47,400 human mRNA transcripts will utilized to identify siRNA clones which can rescue cell death by HIV-1 gp120, HSV, or HSV LAT null mutant virus infection.  The siRNA clones will be identified by Affymetrix microarrary system, which has corresponding spots for siRNA oligo sequence for rapid identification of candidate siRNA clons.  The identified siRNA clones by the library screening will be further tested individually both in vitro and in vivo using virus-infected anima models.  This study is a saturated screening of all human genes, high throughput, and can be easily confirmed both in vitro and in vivo using psedudotyped lentivirus vector system.  Such genome-wide screening of host genes using siRNA lentivirus has never been attempted.  The data obtained from the study will be readily available for future funding, since this is a genome-wide functional screening of biologically significant neurotoxic signaling, and has a therapeutic potential.