RNA interference based membrane trafficking screen identifies drug targets for Japanese Encephalitis Virus
Renu Khasa1,2 Suvadip Mallick3, Anirban Basu3, Sudhanshu Vrati4 and Manjula Kalia1
1Translational Health Science and Technology Institute, Faridabad, Haryana, India; 2Symbiosis International University (SIU), Pune, Maharashtra, India; 3National Brain Research Centre, Gurugram, Haryana, India; 4Regional Centre for Biotechnology, Faridabad, Haryana, India
Japanese encephalitis virus (JEV), belongs to the family Flaviviridae and is spread by mosquito bites. The disease is a major public health concern for India with potential to become a global pathogen. To identify crucial host proteins that are exploited by the virus to gain entry and establish infection in permissive mammalian cells, we have performed an RNA interference based screen of human genes implicated in membrane trafficking. A panel of 150 membrane trafficking genes were tested in epithelial (HeLa) and neuronal (IMR32) cells for the effect of their depletion on JEV entry, replication and production of infectious virus particles.
Of the several hits that were obtained, eight genes/proteins were identified that are established drug targets, and thus have the potential to be developed as anti-virals for JEV. These drugs are currently being tested in vitro for their inhibitory effect on virus replication. Two drugs- NMS-583 and CB-5083 that are established inhibitors of Valosin-containing protein (VCP) showed attenuation of JEV replication in vitro, and also conferred protection against JEV-induced mortality in the mouse model.
This work is supported by DBT grant BT/MB/01/VIDRC/08 and THSTI intra-mural funds to MK and SV. SM and AB are supported by Tata Innovation Fellowship from the Department of Biotechnology, Government of India (BT/HRD/35/01/02/2014). RK is supported by ICMR-SRF fellowship