Novel strategy for treating neurotropic viral infections using hypolipidemic drug Atorvastatin

Abstract

18Chandipura virus (CHPV) and Japanese Encephalitis Virus (JEV) are known to infect neurons 19 followed by their successful propagation. Increased incidences of central nervous system 20 invasion by the abovementioned viruses have been reported in case of children and elderly thus 21 culminating into severe neurological damage. Literature suggests induction of endoplasmic 22 reticulum (ER)-stress related proteins upon CHPV and JEV infection which help promote viral 23 reproduction. Since earlier studies underscore the pleotropic role of atorvastatin (AT) in 24 neuroprotection against flaviviruses like Hepatitis C and dengue, it was hypothesized that AT 25 might also act as a neuroprotective agent against RNA viruses like CHPV and JEV. AT-26 mediated antiviral activity was evaluated by assessing survivability of virus-infected mouse pups 27 treated with the drug. Balb C mice were used for in vivo experiments. Neuro2A cell line was 28 used as the model for in vitro experiments. Cells subjected to AT treatment were infected by 29 CHPV and JEV followed by evaluation of ER stress-related and apoptosis-related proteins by 30 immunoblotting technique and immunofluorescence microscopy. Interaction of host protein with 31 viral genome was assessed by RNA-Co-IP. AT treatment exhibited significant anti-viral activity 32 against CHPV and JEV infections via hnRNPC-dependent manner. Viral genome-hnRNPC 33 interaction was found to be abrogated upon AT action. AT was also observed to reduce secretion 34 of proinflammatory cytokines by the neurons in response to viral infection. Moreover, AT 35 treatment was also demonstrated to reduce neuronal death by abrogating virus-induced miR-21 36 upregulation in hnRNPC-dependent fashion. This study thus suggests probable candidature of 37 AT as antiviral against CHPV and JEV infections.38 Introduction: 39 Incidences of Acute Encephalitis Syndrome (AES) resulting from infection by neurotropic RNA 40 viruses like Japanese Encephalitis Virus (JEV) and Chandipura Virus (CHPV) have been 41 reported in India on an annual basis (1). Both JEV and CHPV are arboviruses differing in terms 42 of genetic material. JEV possesses a single positive-stranded RNA as genetic material, while 43 CHPV is a negative-stranded RNA virus (2, 3). JEV is being reported to mainly target central 44 nervous system (CNS) of children. Since CNS of infants undergo a lot of physiological changes 45 as development progresses, any insult to neuronal physiology may lead to severe perturbation of 46 neurocognitive abilities thus leaving the survivors of JEV-induced encephalitis with motor 47 deficits, cognitive and language impairments and learning difficulties (4, 5). Whereas, although 48 infection due to CHPV leads to acute encephalopathy, no report till date documents incidences of 49 neurological sequel experienced by survivors of CHPV-induced encephalitis (5). Evidences 50 suggest neuronal death following JEV or CHPV infections by two distinct mechanisms. In 51 addition to direct viral propagation-mediated neuronal res...