Cd302 and Cr1l restrict human hepatotropic virus cross-species transmission to mice [6PMH siRNA]

Virus species- and tissue-tropism is governed by host dependency and restriction factors. Hepatitis C virus (HCV) exhibits a narrow species-tropism and murine hepatocytes are refractory to infection. Using murine liver cDNA library screening we identified Cd302, a lectin, and Cr1l, a complement receptor, as pan-genotypic restrictors of HCV infection. Cd302/Cr1l interact to impede virion uptake and co-operatively induce a non-canonical transcriptional program, inhibiting HCV and hepatitis B virus (HBV) infection in vitro. CAS9 disruption of murine hepatocyte Cd302 expression increased HCV permissiveness in-vivo and ex-vivo, and modulated the intrinsic hepatocyte transcriptome dysregulating metabolic process and host defense genes. In contrast, co-operative CD302/CR1L expression was absent and HCV restriction reduced in human hepatocytes. The Cd302/Cr1l axis therefore contributes to limiting hepatotropic virus cross-species transmission to mice, opening new avenues for step-wise development of mouse models for these important human pathogens, which cause substantial disease burden globally. Overall design: Primary mouse hepatocytes from x3 entry receptor transgenic mice with conditional liver expression (human Occludin and CD81: hOChep) and one wildtype control mouse were isolated after liver perfusion and liberase digestion. Hepatoyctes were plated onto 6-well tissue culture dishes dishes and cultured in HCM. 24 hours after plating hepatocytes were transfected with siRNAs targeting the genes of interest or a control siRNA. 24 hours after siRNA transfection hepatocytes were infected with HCV (strain Jc1, MOI 1) or left uninfected. 24 hours post infection RNA was isolated for RNA-seq.