Researchers uncover the host protease CAPN2 as a novel host factor that aids the infection of SARS-CoV-2 biorxivpreprint WUSTLmed SARSCoV2 COVID19 coronavirus covid infection
By Pooja Toshniwal PahariaDec 5 2022Reviewed by Danielle Ellis, B.Sc. In a recent study posted to the bioRxiv* preprint server, researchers investigated the potential of the calpain-2 molecule as a therapeutic target against .
The authors had recently screened for antiviral compounds using Vero E6 cells and recombinant SARS-CoV-2mNeonGreen virus, wherein several compounds demonstrated anti-SARS-CoV-2 efficacy, and the top 18 hits were validated in the present study. For the validation experiments, recombinant vesicular stomatitis virus e-green fluorescent protein reporter viruses encoding either native VSV-G or the spike protein of SARS-CoV-2 were used.
Further, western blot analysis was performed to validate CAPN2 KO efficiency. CAPN2 KO and wild-type cells were inoculated with SARS-CoV-2 S protein expressing chimeric VSV . Genetics & Genomics eBook Compilation of the top interviews, articles, and news in the last year. Download a free copy Co-transfection experiments were performed using ACE2-expressing human embryonic kidney 293 cells and WA1 strain S protein, with CAPN2, furin, or TMPRSS2 , key proteases reported for cleaving the S protein for efficient host invasion.
IMBX , Nigericin, and brefeldin A showed EC50 values below 2.0 μM against infections of VSV and VSV-SARS-CoV-2. Nitazoxanide showed SARS-CoV-2 inhibition, and MG132 was 100-fold selective in activity against VSV and VSV-SARS-CoV-2 with EC50 values of 44 and 0.6 µM, respectively. No inhibitor except calpeptin showed cytotoxicity. VSV-SARS-CoV-2 mRNA was lowered by 4.0-fold in cells lacking CAPN2. VSV-SARS-CoV-2 plaques were 1.0 mm and 2.0mm in size in KO and WT cells, respectively.
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