Abstract
Better understanding on interactions between SARS-CoV-2 and host cells should help to identify host factors that may be targetable to combat infection and COVID-19 pathology. To this end, we have conducted a genome-wide CRISPR/Cas9-based loss-of-function screen in human lung cancer cells infected with SARS-CoV-2-pseudotyped lentiviruses. Our results recapitulate many findings from previous screens that used full SARS-CoV-2 viruses, but also unveil two novel critical host factors: the lysosomal efflux transporter SPNS1 and the plasma and lysosomal membrane protein PLAC8. Functional experiments with full SARS-CoV-2 viruses confirm that loss-of-function of these genes impairs viral entry. We find that PLAC8 is a key limiting host factor, whose overexpression boosts viral infection in eight different human lung cancer cell lines. Using single-cell RNA-Seq data analyses, we demonstrate that PLAC8 is highly expressed in ciliated and secretory cells of the respiratory tract, as well as in gut enterocytes, cell types that are highly susceptible to SARS-CoV-2 infection. Proteomics and cell biology studies suggest that PLAC8 and SPNS1 regulate the autophagolysosomal compartment and affect the intracellular fate of endocytosed virions.
Keywords: autophagy; covid19; genetic screen; plac8; spns1.
【저자키워드】 COVID19, autophagy, genetic screen, spns1., plac8, 【초록키워드】 SARS-CoV-2, viral infection, SARS-COV-2 infection, Genetic, Infection, viral entry, Protein, human lung, plasma, respiratory tract, experiment, Critical, single-cell, boost, Interaction, regulate, Gut, enterocytes, cell type, host cell, Factor, help, SARS-CoV-2 viruses, loss-of-function, COVID-19 pathology, overexpression, virions, lentiviruses, Host, Affect, susceptible, Cell, cancer cell lines, identify, conducted, eight, expressed, ciliated, analyses, Better, impair, lysosomal, cancer cell, endocytosed, lysosomal membrane, SARS-CoV-2 virus, secretory cell, SPNS1, 【제목키워드】 Protein, plasma, SARS-CoV-2 entry, human cell, Host, lysosomal membrane,