RNAi screening using HK-2 cells: insights into the roles of soluble ACE2 in SARS-CoV-2 infection. Yeung et al
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can cause acute respiratory disease and multiorgan failure. Finding human host factors that are essential for SARS-CoV-2 infection could facilitate the formulation of treatment strategies. Using a human kidney cell line—HK-2—that is highly susceptible to SARS-CoV-2, we performed a genome-wide RNAi screening and identified virus dependency factors (VDFs), which play regulatory roles in biological pathways linked to clinical manifestations of SARS-CoV-2 infection. Importantly, we discovered an overlooked role of a secretory form of SARS-CoV-2 receptor, soluble angiotensin converting enzyme 2 (sACE2), in SARS-CoV-2 infection. Further investigations revealed that SARS-CoV-2 exploits receptor-mediated endocytosis through interaction between its spike with sACE2 or sACE2-vasopressin via AT1 or AVPR1B, respectively. Our identification of new VDFs and the regulatory effect of sACE2 on SARS-CoV-2 infection shed new insights into pathogenesis and cell entry mechanism of SARS-CoV-2 as well as potential treatment strategies for COVID-19.
Steps to reproduce
shRNA counts were median-normalized and analyzed with DESeq2 (Love et al., 2014) to infer changes between samples and to evaluate statistical significance. Next, we adjusted the gene counts of raw reads within each treatment group by median normalization. For each treatment group, the gene counts with missing values from individual replicates were removed. The resulting datasets were subjected to clustering analyses, including principal components analysis (PCA), volcano plot, and heat map hierarchical clustering analysis, using the stat function in R, ggplot2 function in R, and Complex Heatmap, respectively (Gu et al., 2016; Ito and Murphy, 2013).