RT Journal Article
SR Electronic
T1 CRISPRa screening with real world evidence identifies potassium channels as neuronal entry factors and druggable targets for SARS-CoV-2
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.07.01.450475
DO 10.1101/2021.07.01.450475
A1 Chengkun Wang
A1 Ravi K. Dinesh
A1 Yuanhao Qu
A1 Arjun Rustagi
A1 Henry Cousins
A1 James Zengel
A1 Yinglong Guo
A1 Taryn Hall
A1 Aimee Beck
A1 Luke Tso
A1 Elif Tokar Erdemic
A1 Kae Tanudtanud
A1 Sheng Ren
A1 Kathy Tzy-Hwa Tzeng
A1 Aaron Wilk
A1 Mengdi Wang
A1 Jan Carette
A1 Russ Altman
A1 Catherine A. Blish
A1 Le Cong
YR 2021
UL http://biorxiv.org/content/early/2021/07/01/2021.07.01.450475.abstract
AB Although vaccines for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been successful, there are no good treatments for those who are actively infected and potentially suffer from diverse neurological symptoms. While SARS-CoV-2 primarily infects the respiratory tract, clinical evidence indicates that cells from sensory organs, brain, and heart are also susceptible to infection. An understanding of factors critical for viral infection in these tissues may help identify novel therapeutics. To discover host factors involved in SARS-CoV-2 viral entry, we performed CRISPR activation (CRISPRa) screens targeting all 6000+ human membrane proteins in cells with and without overexpression of ACE2 using Spike-pseudotyped lentiviruses. We identified both novel as well as previously validated host factors. Notably, we used replication-competent SARS-CoV-2 to validate new viral-entry promoting genes, including potassium channel KCNA6, protease LGMN, and MHC-II component HLA-DPB1. We found that the overexpression of KCNA6 led to a marked increase in infection even in cells with undetectable levels of ACE2 expression. Our analysis of human olfactory neuroepithelium scRNA-seq data revealed that OLIG2+ cells--previously identified as sites of infection in COVID-19 autopsy studies--have high KCNA6 expression and minimal levels of ACE2, suggesting that the presence of KCNA6 may explain sensory/neuronal aspects of COVID-19 symptoms. Further, we demonstrate that FDA-approved compound dalfampridine, an inhibitor of KCNA-family potassium channels, suppresses viral entry in a dosage-dependent manner. Finally, we identified common prescription drugs likely to modulate the top screen hits. We then performed a retrospective analysis of insurance claims of ∼8 million patients and found a clinical association between screen-identified drug classes, particularly those targeting potassium channels, and COVID-19 severity. We have thus identified the potassium channel KCNA6 as a SARS-CoV-2 host factor, expanded our understanding of potential viral tropism, and identified promising targets for drug repurposing and development.Competing Interest StatementStanford University has filed patent applications based on this work. The authors will make all reagents openly available to the academic community.