Hypoxic and pharmacological activation of HIF inhibits SARS-CoV-2 infection of lung epithelial cells.

Peter Ac Wing ,Maria Prange-Barczynska,Peter J Ratcliffe,Tammie Bishop,Marko Noerenberg,Isobel L.A Argles,Ilan Davis,Samvid Kurlekar,Senko Tsukuda,Emma J Hodson,Xiaodong Zhuang,William James,Thomas P Keeley,Jane A Mckeating,Peter Balfe,Koichi Watashi,Young Seok Lee ,Craig Thompson ,Sophie Morgan ,Adam Harding ,Kuan Ying A Huang ,Alfredo Castelló ,Timothy S C Hinks

doi:10.1016/j.celrep.2021.109020

Abstract

SummaryCOVID-19, caused by the novel coronavirus SARS-CoV-2, is a global health issue with more than 2 million fatalities to date. Viral replication is shaped by the cellular microenvironment, and one important factor to consider is oxygen tension, in which hypoxia inducible factor (HIF) regulates transcriptional responses to hypoxia. SARS-CoV-2 primarily infects cells of the respiratory tract, entering via its spike glycoprotein binding to angiotensin-converting enzyme 2 (ACE2). We demonstrate that hypoxia and the HIF prolyl hydroxylase inhibitor Roxadustat reduce ACE2 expression and inhibit SARS-CoV-2 entry and replication in lung epithelial cells via an HIF-1α-dependent pathway. Hypoxia and Roxadustat inhibit SARS-CoV-2 RNA replication, showing that post-entry steps in the viral life cycle are oxygen sensitive. This study highlights the importance of HIF signaling in regulating multiple aspects of SARS-CoV-2 infection and raises the potential use of HIF prolyl hydroxylase inhibitors in the prevention or treatment of COVID-19.

Highlights

(TMPRSS2) triggers fusion of the viral and cell membranes (Hoffmann et al, 2020; Wan et al, 2020)
Because the effects of low oxygen on both angiotensin-converting enzyme 2 (ACE2) expression and SARS-CoV-2 replication are likely to be cell context dependent, we evaluated whether hypoxia alters SARS-CoV-2 entry and replication in lung epithelial cells
To define whether hypoxia altered the infectivity of SARS-CoV-2 particles, we assessed the ratio of RNA copies per plaque-forming unit (PFU), finding no significant difference between virus produced from cells at either 18% O2 or 1% O2

Summary

SUMMARY

COVID-19, caused by the novel coronavirus SARS-CoV-2, is a global health issue with more than 2 million fatalities to date. SiRNA silencing of HIF-1a, but not HIF-2a, in Calu-3 cells reversed the hypoxic or FG-4592-mediated suppression of viral infection, demonstrating a role for HIF-1a in repressing SARS-CoV-2 RNA replication (Figure 2H) These data show a key role for HIF-1a in repressing ACE2-dependent, authentic SARS-CoV-2 entry and infection. A key finding from our study is the potential therapeutic application of Roxadustat, and other related HIF prolyl hydroxylase inhibitors, in COVID-19, especially because these act on multiple stages of the viral life cycle (impairing entry and replication) and, as such, may be effective against emerging SARS-CoV-2 variants These drugs have been developed as erythropoiesis-stimulating agents in patients with anemic and chronic kidney disease and are currently being used in both pre-dialysis and dialysis settings. B Lead contact B Materials availability B Data and code availability d EXPERIMENTAL MODEL AND SUBJECT DETAILS B Animals B Cell culture B Viral strains d METHOD DETAILS B SARS-CoV-2 pseudoparticle genesis and infection B SARS-CoV-2 propagation and infection B Immunoblotting B RT-qPCR B FISH quantification of SARS-CoV-2 RNA B Image acquisition and analysis B Materials d QUANTIFICATION AND STATISTICAL ANALYSIS

DECLARATION OF INTERESTS

Findings

METHOD DETAILS