Dependence of SARS-CoV-2 infection on cholesterol-rich lipid raft and endosomal acidification

Xiaowei Li; Wenhua Zhu; Meiyang Fan; Jing Zhang; Yizhao Peng; Fumeng Huang; Nan Wang; Langchong He; Lei Zhang; Rikard Holmdahl; Liesu Meng; Shemin Lu

doi:10.1016/j.csbj.2021.04.001

Dependence of SARS-CoV-2 infection on cholesterol-rich lipid raft and endosomal acidification

Comput Struct Biotechnol J. 2021:19:1933-1943. doi: 10.1016/j.csbj.2021.04.001. Epub 2021 Apr 8.

Authors

Xiaowei Li^{1

2

3}, Wenhua Zhu^{2

3}, Meiyang Fan^{2

3}, Jing Zhang^{2

3}, Yizhao Peng^{2

3}, Fumeng Huang^{2

3}, Nan Wang⁴, Langchong He⁴, Lei Zhang⁵, Rikard Holmdahl^{1

2

3

6}, Liesu Meng^{1

2

3}, Shemin Lu^{1

2

3}

Affiliations

¹ National Joint Engineering Research Center of Biodiagnostics and Biotherapy, Second Affiliated Hospital, Xi'an Jiaotong University, 710004 Xi'an, China.
² Institute of Molecular and Translational Medicine (IMTM), and Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, 710061 Xi'an, China.
³ Key Laboratory of Environment and Genes Related to Diseases (Xi'an Jiaotong University), Ministry of Education of China, 710061 Xi'an, China.
⁴ School of Pharmacy, Xi'an Jiaotong University, 710061 Xi'an, China.
⁵ Shaanxi Provincial Centre for Disease Control and Prevention, 710054 Xi'an, China.
⁶ Section for Medical Inflammation Research, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm 171 77, Sweden.

Abstract

Coronavirus disease 2019 is a kind of viral pneumonia caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, the mechanism whereby SARS-CoV-2 invades host cells remains poorly understood. Here we used SARS-CoV-2 pseudoviruses to infect human angiotensin-converting enzyme 2 (ACE2) expressing HEK293T cells and evaluated virus infection. We confirmed that SARS-CoV-2 entry was dependent on ACE2 and sensitive to pH of endosome/lysosome in HEK293T cells. The infection of SARS-CoV-2 pseudoviruses is independent of dynamin, clathrin, caveolin and endophilin A2, as well as macropinocytosis. Instead, we found that the infection of SARS-CoV-2 pseudoviruses was cholesterol-rich lipid raft dependent. Cholesterol depletion of cell membranes with methyl-β-cyclodextrin resulted in reduction of pseudovirus infection. The infection of SARS-CoV-2 pseudoviruses resumed with cholesterol supplementation. Together, cholesterol-rich lipid rafts, and endosomal acidification, are key steps of SARS-CoV-2 required for infection of host cells. Therefore, our finding expands the understanding of SARS-CoV-2 entry mechanism and provides a new anti-SARS-CoV-2 strategy.

Keywords: Cholesterol; Endocytosis; Endosomal acidification; Lipid rafts; SARS-CoV-2.