Ex vivo assay to evaluate the efficacy of drugs targeting sphingolipids in preventing SARS-CoV-2 infection of nasal epithelial cells

Katrin Anne Becker; Alexander Carpinteiro; Markus Hoffmann; Stefan Pöhlmann; Johannes Kornhuber; Erich Gulbins

doi:10.1016/j.xpro.2021.100356

Ex vivo assay to evaluate the efficacy of drugs targeting sphingolipids in preventing SARS-CoV-2 infection of nasal epithelial cells

STAR Protoc. 2021 Mar 19;2(1):100356. doi: 10.1016/j.xpro.2021.100356. Epub 2021 Feb 3.

Authors

Katrin Anne Becker¹, Alexander Carpinteiro^{1

2}, Markus Hoffmann^{3

4}, Stefan Pöhlmann^{3

4}, Johannes Kornhuber⁵, Erich Gulbins^{1

6}

Affiliations

¹ Institute of Molecular Biology, University of Duisburg-Essen, Hufelandstrasse 55, 45122 Essen, Germany.
² Department of Hematology, University Hospital Essen, University of Duisburg-Essen, Hufelandstrasse 55, 45122 Essen, Germany.
³ Infection Biology Unit, German Primate Center - Leibniz Institute for Primate Research, Göttingen, Germany.
⁴ Faculty of Biology and Psychology, University of Göttingen, 37073 Göttingen, Germany.
⁵ Department of Psychiatry and Psychotherapy, Friedrich-Alexander University Erlangen-Nürnberg (FAU), Schwabachanlage 6, 91054 Erlangen, Germany.
⁶ Department of Surgery, University of Cincinnati Medical School, 231 Albert Sabin Way, ML0558, Cincinnati, OH 45267, USA.

Abstract

This protocol enables the testing of drugs against infection of epithelial cells with SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2), using pseudo-typed replication deficient vesicular stomatitis virus particles (pp-VSV) presenting the SARS-CoV-2 spike protein. After treating human volunteers with amitriptyline, an approved antidepressant and inhibitor of the acid sphingomyelinase, freshly isolated nasal epithelial cells were infected ex vivo and infection levels were quantified. This protocol offers the possibility to rapidly test the efficacy of potential drugs in the fight against COVID-19. For complete details on the use and execution of this protocol, please refer to Carpinteiro et al. (2020).

Keywords: Cell isolation; Cell-based assays.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antiviral Agents / pharmacology*
COVID-19 / prevention & control*
Cell Culture Techniques
Cells, Cultured
Drug Evaluation, Preclinical / methods*
Epithelial Cells / cytology
Humans
Nasal Mucosa / cytology
SARS-CoV-2 / drug effects*
Sphingolipids / metabolism*
Spike Glycoprotein, Coronavirus
Vesicular stomatitis Indiana virus

Substances

Antiviral Agents
Sphingolipids
Spike Glycoprotein, Coronavirus
spike protein, SARS-CoV-2