Patients with cardiovascular comorbidities are more susceptible to severe infection with SARS-CoV-2, known to directly cause pathological damage to cardiovascular tissue. We outline a screening platform using human embryonic stem cell-derived cardiomyocytes, confirmed to express the protein machinery critical for SARS-CoV-2 infection, and a SARS-CoV-2 spike-pseudotyped virus system. The method has allowed us to identify benztropine and DX600 as novel inhibitors of SARS-CoV-2 infection in a clinically relevant stem cell-derived cardiomyocyte line. Discovery of new medicines will be critical for protecting the heart in patients with SARS-CoV-2, and for individuals where vaccination is contraindicated

Bloor, Stuart

Colzani, Maria T

Davenport, Anthony P

Greenwood, Edward J D

Kuc, Rhoda E

Lehner, Paul J

Macrae, Robyn G C

Maguire, Janet J

Nyimanu, Duuamene

Robinson, Emma L

Sinha, Sanjay

Strachan, Gregory

Williams, Thomas L

Zhan, Jun Ru

Communications Biology

English

Williams et al. confirm that human embryonic stem cell-derived cardiomyocytes express the protein machinery critical for SARS-CoV-2 infection and are susceptible to SARS-CoV-2 spike protein pseudotyped virus infection. They further use this platform as a screen to identify inhibitors of SARS-CoV-2 infection, reporting benztropine (targeting B0AT1/ACE2 complex) and DX600 (targeting ACE2) as potential inhibitors

Thomas L. Williams

Maria T. Colzani

Robyn G. C. Macrae

Emma L. Robinson

Stuart Bloor

Edward J. D. Greenwood

Jun Ru Zhan

Gregory Strachan

Rhoda E. Kuc

Duuamene Nyimanu

Janet J. Maguire

Paul J. Lehner

Sanjay Sinha

Anthony P. Davenport

Directory of Open Access Journals

Human embryonic stem cell-derived cardiomyocyte platform screens inhibitors of SARS-CoV-2 infection

Apollo (Cambridge)

Human embryonic stem cell-derived cardiomyocyte platform screens inhibitors of SARS-CoV-2 infection.

Patients with cardiovascular comorbidities are more susceptible to severe infection with SARS-CoV-2, known to directly cause pathological damage to cardiovascular tissue. We outline a screening platform using human embryonic stem cell-derived cardiomyocytes, confirmed to express the protein machinery critical for SARS-CoV-2 infection, and a SARS-CoV-2 spike-pseudotyped virus system. The method has allowed us to identify benztropine and DX600 as novel inhibitors of SARS-CoV-2 infection in a clinically relevant stem cell-derived cardiomyocyte line. Discovery of new medicines will be critical for protecting the heart in patients with SARS-CoV-2, and for individuals where vaccination is contraindicated.This research was funded in whole, or in part by: Wellcome Trust (WT107715/Z/15/Z, A.P.D., J.J.M.);  Addenbrooke’s Charitable Trust (P.J.L., A.P.D.);  Wellcome Trust Programme in Metabolic and Cardiovascular Disease (203814/Z/16/A, T.L.W., D.N.),  Wellcome Trust Major Award (208363/Z/17/Z) for Imaging Core (G.S.); Wellcome Trust Principal Research Fellowship (210688/Z/18/Z, P.J.L.), UKRI/NIHR through the UK Coronavirus Immunology Consortium (P.J.L.); Rosetrees Trust Charity (G103718, J.R.Z.);  British Heart Foundation (FS/17/61/33473 APD, S.S., R.G.C.M; TAF 03, APD, JJM);  British Heart Foundation Senior Fellowship (FS/18/46/33663, S.S.), British Heart Foundation Centre for Regenerative Medicine (RM/17/2/33380, S.S., M.C.); Cambridge Biomedical Research Centre Biomedical Resources Grant (University of Cambridge, Cardiovascular Theme, RG64226) and a core support grant from the Wellcome Trust and MRC to the Wellcome–MRC Cambridge Stem Cell Institute

Macrae, Robyn GC

Greenwood, Edward JD

Funder: Cambridge Biomedical Research Centre Biomedical Resources Grant (University of Cambridge, Cardiovascular Theme RG64226Abstract: Patients with cardiovascular comorbidities are more susceptible to severe infection with SARS-CoV-2, known to directly cause pathological damage to cardiovascular tissue. We outline a screening platform using human embryonic stem cell-derived cardiomyocytes, confirmed to express the protein machinery critical for SARS-CoV-2 infection, and a SARS-CoV-2 spike-pseudotyped virus system. The method has allowed us to identify benztropine and DX600 as novel inhibitors of SARS-CoV-2 infection in a clinically relevant stem cell-derived cardiomyocyte line. Discovery of new medicines will be critical for protecting the heart in patients with SARS-CoV-2, and for individuals where vaccination is contraindicated

Williams, Thomas L.

Colzani, Maria T.

Macrae, Robyn G. C.

Robinson, Emma L.

Greenwood, Edward J. D.

Kuc, Rhoda E.

Maguire, Janet J.

Lehner, Paul J.

Davenport, Anthony P.

1  Supplementary Information  Human embryonic stem cell-derived cardiomyocyte platform screens inhibitors of SARS-CoV-2 infection   Thomas L. Williams1§, Maria T. Colzani2§, Robyn G.C. Macrae1,2, Emma L. Robinson3, Stuart Bloor4, Edward J. D. Greenwood4, Jun Ru Zhan4,  Gregory Strachan5, Rhoda E. Kuc1, Duuamene Nyimanu1,  Janet J. Maguire1, Paul J. Lehner4, Sanjay Sinha2#, Anthony P. Davenport1#*  1. Experimental Medicine and Immunotherapeutics, University of Cambridge, Addenbrooke’s Hospital, Cambridge, U.K. 2. Wellcome-MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, U.K.  3. School of Medicine, Division of Cardiology, University of Colorado Denver, Anschutz Medical Campus, Aurora, U.S.A. 4. Cambridge Institute of Therapeutic Immunology & Infectious Disease (CITIID), Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, U.K. 5. Wellcome Trust-MRC Institute of Metabolic Science, Metabolic Research Laboratories, Addenbrooke’s Biomedical Campus, Cambridge, U.K.  §Joint first authors, contributed equally. #Joint senior authors.   Author for Correspondence: *Anthony P. Davenport, Experimental Medicine and Immunotherapy,                         University of Cambridge, Level 6, Addenbrooke’s Centre for Clinical Investigation,                                                Box 110, Addenbrooke's Hospital,             Cambridge, CB2 0QQ, U.K.                Tel:+44(0)1223 336899                                                       apd10@medschl.cam.ac.uk                            2  Supplementary Fig. 1: Titre- and time-dependent SARS-CoV-2 infection of hESC-derived cardiomyocytes. Representative fluorescent images (n=2 independent experiments) of human embryonic stem cell-derived cardiomyocytes (hESC-CMs) infected with SARS-CoV-2 at several different viral titres ((0.001-0.1 multiplicity of infection (MOI)), or left untreated, and incubated for different time periods (24-72 h). Infected cells were immunolabelled after fixation with 2% formaldehyde and permeabilization with BD Perm/Wash buffer (BD Biosciences, 554723) using sheep anti-SARS-CoV-2 nucleocapsid antibody (DA114, MRC-PPU). Cells were visualised using a donkey anti-sheep secondary antibody conjugated to Alexa Fluor 488 (green) (Jackson ImmunoResearch #713-545-147).                               3      Supplementary Fig. 2: Cell populations of pure, beating cardiomyocytes. a-e Representative brightfield video files of beating human embryonic stem cell-derived cardiomyocytes (hESC-CMs) following infection with 25 μL (a), 50 μL (b), or 100 μL (c) SARS-CoV-2 spike pseudotyped lentivirus. Control cells were treated with 100 μL vesicular stomatitis virus (VSV-G) pseudotyped lentivirus (d) or left untreated with viral particles (e). f-i hESC purity flow cytometry data. f Cells were gated using FSC and SSC. g Single cells were distinguished from doublets and aggregates by plotting FSC-A against FSC-W. Subsequent histograms were gated on the single cells population. h The Troponin-T gate was determined using cells stained with IgG control. i Representative plot displaying purity of hESC-CM differentiation by flow cytometry using an antibody specific for cardiac troponin-T (96.8 % of the population shown to be positive for this marker).    4    Supplementary Table. 1: Compounds used, their protein targets, and other relevant information. The table shows the compounds used in screening experiments at the denoted concentrations, and their respective protein targets shown to be expressed in the human embryonic stem cell-derived cardiomyocytes (hESC-CMs) cell model, as well as adult cardiomyocytes. Concentrations used in the hESC-CM screen were chosen based on the pIC50 values reported in literature or on IUPHAR/BPS Guide to PHARMACOLOGY (https://www.guidetopharmacology.org). 

https://www.repository.cam.ac.uk/bitstream/handle/1810/326047/42003_2021_2453_MOESM1_ESM.pdf?sequence=3&isAllowed=y

Human embryonic stem cell-derived cardiomyocyte platform screens inhibitors of SARS-CoV-2 infection.

Abstract

Similar works

Full text

Available Versions

Directory of Open Access Journals

Apollo (Cambridge)

Apollo (Cambridge)

Apollo (Cambridge)