Supplementary Materials for Myocyte Specific Upregulation of ACE2 in Cardiovascular Disease: Implications for SARS-CoV-2 mediated myocarditis Nathan R. Tucker,1,2* Mark Chaffin,1* Kenneth C. Bedi Jr.,3 Irinna Papangeli,4 Amer-Denis Akkad,4 Alessandro Arduini,1 Sikander Hayat,4 Gökcen Eraslan,5 Christoph Muus,5,6 Roby Bhattacharyya,5,7 Christian M. Stegmann,4 Human Cell Atlas Lung Biological Network, Kenneth B. Margulies,3 Patrick T. Ellinor1,7 1. Precision Cardiology Laboratory, The Broad Institute, Cambridge, MA, USA 02142 2. Masonic Medical Research Institute, Utica, NY, USA 13501 3. Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA 19104 4. Precision Cardiology Laboratory, Bayer US LLC, Cambridge, MA, USA 02142 5. The Broad Institute of MIT and Harvard, Cambridge, MA, USA 02142 6. John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA, USA 02138 7. Infectious Diseases Division, Department of Medicine, Massachusetts General Hospital, Boston, MA 02114 8. Cardiovascular Research Center, Massachusetts General Hospital, Boston, MA, USA 02114 * These authors contributed equally Running Title: Myocyte upregulation of ACE2 in cardiovascular disease Keywords: COVID-19, SARS-CoV-2, heart, single cell sequencing, myocarditis, cardiovascular disease Corresponding Author: Patrick T. Ellinor, MD, PhD The Broad Institute of MIT and Harvard 75 Ames Street Cambridge, MA 02142 [email protected] Index for Supplemental Materials Item Page HCA Lung Biological Network Consortium Members 3 Supplemental Methods 8 Supplemental Tables 1. Baseline characteristics of study participants 12 2. Differential expression of ACE2 in left ventricular cell subtypes from individuals with dilated cardiomyopathy versus non-failing 13 controls 3. Differential expression of ACE2 in left ventricular cell subtypes from individuals with hypertrophic cardiomyopathy versus non- 14 failing controls 4. Sensitivity analysis comparing the expression of ACE2 in left ventricles samples from individuals with hypertrophic cardiomyopathy compared to non-failing controls after 15 exclusion of individuals on ACE inhibitors or Angiotensin II Receptor Blockers. 5. Comparison of the expression of ACE2 in cell subtypes of left ventricular tissue in the subset of patients with hypertrophic 16 cardiomyopathy that were (ACEi) or were not taking (No ACEi) an ACEi at the time of transplantation Supplemental Figure 1. Cell type resolved expression of SARS-CoV-2 associated proteases TMPRSS1 and CTSL in left ventricular tissue from non- 17 failing controls References 18 HCA Lung Biological Network Consortium Members First name Last name Affiliation Nicholas E. Banovich Translational Genomics Research Institute, Phoenix, AZ. Pascal Barbry Université Côte d’Azur, CNRS, IPMC, Sophia-Antipolis, 06560, France Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital, Boston, Katharine E. Black MA 02114 European Molecular Biology Laboratory, European Bioinformatics Institute (EMBL-EBI), Alvis Brazma Wellcome Trust Genome Campus, Hinxton, Cambridge, CB10 1SD, UK Biosciences Institute, Faculty of Medical Sciences, Newcastle University, International Centre for Joseph Collin Life, Bioscience West Building, Newcastle upon Tyne NE1 3 BZ, UK Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt- Christian Conrad Universität zu Berlin, and Berlin Institute of Health, Charitéplatz 1, 10117 Berlin, Germany Department of Medicine and Institute for Stem Cell Biology and Regenerative Medicine, Tushar Desai Stanford University School of Medicine, Stanford, CA 94116 University of California San Diego and Rady Children’s Hospital San Diego, Department of Thu Elizabeth Duong Pediatrics Division of Respiratory Medicine Division of Pulmonary Sciences and Critical Care Medicine, Department of Medicine, University Oliver Eickelberg of Colorado, Anschutz Medical Campus, Aurora, CO, US Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt- Roland Eils Universität zu Berlin, and Berlin Institute of Health, Charitéplatz 1, 10117 Berlin, Germany Respiratory Bioinformatics and Molecular Biology, University of Technology Sydney, Sydney, Alen Faiz New South Wales, Australia. Institute of Transplant Immunology, Hannover Medical School, MHH, Carl-Neuberg Str. 1, 30625 Hannover, Germany, phone +40 511 532 9745; fax +40 511 532 8090; German Center for Christine Falk Infectious Diseases DZIF, TTU-IICH 07.801 Department of Immunology and Microbiology, The Scripps Research Institute, Jupiter, Florida, Michael Farzan USA (33458) Ian Glass Department of Pediatrics, Genetic Medicine, University of Washington, Seattle, Washington; Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge CB10 1SA, UK; Biosciences Institute, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne NE2 4HH, UK; Department of Dermatology and NIHR Newcastle Biomedical Research Centre, Muzlifa Haniffa Newcastle Hospitals NHS Foundation Trust, Newcastle upon Tyne NE2 4LP, UK. Synthetic and Systems Biology Unit, Biological Research Centre (BRC), Szeged, Hungary; Institute Peter Horvath for Molecular Medicine Finland, University of Helsinki Cardiovascular and Metabolic Sciences, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), 13125 Berlin, Germany; DZHK (German Centre for Cardiovascular Research), Partner Site Berlin, 13347 Berlin, Germany; Berlin Institute of Health (BIH), 10178 Norbert Hubner Berlin, Germany; Charité-Universitätsmedizin, 10117 Berlin, Germany Professor of Genetics, Department of Genetics at Harvard Medical School and Department of Molecular Biology at Massachusetts General Hospital; Co-Director, Infectious Disease and Deborah Hung Microbiome Program and Core Faculty Member, Broad Institute of MIT & Harvard Naftali Kaminski Pulmonary, Critical Care and Sleep Medicine, Yale University School of Medicine Department of Pediatric Pulmonology and Pediatric Allergology, Beatrix Children’s Hospital, University of Groningen, University Medical Center Groningen (UMCG), Groningen Research Gerard Koppelman Institute for Asthma and COPD, Groningen, Netherlands Mark Krasnow Department of Biochemistry and Wall Center for Pulmonary Vascular Disease Division of Allergy, Pulmonary and Critical Care Medicine, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN; Department of Veterans Affairs Medical Center, Nashville, TN; Department of Cell and Developmental Biology, Vanderbilt University, Nashville, Jonathan A. Kropski TN Malte Kuhnemund Cartana AB, Nobels vag 16, 17165 Stockholm, Sweden Division of Rheumatology, Department of Medicine, University of Pittsburgh Medical Center, Robert Lafyatis Pittsburgh, PA, USA. Biosciences Institute, Faculty of Medical Sciences, Newcastle University, International Centre for Majlinda Lako Life, Bioscience West Building, Newcastle upon Tyne NE1 3 BZ, UK Haeock Lee Department of Biomedicine and Health Sciences, The Catholic University of Korea, Seoul, Korea Université Côte d'Azur, CHU de Nice, FHU OncoAge, Department of Pulmonary Medicine and Allergology, Nice, France ; CNRS UMR 7275 - Institut de Pharmacologie Moléculaire et Cellulaire, Sylvie Leroy Sophia Antipolis, France Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Sten Linnarson Karolinska Institute Joakim Lundeberg SciLifeLab, Department of Gene Technology, KTH Royal Institute of Technology Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital, Boston, Benjamin Medoff MA 02114 Kerstin Meyer Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK Chichau Miao Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge CB10 1SA, UK Alexander Misharin Division of Pulmonary and Critical Care Medicine, Northwestern University, Chicago, Illinois Department of Pathology and Medical Biology, University of Groningen, GRIAC Research Martijn Nawijn institue, University Medical Center Groningen, the Netherlands Marko Nikolic UCL Respiratory, Division of Medicine, University College London, London, UK National Heart and Lung Institute, Imperial College London, UK; British Heart Foundation Centre Michela Noseda for Research Excellence and Centre for Regenerative Medicine, Imperial College London, UK Division of Gastroenterology Boston Children's Hospital, Boston, MA, USA; Program in Ordovas Immunology, Harvard Medical School, Boston, MA, USA; Broad Institute of MIT and Harvard, Jose Montanes Cambridge, MA, USA; Harvard Stem Cell Institute, Cambridge, MA, USA. Canada Research Chair in Heart Failure, Division of Cardiology, 2C2 Walter Mackenzie Health Gavin Oudit Sciences Centre, Edmonton, Alberta, T6G 2B7, Canada Computational and Systems Biology Program, Sloan Kettering Institute, Memorial Sloan Dana Pe'er Kettering Cancer Center, New York, New York, USA Garvan-Weizmann Centre for Cellular Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia; UNSW Cellular Genomics Futures Institute, University of New South Wales, Joseph Powell Sydney, NSW, Australia Depts of Bioengineering and Applied Physics, Stanford University, and the Chan Zuckerberg Steve Quake Biohub. Harvard Stem Cell Institute, Cambridge, Massachusetts; Center for Regenerative Medicine, Jay Rajagopal Massachusetts General Hospital, Boston, Massachusetts Department of Cell Biology, Regeneration Next Initiative, Duke University School of Medicine, Purushothama