Parallel use of human stem cell lung and heart models provide insights for SARS-CoV-2 treatment
Rajeev Rudraraju, Matthew J Gartner, Jessica A Neil, Elizabeth S Stout, Joseph Chen, Elise J Needham, Michael See, Charley Mackenzie-Kludas, Leo Yi Yang Lee, Mingyang Wang, Hayley Pointer, Kathy Karavendzas, Dad Abu-Bonsrah, Damien Drew, Yu Bo Yang Sun, Jia Ping Tan, Guizhi Sun, Abbas Salavaty, Natalie Charitakis, Hieu T Nim, Peter D Currie, Wai-Hong Tham, Enzo Porrello, Jose M Polo, Sean J Humphrey, Mirana Ramialison, David A Elliott, Kanta Subbarao
Stem Cell Reports, doi:10.1016/j.stemcr.2023.05.007
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) primarily infects the respiratory tract, but pulmonary and cardiac complications occur in severe coronavirus disease 2019 . To elucidate molecular mechanisms in the lung and heart, we conducted paired experiments in human stem cell-derived lung alveolar type II (AT2) epithelial cell and cardiac cultures infected with SARS-CoV-2. With CRISPR-Cas9-mediated knockout of ACE2, we demonstrated that angiotensin-converting enzyme 2 (ACE2) was essential for SARS-CoV-2 infection of both cell types but that further processing in lung cells required TMPRSS2, while cardiac cells required the endosomal pathway. Host responses were significantly different; transcriptome profiling and phosphoproteomics responses depended strongly on the cell type. We identified several antiviral compounds with distinct antiviral and toxicity profiles in lung AT2 and cardiac cells, highlighting the importance of using several relevant cell types for evaluation of antiviral drugs. Our data provide new insights into rational drug combinations for effective treatment of a virus that affects multiple organ systems.
EXPERIMENTAL PROCEDURES
Resource availability Corresponding authors Enzo R. Porrello, Jose M. Polo, Sean J. Humphrey, Mirana Ramialison, David A. Elliott or Kanta Subbarao are the corresponding authors for this paper.
Cells African green monkey kidney epithelial (Vero cells, ATCC Cat. CCL-81), Vero hSLAM (Merck, Cat. 04091501), Calu-3 (ATCC, Cat. HTB-55), and VeroE6-TMPRSS2 (CellBank Australia, Cat. JCRB1819) cells were cultured at 37 C and 5% CO 2. Vero cell media: minimum essential medium (MEM; Media Preparation Unit, Peter Doherty Institute) supplemented with 5% fetal bovine serum (FBS; Bovogen, Cat. SFBS), 50 U/mL penicillin and 50 mg/mL streptomycin (PenStrep, Thermo Fisher Scientific, Cat. 15070-063), 2 mM GlutaMAX (Thermo Fisher Scientific, Cat. 35050061), and 15 mM HEPES (Thermo Fisher Scientific, Cat. 15630130) . Vero hSLAM cell media: MEM supplemented with 7% FBS, PenStrep, 2 mM GlutaMAX, 15 mM HEPES, and 0.4 mg/mL G418 Sulfate (Gibco, Cat. 10131027). Calu-3 cell media: MEM containing L-glutamine and sodium bicarbonate (Sigma, Cat. M4655) supplemented with 10% FBS, PenStrep, 13 non-essential amino acids (Gibco, Cat. 11140050), and sodium pyruvate (Fisher Scientific, Cat. BP356-100). VeroE6-TMPRSS2 cell media: Dulbecco's MEM (DMEM; Media Preparation Unit, Peter Doherty Institute) supplemented with 10% FBS, PenStrep, 2 mM GlutaMAX, and 1 mg/ mL G418 Sulfate. (Hou et al., 2020) . SARS-CoV-2 VIC01 was propagated in Vero and Vero hSLAM cells in Vero infection..
References
Anderson, Kaplan, Bell, Koutsis, Haynes et al., NKX2-5 regulates human cardiomyogenesis via a HEY2 dependent transcriptional network, Nat. Commun,
doi:10.1038/s41467-018-03714-xBailey, Dmytrenko, Greenberg, Bredemeyer, Ma et al., SARS-CoV-2 infects human engineered heart tissues and models COVID-19 myocarditis, JACC Basic Transl Sci,
doi:10.1016/j.jacbts.2021.01.002Bojkova, Wagner, Shumliakivska, Aslan, Saleem et al., SARS-CoV-2 infects and induces cytotoxic effects in human cardiomyocytes, Cardiovasc. Res,
doi:10.1093/cvr/cvaa267Chen, Bobrovitz, Premji, Koopmans, Fisman et al., SARS-CoV-2 shedding dynamics across the respiratory tract, sex, and disease severity for adult and pediatric COVID-19, Elife,
doi:10.7554/eLife.70458Chen, Gao, Wang, Wei, An iTSCderived placental model of SARS-CoV-2 infection reveals ACE2-dependent susceptibility in syncytiotrophoblasts, bioRxiv,
doi:10.1101/2021.10.27.465224Deinhardt-Emmer, Wittschieber, Sanft, Kleemann, Elschner et al., Early postmortem mapping of SARS-CoV-2 RNA in patients with COVID-19 and the correlation with tissue damage, Elife,
doi:10.7554/eLife.60361Elliott, Braam, Koutsis, Ng, Jenny et al., NKX2-5(eGFP/w) hESCs for isolation of human cardiac progenitors and cardiomyocytes, Nat. Methods,
doi:10.1038/nmeth.1740Goyal, Choi, Pinheiro, Schenck, Chen et al., Clinical characteristics of Covid-19 in New York City, N. Engl. J. Med,
doi:10.1056/NEJMc2010419Gunst, Staerke, Pahus, Kristensen, Bodilsen et al., Efficacy of the TMPRSS2 inhibitor camostat mesilate in patients hospitalized with Covid-19-a double-blind randomized controlled trial, EClinicalMedicine,
doi:10.1016/j.eclinm.2021.100849Hamming, Timens, Bulthuis, Lely, Navis et al., Tissue distribution of ACE2 protein, the functional receptor for SARS coronavirus. A first step in understanding SARS pathogenesis, J. Pathol,
doi:10.1002/path.1570Hoffmann, Kleine-Weber, Schroeder, Kru ¨ger, Herrler et al., SARS-CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor, Cell,
doi:10.1016/j.cell.2020.05.042Huang, Hume, Abo, Werder, Villacorta-Martin et al., SARS-CoV-2 infection of pluripotent stem cell-derived human lung alveolar type 2 cells elicits a rapid epithelial-intrinsic inflammatory response, Cell Stem Cell,
doi:10.1016/j.stem.2020.09.013Humphrey, Karayel, James, Mann, High-throughput and high-sensitivity phosphoproteomics with the EasyPhos platform, Nat. Protoc,
doi:10.1038/s41596-018-0014-9Jacob, Morley, Hawkins, Mccauley, Jean et al., Differentiation of human pluripotent stem cells into functional lung alveolar epithelial cells, Cell Stem Cell,
doi:10.1016/j.stem.2017.08.014Katsura, Sontake, Tata, Kobayashi, Edwards et al., Human lung stem cell-based alveolospheres provide insights into SARS-CoV-2-mediated interferon responses and pneumocyte dysfunction, Cell Stem Cell,
doi:10.1016/j.stem.2020.10.005Lazzerini, Laghi-Pasini, Boutjdir, Capecchi, Inflammatory cytokines and cardiac arrhythmias: the lesson from COVID-19, Nat. Rev. Immunol,
doi:10.1038/s41577-022-00714-3Lee, Huang, Rastegari, Rengganaten, Liu et al., Tumor necrosis factor-alpha exacerbates viral entry in SARS-CoV2-infected iPSC-derived cardiomyocytes, Int. J. Mol. Sci,
doi:10.3390/ijms22189869Li, Renner, Comar, Whelan, Reyes et al., SARS-CoV-2 induces double-stranded RNAmediated innate immune responses in respiratory epithelialderived cells and cardiomyocytes,
doi:10.1073/pnas.2022643118Liao, Smyth, Shi, featureCounts: an efficient general purpose program for assigning sequence reads to genomic features, Bioinformatics,
doi:10.1093/bioinformatics/btt656Lopes, Garcia-Herna ´ndez, Lorenzini, Futema, Chumakova et al., Alpha-protein kinase 3 (ALPK3) truncating variants are a cause of autosomal dominant hypertrophic cardiomyopathy, Eur. Heart J,
doi:10.1093/eurheartj/ehab424Marchiano, Hsiang, Khanna, Higashi, Whitmore et al., SARS-CoV-2 infects human pluripotent stem cellderived cardiomyocytes, impairing electrical and mechanical function, Stem Cell Rep,
doi:10.1016/j.stemcr.2021.02.008Mills, Titmarsh, Koenig, Parker, Ryall et al., Functional screening in human cardiac organoids reveals a metabolic mechanism for cardiomyocyte cell cycle arrest, Proc. Natl. Acad. Sci. USA,
doi:10.1073/pnas.1707316114Montaser, Lalmanach, Mach, CA-074, but not its methyl ester CA-074Me, is a selective inhibitor of cathepsin B within living cells, Biol. Chem,
doi:10.1515/BC.2002.147Mulay, Konda, Garcia, Jr, Yao et al., SARS-CoV-2 infection of primary human lung epithelium for COVID-19 modeling and drug discovery, Cell Rep,
doi:10.1016/j.celrep.2021.109055Muus, Luecken, Eraslan, Sikkema, Waghray et al., Single-cell meta-analysis of SARS-CoV-2 entry genes across tissues and demographics, Nat. Med,
doi:10.1038/s41591-020-01227-zNavaratnarajah, Pease, Halfmann, Taye, Barkhymer et al., Highly efficient SARS-CoV-2 infection of human cardiomyocytes: spike protein-mediated cell fusion and its inhibition, J. Virol,
doi:10.1128/JVI.01368-21Ou, Liu, Lei, Li, Mi et al., Characterization of spike glycoprotein of SARS-CoV-2 on virus entry and its immune cross-reactivity with SARS-CoV, Nat. Commun,
doi:10.1038/s41467-020-15562-9Perez-Bermejo, Kang, Rockwood, Simoneau, Joy et al., SARS-CoV-2 infection of human iPSC-derived cardiac cells reflects cytopathic features in hearts of patients with COVID-19, Sci. Transl. Med,
doi:10.1126/scitranslmed.abf7872Phelan, Anderson, Howden, Wong, Hickey et al., ALPK3-deficient cardiomyocytes generated from patientderived induced pluripotent stem cells and mutant human embryonic stem cells display abnormal calcium handling and establish that ALPK3 deficiency underlies familial cardiomyopathy, Eur. Heart J,
doi:10.1093/eurheartj/ehw160Pruijssers, George, Scha ¨fer, Leist, Gralinksi et al., Remdesivir inhibits SARS-CoV-2 in human lung cells and chimeric SARS-CoV expressing the SARS-CoV-2 RNA polymerase in mice, Cell Rep,
doi:10.1016/j.celrep.2020.107940Puntmann, Carerj, Wieters, Fahim, Arendt et al., Outcomes of cardiovascular magnetic resonance imaging in patients recently recovered from coronavirus disease 2019 (COVID-19), JAMA Cardiol,
doi:10.1001/jamacardio.2020.3557Qi, Qian, Zhang, Zhang, Single cell RNA sequencing of 13 human tissues identify cell types and receptors of human coronaviruses, Biochem. Biophys. Res. Commun,
doi:10.1016/j.bbrc.2020.03.044Robinson, Alkass, Bergmann, Maguire, Roderick et al., Genes encoding ACE2, TMPRSS2 and related proteins mediating SARS-CoV-2 viral entry are upregulated with age in human cardiomyocytes, J. Mol. Cell. Cardiol,
doi:10.1016/j.yjmcc.2020.08.009Robinson, Mccarthy, Smyth, edgeR: a Bioconductor package for differential expression analysis of digital gene expression data, Bioinformatics,
doi:10.1093/bioinformatics/btp616Schneider, Pease, Navaratnarajah, Halfmann, Clemens et al., SARS-CoV-2 direct cardiac damage through spike-mediated cardiomyocyte fusion
Shang, Wan, Luo, Ye, Geng et al., Cell entry mechanisms of SARS-CoV-2, Proc. Natl. Acad. Stem Cell Reports j
Shao, Shang, Luo, Shi, Zhao et al., Myocardial injury is associated with higher mortality in patients
Sharma, Garcia, Jr, Wang, Plummer et al., Human iPSCderived cardiomyocytes are susceptible to SARS-CoV-2 infection, Cell Rep. Med,
doi:10.1016/j.xcrm.2020.100052Shemesh, Aktepe, Deerain, Mcauley, Audsley et al., SARS-CoV-2 suppresses IFNbeta production mediated by NSP1, 5, 6, 15, ORF6 and ORF7b but does not suppress the effects of added interferon, PLoS Pathog,
doi:10.1371/journal.ppat.1009800Shi, Qin, Shen, Cai, Liu et al., Association of cardiac injury with mortality in hospitalized patients with COVID-19 in Wuhan, JAMA Cardiol,
doi:10.1001/jamacardio.2020.0950Sungnak, Huang, Be ´cavin, Berg, Queen et al., SARS-CoV-2 entry factors are highly expressed in nasal epithelial cells together with innate immune genes, Nat. Med,
doi:10.1038/s41591-020-0868-6Tiwari, Wang, Smith, Carlin, Revealing tissue-specific SARS-CoV-2 infection and host responses using human stem cell-derived lung and cerebral organoids, Stem Cell Rep,
doi:10.1016/j.stemcr.2021.02.005Tohyama, Hattori, Sano, Hishiki, Nagahata et al., Distinct metabolic flow enables large-scale purification of mouse and human pluripotent stem cell-derived cardiomyocytes, Cell Stem Cell,
doi:10.1016/j.stem.2012.09.013Uhle ´n, Fagerberg, Hallstro ¨m, Lindskog, Oksvold et al., Proteomics. Tissue-based map of the human proteome, Science,
doi:10.1126/science.1260419Wang, Mannan, Xiao, Abdulfatah, Qiao et al., Characterization of SARS-CoV-2 and host entry factors distribution in a COVID-19 autopsy series, Commun. Med. 1,
doi:10.1038/s43856-021-00025-zWilliams, Colzani, Macrae, Robinson, Bloor et al., Human embryonic stem cell-derived cardiomyocyte platform screens inhibitors of SARS-CoV-2 infection, Commun. Biol,
doi:10.1038/s42003-021-02453-yYaron, Heaton, Levy, Johnson, Jordan et al., The FDA-approved drug Alectinib compromises SARS-CoV-2 nucleocapsid phosphorylation and inhibits viral infection in vitro, bioRxiv,
doi:10.1101/2020.08.14.251207Youk, Kim, Evans, Jeong, Hur et al., Three-dimensional human alveolar stem cell culture models reveal infection response to SARS-CoV-2, Cell Stem Cell,
doi:10.1016/j.stem.2020.10.004Zhang, Bastard, Liu, Le Pen, Moncada-Velez et al., Inborn errors of type I IFN immunity in patients with life-threatening COVID-19, Science,
doi:10.1126/science.abd4570Zhou, Yang, Wang, Hu, Zhang et al., Addendum: a pneumonia outbreak associated with a new coronavirus of probable bat origin, Nature,
doi:10.1038/s41586-020-2951-zZhou, Zhou, Pache, Chang, Khodabakhshi et al., Metascape provides a biologist-oriented resource for the analysis of systems-level datasets, Nat. Commun,
doi:10.1038/s41467-019-09234-6Zou, Chen, Zou, Han, Hao et al., Single-cell RNA-seq data analysis on the receptor ACE2 expression reveals the potential risk of different human organs vulnerable to 2019-nCoV infection, Front. Med,
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'doi-asserted-by': 'crossref',
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'volume': '6',
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'volume': '116',
'author': 'Bojkova',
'year': '2020',
'journal-title': 'Cardiovasc. Res.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib4',
'doi-asserted-by': 'crossref',
'first-page': '685',
'DOI': '10.1016/j.cell.2020.06.034',
'article-title': 'The global phosphorylation landscape of SARS-CoV-2 infection',
'volume': '182',
'author': 'Bouhaddou',
'year': '2020',
'journal-title': 'Cell'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib5',
'article-title': 'An iTSC-derived placental model of SARS-CoV-2 infection reveals '
'ACE2-dependent susceptibility in syncytiotrophoblasts',
'author': 'Chen',
'year': '2021',
'journal-title': 'bioRxiv'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib6',
'doi-asserted-by': 'crossref',
'first-page': 'e70458',
'DOI': '10.7554/eLife.70458',
'article-title': 'SARS-CoV-2 shedding dynamics across the respiratory tract, sex, and '
'disease severity for adult and pediatric COVID-19',
'volume': '10',
'author': 'Chen',
'year': '2021',
'journal-title': 'Elife'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib7',
'doi-asserted-by': 'crossref',
'first-page': 'e60361',
'DOI': '10.7554/eLife.60361',
'article-title': 'Early postmortem mapping of SARS-CoV-2 RNA in patients with COVID-19 '
'and the correlation with tissue damage',
'volume': '10',
'author': 'Deinhardt-Emmer',
'year': '2021',
'journal-title': 'Elife'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib8',
'doi-asserted-by': 'crossref',
'first-page': '15',
'DOI': '10.1093/bioinformatics/bts635',
'article-title': 'STAR: ultrafast universal RNA-seq aligner',
'volume': '29',
'author': 'Dobin',
'year': '2013',
'journal-title': 'Bioinformatics'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib9',
'doi-asserted-by': 'crossref',
'first-page': '1037',
'DOI': '10.1038/nmeth.1740',
'article-title': 'NKX2-5(eGFP/w) hESCs for isolation of human cardiac progenitors and '
'cardiomyocytes',
'volume': '8',
'author': 'Elliott',
'year': '2011',
'journal-title': 'Nat. Methods'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib10',
'doi-asserted-by': 'crossref',
'first-page': '2372',
'DOI': '10.1056/NEJMc2010419',
'article-title': 'Clinical characteristics of Covid-19 in New York City',
'volume': '382',
'author': 'Goyal',
'year': '2020',
'journal-title': 'N.\xa0Engl. J. Med.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib11',
'doi-asserted-by': 'crossref',
'first-page': '100849',
'DOI': '10.1016/j.eclinm.2021.100849',
'article-title': 'Efficacy of the TMPRSS2 inhibitor camostat mesilate in patients '
'hospitalized with Covid-19-a double-blind randomized controlled trial',
'volume': '35',
'author': 'Gunst',
'year': '2021',
'journal-title': 'EClinicalMedicine'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib12',
'doi-asserted-by': 'crossref',
'first-page': '631',
'DOI': '10.1002/path.1570',
'article-title': 'Tissue distribution of ACE2 protein, the functional receptor for SARS '
'coronavirus. A first step in understanding SARS pathogenesis',
'volume': '203',
'author': 'Hamming',
'year': '2004',
'journal-title': 'J.\xa0Pathol.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib13',
'doi-asserted-by': 'crossref',
'first-page': '271',
'DOI': '10.1016/j.cell.2020.02.052',
'article-title': 'SARS-CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a '
'clinically proven protease inhibitor',
'volume': '181',
'author': 'Hoffmann',
'year': '2020',
'journal-title': 'Cell'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib14',
'doi-asserted-by': 'crossref',
'first-page': '429',
'DOI': '10.1016/j.cell.2020.05.042',
'article-title': 'SARS-CoV-2 reverse genetics reveals a variable infection gradient in '
'the respiratory tract',
'volume': '182',
'author': 'Hou',
'year': '2020',
'journal-title': 'Cell'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib15',
'doi-asserted-by': 'crossref',
'first-page': 'D884',
'DOI': '10.1093/nar/gkaa942',
'article-title': 'Ensembl 2021',
'volume': '49',
'author': 'Howe',
'year': '2021',
'journal-title': 'Nucleic Acids Res.'},
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'doi-asserted-by': 'crossref',
'first-page': '962',
'DOI': '10.1016/j.stem.2020.09.013',
'article-title': 'SARS-CoV-2 infection of pluripotent stem cell-derived human lung '
'alveolar type 2 cells elicits a rapid epithelial-intrinsic inflammatory '
'response',
'volume': '27',
'author': 'Huang',
'year': '2020',
'journal-title': 'Cell Stem Cell'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib17',
'doi-asserted-by': 'crossref',
'first-page': '1897',
'DOI': '10.1038/s41596-018-0014-9',
'article-title': 'High-throughput and high-sensitivity phosphoproteomics with the '
'EasyPhos platform',
'volume': '13',
'author': 'Humphrey',
'year': '2018',
'journal-title': 'Nat. Protoc.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib18',
'doi-asserted-by': 'crossref',
'first-page': '472',
'DOI': '10.1016/j.stem.2017.08.014',
'article-title': 'Differentiation of human pluripotent stem cells into functional lung '
'alveolar epithelial cells',
'volume': '21',
'author': 'Jacob',
'year': '2017',
'journal-title': 'Cell Stem Cell'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib19',
'doi-asserted-by': 'crossref',
'first-page': '890',
'DOI': '10.1016/j.stem.2020.10.005',
'article-title': 'Human lung stem cell-based alveolospheres provide insights into '
'SARS-CoV-2-mediated interferon responses and pneumocyte dysfunction',
'volume': '27',
'author': 'Katsura',
'year': '2020',
'journal-title': 'Cell Stem Cell'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib20',
'doi-asserted-by': 'crossref',
'first-page': '270',
'DOI': '10.1038/s41577-022-00714-3',
'article-title': 'Inflammatory cytokines and cardiac arrhythmias: the lesson from '
'COVID-19',
'volume': '22',
'author': 'Lazzerini',
'year': '2022',
'journal-title': 'Nat. Rev. Immunol.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib21',
'doi-asserted-by': 'crossref',
'first-page': '9869',
'DOI': '10.3390/ijms22189869',
'article-title': 'Tumor necrosis factor-alpha exacerbates viral entry in '
'SARS-CoV2-infected iPSC-derived cardiomyocytes',
'volume': '22',
'author': 'Lee',
'year': '2021',
'journal-title': 'Int. J. Mol. Sci.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib22',
'doi-asserted-by': 'crossref',
'first-page': '3810',
'DOI': '10.1038/s41467-020-17665-9',
'article-title': 'Activation and evasion of type I interferon responses by SARS-CoV-2',
'volume': '11',
'author': 'Lei',
'year': '2020',
'journal-title': 'Nat. Commun.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib23',
'article-title': 'SARS-CoV-2 induces double-stranded RNA-mediated innate immune responses '
'in respiratory epithelial-derived cells and cardiomyocytes',
'volume': '118',
'author': 'Li',
'year': '2021',
'journal-title': 'Proc. Natl. Acad. Sci. USA'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib24',
'doi-asserted-by': 'crossref',
'first-page': '923',
'DOI': '10.1093/bioinformatics/btt656',
'article-title': 'featureCounts: an efficient general purpose program for assigning '
'sequence reads to genomic features',
'volume': '30',
'author': 'Liao',
'year': '2014',
'journal-title': 'Bioinformatics'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib25',
'doi-asserted-by': 'crossref',
'first-page': '3063',
'DOI': '10.1093/eurheartj/ehab424',
'article-title': 'Alpha-protein kinase 3 (ALPK3) truncating variants are a cause of '
'autosomal dominant hypertrophic cardiomyopathy',
'volume': '42',
'author': 'Lopes',
'year': '2021',
'journal-title': 'Eur. Heart J.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib26',
'doi-asserted-by': 'crossref',
'first-page': '478',
'DOI': '10.1016/j.stemcr.2021.02.008',
'article-title': 'SARS-CoV-2 infects human pluripotent stem cell-derived cardiomyocytes, '
'impairing electrical and mechanical function',
'volume': '16',
'author': 'Marchiano',
'year': '2021',
'journal-title': 'Stem Cell Rep.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib27',
'doi-asserted-by': 'crossref',
'first-page': 'E8372',
'DOI': '10.1073/pnas.1707316114',
'article-title': 'Functional screening in human cardiac organoids reveals a metabolic '
'mechanism for cardiomyocyte cell cycle arrest',
'volume': '114',
'author': 'Mills',
'year': '2017',
'journal-title': 'Proc. Natl. Acad. Sci. USA'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib28',
'doi-asserted-by': 'crossref',
'first-page': '1305',
'DOI': '10.1515/BC.2002.147',
'article-title': 'CA-074, but not its methyl ester CA-074Me, is a selective inhibitor of '
'cathepsin B within living cells',
'volume': '383',
'author': 'Montaser',
'year': '2002',
'journal-title': 'Biol. Chem.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib29',
'doi-asserted-by': 'crossref',
'first-page': '109055',
'DOI': '10.1016/j.celrep.2021.109055',
'article-title': 'SARS-CoV-2 infection of primary human lung epithelium for COVID-19 '
'modeling and drug discovery',
'volume': '35',
'author': 'Mulay',
'year': '2021',
'journal-title': 'Cell Rep.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib30',
'doi-asserted-by': 'crossref',
'first-page': '546',
'DOI': '10.1038/s41591-020-01227-z',
'article-title': 'Single-cell meta-analysis of SARS-CoV-2 entry genes across tissues and '
'demographics',
'volume': '27',
'author': 'Muus',
'year': '2021',
'journal-title': 'Nat. Med.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib31',
'doi-asserted-by': 'crossref',
'first-page': '601',
'DOI': '10.1038/s41591-021-01283-z',
'article-title': 'Post-acute COVID-19 syndrome',
'volume': '27',
'author': 'Nalbandian',
'year': '2021',
'journal-title': 'Nat. Med.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib32',
'doi-asserted-by': 'crossref',
'first-page': 'e0136821',
'DOI': '10.1128/JVI.01368-21',
'article-title': 'Highly efficient SARS-CoV-2 infection of human cardiomyocytes: spike '
'protein-mediated cell fusion and its inhibition',
'volume': '95',
'author': 'Navaratnarajah',
'year': '2021',
'journal-title': 'J.\xa0Virol.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib33',
'doi-asserted-by': 'crossref',
'first-page': '1620',
'DOI': '10.1038/s41467-020-15562-9',
'article-title': 'Characterization of spike glycoprotein of SARS-CoV-2 on virus entry and '
'its immune cross-reactivity with SARS-CoV',
'volume': '11',
'author': 'Ou',
'year': '2020',
'journal-title': 'Nat. Commun.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib34',
'doi-asserted-by': 'crossref',
'first-page': 'eabf7872',
'DOI': '10.1126/scitranslmed.abf7872',
'article-title': 'SARS-CoV-2 infection of human iPSC-derived cardiac cells reflects '
'cytopathic features in hearts of patients with COVID-19',
'volume': '13',
'author': 'Perez-Bermejo',
'year': '2021',
'journal-title': 'Sci. Transl. Med.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib35',
'doi-asserted-by': 'crossref',
'first-page': '2586',
'DOI': '10.1093/eurheartj/ehw160',
'article-title': 'ALPK3-deficient cardiomyocytes generated from patient-derived induced '
'pluripotent stem cells and mutant human embryonic stem cells display '
'abnormal calcium handling and establish that ALPK3 deficiency underlies '
'familial cardiomyopathy',
'volume': '37',
'author': 'Phelan',
'year': '2016',
'journal-title': 'Eur. Heart J.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib36',
'doi-asserted-by': 'crossref',
'first-page': '107940',
'DOI': '10.1016/j.celrep.2020.107940',
'article-title': 'Remdesivir inhibits SARS-CoV-2 in human lung cells and chimeric '
'SARS-CoV expressing the SARS-CoV-2 RNA polymerase in mice',
'volume': '32',
'author': 'Pruijssers',
'year': '2020',
'journal-title': 'Cell Rep.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib37',
'doi-asserted-by': 'crossref',
'first-page': '1265',
'DOI': '10.1001/jamacardio.2020.3557',
'article-title': 'Outcomes of cardiovascular magnetic resonance imaging in patients '
'recently recovered from coronavirus disease 2019 (COVID-19)',
'volume': '5',
'author': 'Puntmann',
'year': '2020',
'journal-title': 'JAMA Cardiol.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib38',
'doi-asserted-by': 'crossref',
'first-page': '135',
'DOI': '10.1016/j.bbrc.2020.03.044',
'article-title': 'Single cell RNA sequencing of 13 human tissues identify cell types and '
'receptors of human coronaviruses',
'volume': '526',
'author': 'Qi',
'year': '2020',
'journal-title': 'Biochem. Biophys. Res. Commun.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib39',
'doi-asserted-by': 'crossref',
'first-page': '88',
'DOI': '10.1016/j.yjmcc.2020.08.009',
'article-title': 'Genes encoding ACE2, TMPRSS2 and related proteins mediating SARS-CoV-2 '
'viral entry are upregulated with age in human cardiomyocytes',
'volume': '147',
'author': 'Robinson',
'year': '2020',
'journal-title': 'J.\xa0Mol. Cell. Cardiol.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib40',
'doi-asserted-by': 'crossref',
'first-page': '139',
'DOI': '10.1093/bioinformatics/btp616',
'article-title': 'edgeR: a Bioconductor package for differential expression analysis of '
'digital gene expression data',
'volume': '26',
'author': 'Robinson',
'year': '2010',
'journal-title': 'Bioinformatics'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib41',
'doi-asserted-by': 'crossref',
'first-page': '13638',
'DOI': '10.1038/s41598-021-92940-3',
'article-title': 'The RNA sensor MDA5 detects SARS-CoV-2 infection',
'volume': '11',
'author': 'Sampaio',
'year': '2021',
'journal-title': 'Sci. Rep.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib42',
'article-title': 'SARS-CoV-2 direct cardiac damage through spike-mediated cardiomyocyte '
'fusion',
'author': 'Schneider',
'year': '2020',
'journal-title': 'Research Square'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib43',
'doi-asserted-by': 'crossref',
'first-page': '11727',
'DOI': '10.1073/pnas.2003138117',
'article-title': 'Cell entry mechanisms of SARS-CoV-2',
'volume': '117',
'author': 'Shang',
'year': '2020',
'journal-title': 'Proc. Natl. Acad. Sci. USA'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib44',
'first-page': '224',
'article-title': 'Myocardial injury is associated with higher mortality in patients with '
'coronavirus disease 2019: a meta-analysis',
'volume': '17',
'author': 'Shao',
'year': '2020',
'journal-title': 'J.\xa0Geriatr. Cardiol.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib45',
'doi-asserted-by': 'crossref',
'first-page': '100052',
'DOI': '10.1016/j.xcrm.2020.100052',
'article-title': 'Human iPSC-derived cardiomyocytes are susceptible to SARS-CoV-2 '
'infection',
'volume': '1',
'author': 'Sharma',
'year': '2020',
'journal-title': 'Cell Rep. Med.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib46',
'doi-asserted-by': 'crossref',
'first-page': 'e1009800',
'DOI': '10.1371/journal.ppat.1009800',
'article-title': 'SARS-CoV-2 suppresses IFNbeta production mediated by NSP1, 5, 6, 15, '
'ORF6 and ORF7b but does not suppress the effects of added interferon',
'volume': '17',
'author': 'Shemesh',
'year': '2021',
'journal-title': 'PLoS Pathog.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib47',
'doi-asserted-by': 'crossref',
'first-page': '802',
'DOI': '10.1001/jamacardio.2020.0950',
'article-title': 'Association of cardiac injury with mortality in hospitalized patients '
'with COVID-19 in Wuhan, China',
'volume': '5',
'author': 'Shi',
'year': '2020',
'journal-title': 'JAMA Cardiol.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib48',
'doi-asserted-by': 'crossref',
'first-page': '681',
'DOI': '10.1038/s41591-020-0868-6',
'article-title': 'SARS-CoV-2 entry factors are highly expressed in nasal epithelial cells '
'together with innate immune genes',
'volume': '26',
'author': 'Sungnak',
'year': '2020',
'journal-title': 'Nat. Med.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib49',
'doi-asserted-by': 'crossref',
'first-page': '437',
'DOI': '10.1016/j.stemcr.2021.02.005',
'article-title': 'Revealing tissue-specific SARS-CoV-2 infection and host responses using '
'human stem cell-derived lung and cerebral organoids',
'volume': '16',
'author': 'Tiwari',
'year': '2021',
'journal-title': 'Stem Cell Rep.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib50',
'doi-asserted-by': 'crossref',
'first-page': '127',
'DOI': '10.1016/j.stem.2012.09.013',
'article-title': 'Distinct metabolic flow enables large-scale purification of mouse and '
'human pluripotent stem cell-derived cardiomyocytes',
'volume': '12',
'author': 'Tohyama',
'year': '2013',
'journal-title': 'Cell Stem Cell'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib51',
'doi-asserted-by': 'crossref',
'first-page': '1260419',
'DOI': '10.1126/science.1260419',
'article-title': 'Proteomics. Tissue-based map of the human proteome',
'volume': '347',
'author': 'Uhlén',
'year': '2015',
'journal-title': 'Science'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib52',
'doi-asserted-by': 'crossref',
'first-page': '24',
'DOI': '10.1038/s43856-021-00025-z',
'article-title': 'Characterization of SARS-CoV-2 and host entry factors distribution in a '
'COVID-19 autopsy series',
'volume': '1',
'author': 'Wang',
'year': '2021',
'journal-title': 'Commun. Med.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib53',
'doi-asserted-by': 'crossref',
'first-page': '926',
'DOI': '10.1038/s42003-021-02453-y',
'article-title': 'Human embryonic stem cell-derived cardiomyocyte platform screens '
'inhibitors of SARS-CoV-2 infection',
'volume': '4',
'author': 'Williams',
'year': '2021',
'journal-title': 'Commun. Biol.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib54',
'doi-asserted-by': 'crossref',
'first-page': '108234',
'DOI': '10.1016/j.celrep.2020.108234',
'article-title': 'Evasion of type I interferon by SARS-CoV-2',
'volume': '33',
'author': 'Xia',
'year': '2020',
'journal-title': 'Cell Rep.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib55',
'doi-asserted-by': 'crossref',
'first-page': '4',
'DOI': '10.1186/s41065-020-00168-4',
'article-title': 'Mediators of SARS-CoV-2 entry are preferentially enriched in '
'cardiomyocytes',
'volume': '158',
'author': 'Yang',
'year': '2021',
'journal-title': 'Hereditas'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib56',
'article-title': 'The FDA-approved drug Alectinib compromises SARS-CoV-2 nucleocapsid '
'phosphorylation and inhibits viral infection in\xa0vitro',
'author': 'Yaron',
'year': '2020',
'journal-title': 'bioRxiv'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib57',
'doi-asserted-by': 'crossref',
'first-page': '905',
'DOI': '10.1016/j.stem.2020.10.004',
'article-title': 'Three-dimensional human alveolar stem cell culture models reveal '
'infection response to SARS-CoV-2',
'volume': '27',
'author': 'Youk',
'year': '2020',
'journal-title': 'Cell Stem Cell'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib58',
'doi-asserted-by': 'crossref',
'first-page': 'eabd4570',
'DOI': '10.1126/science.abd4570',
'article-title': 'Inborn errors of type I IFN immunity in patients with life-threatening '
'COVID-19',
'volume': '370',
'author': 'Zhang',
'year': '2020',
'journal-title': 'Science'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib59',
'doi-asserted-by': 'crossref',
'first-page': 'E6',
'DOI': '10.1038/s41586-020-2951-z',
'article-title': 'Addendum: a pneumonia outbreak associated with a new coronavirus of '
'probable bat origin',
'volume': '588',
'author': 'Zhou',
'year': '2020',
'journal-title': 'Nature'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib60',
'doi-asserted-by': 'crossref',
'first-page': '1523',
'DOI': '10.1038/s41467-019-09234-6',
'article-title': 'Metascape provides a biologist-oriented resource for the analysis of '
'systems-level datasets',
'volume': '10',
'author': 'Zhou',
'year': '2019',
'journal-title': 'Nat. Commun.'},
{ 'key': '10.1016/j.stemcr.2023.05.007_bib61',
'doi-asserted-by': 'crossref',
'first-page': '185',
'DOI': '10.1007/s11684-020-0754-0',
'article-title': 'Single-cell RNA-seq data analysis on the receptor ACE2 expression '
'reveals the potential risk of different human organs vulnerable to '
'2019-nCoV infection',
'volume': '14',
'author': 'Zou',
'year': '2020',
'journal-title': 'Front. Med.'}],
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