Single-cell meta-analysis of SARS-CoV-2 entry genes across tissues and demographics.


Journal

Nature medicine
ISSN: 1546-170X
Titre abrégé: Nat Med
Pays: United States
ID NLM: 9502015

Informations de publication

Date de publication:
03 2021
Historique:
received: 16 04 2020
accepted: 23 12 2020
pubmed: 4 3 2021
medline: 27 3 2021
entrez: 3 3 2021
Statut: ppublish

Résumé

Angiotensin-converting enzyme 2 (ACE2) and accessory proteases (TMPRSS2 and CTSL) are needed for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) cellular entry, and their expression may shed light on viral tropism and impact across the body. We assessed the cell-type-specific expression of ACE2, TMPRSS2 and CTSL across 107 single-cell RNA-sequencing studies from different tissues. ACE2, TMPRSS2 and CTSL are coexpressed in specific subsets of respiratory epithelial cells in the nasal passages, airways and alveoli, and in cells from other organs associated with coronavirus disease 2019 (COVID-19) transmission or pathology. We performed a meta-analysis of 31 lung single-cell RNA-sequencing studies with 1,320,896 cells from 377 nasal, airway and lung parenchyma samples from 228 individuals. This revealed cell-type-specific associations of age, sex and smoking with expression levels of ACE2, TMPRSS2 and CTSL. Expression of entry factors increased with age and in males, including in airway secretory cells and alveolar type 2 cells. Expression programs shared by ACE2

Identifiants

pubmed: 33654293
doi: 10.1038/s41591-020-01227-z
pii: 10.1038/s41591-020-01227-z
pmc: PMC9469728
mid: NIHMS1757283
doi:

Substances chimiques

ACE2 protein, human EC 3.4.17.23
Angiotensin-Converting Enzyme 2 EC 3.4.17.23
Serine Endopeptidases EC 3.4.21.-
TMPRSS2 protein, human EC 3.4.21.-
CTSL protein, human EC 3.4.22.15
Cathepsin L EC 3.4.22.15

Types de publication

Journal Article Meta-Analysis Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

546-559

Subventions

Organisme : NHLBI NIH HHS
ID : K08 HL130595
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL127349
Pays : United States
Organisme : NIDDK NIH HHS
ID : P30 DK043351
Pays : United States
Organisme : Medical Research Council
ID : MR/S035907/1
Pays : United Kingdom
Organisme : NHLBI NIH HHS
ID : U01 HL148867
Pays : United States
Organisme : Medical Research Council
ID : MR/S035826/1
Pays : United Kingdom
Organisme : Medical Research Council
ID : MR/P009581/1
Pays : United Kingdom
Organisme : NHLBI NIH HHS
ID : R01 HL130938
Pays : United States
Organisme : NHLBI NIH HHS
ID : U01 HL145567
Pays : United States
Organisme : NHLBI NIH HHS
ID : U01 HL145550
Pays : United States
Organisme : NHLBI NIH HHS
ID : U01 HL148861
Pays : United States
Organisme : NHLBI NIH HHS
ID : F32 HL149290
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL141380
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL119215
Pays : United States
Organisme : NHLBI NIH HHS
ID : U01 HL148856
Pays : United States
Organisme : NIDDK NIH HHS
ID : RC2 DK114784
Pays : United States
Organisme : NIAID NIH HHS
ID : U19 AI116482
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL145372
Pays : United States
Organisme : NHLBI NIH HHS
ID : U01 HL122700
Pays : United States
Organisme : NIA NIH HHS
ID : P01 AG049665
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM007753
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK117263
Pays : United States
Organisme : NHLBI NIH HHS
ID : K08 HL146943
Pays : United States
Organisme : NIAID NIH HHS
ID : U19 AI135964
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL080494
Pays : United States
Organisme : NICHD NIH HHS
ID : R24 HD000836
Pays : United States
Organisme : NHLBI NIH HHS
ID : R35 HL140039
Pays : United States
Organisme : Medical Research Council
ID : MR/S005579/1
Pays : United Kingdom
Organisme : NHLBI NIH HHS
ID : R01 HL146519
Pays : United States
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : NIAID NIH HHS
ID : U24 AI118672
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL133153
Pays : United States
Organisme : British Heart Foundation
ID : SP/19/1/34461
Pays : United Kingdom
Organisme : NHLBI NIH HHS
ID : R01 HL146557
Pays : United States
Organisme : NIAID NIH HHS
ID : U19 AI142784
Pays : United States
Organisme : NHLBI NIH HHS
ID : R56 HL135124
Pays : United States

Investigateurs

Gail H Deutsch (GH)
Jennifer Dutra (J)
Kyle J Gaulton (KJ)
Jeanne Holden-Wiltse (J)
Heidie L Huyck (HL)
Thomas J Mariani (TJ)
Ravi S Misra (RS)
Cory Poole (C)
Sebastian Preissl (S)
Gloria S Pryhuber (GS)
Lisa Rogers (L)
Xin Sun (X)
Allen Wang (A)
Jeffrey A Whitsett (JA)
Yan Xu (Y)
Jehan Alladina (J)
Nicholas E Banovich (NE)
Pascal Barbry (P)
Jennifer E Beane (JE)
Roby P Bhattacharyya (RP)
Katharine E Black (KE)
Alvis Brazma (A)
Joshua D Campbell (JD)
Josalyn L Cho (JL)
Joseph Collin (J)
Christian Conrad (C)
Kitty de Jong (K)
Tushar Desai (T)
Diane Z Ding (DZ)
Oliver Eickelberg (O)
Roland Eils (R)
Patrick T Ellinor (PT)
Alen Faiz (A)
Christine S Falk (CS)
Michael Farzan (M)
Andrew Gellman (A)
Gad Getz (G)
Ian A Glass (IA)
Anna Greka (A)
Muzlifah Haniffa (M)
Lida P Hariri (LP)
Mark W Hennon (MW)
Peter Horvath (P)
Norbert Hübner (N)
Deborah T Hung (DT)
Heidie L Huyck (HL)
William J Janssen (WJ)
Dejan Juric (D)
Naftali Kaminski (N)
Melanie Koenigshoff (M)
Gerard H Koppelman (GH)
Mark A Krasnow (MA)
Jonathan A Kropski (JA)
Malte Kuhnemund (M)
Robert Lafyatis (R)
Majlinda Lako (M)
Eric S Lander (ES)
Haeock Lee (H)
Marc E Lenburg (ME)
Charles-Hugo Marquette (CH)
Ross J Metzger (RJ)
Sten Linnarsson (S)
Gang Liu (G)
Yuk Ming Dennis Lo (YMD)
Joakim Lundeberg (J)
John C Marioni (JC)
Sarah A Mazzilli (SA)
Benjamin D Medoff (BD)
Kerstin B Meyer (KB)
Zhichao Miao (Z)
Alexander V Misharin (AV)
Martijn C Nawijn (MC)
Marko Z Nikolić (MZ)
Michela Noseda (M)
Jose Ordovas-Montanes (J)
Gavin Y Oudit (GY)
Dana Pe'er (D)
Joseph E Powell (JE)
Stephen R Quake (SR)
Jayaraj Rajagopal (J)
Purushothama Rao Tata (PR)
Emma L Rawlins (EL)
Aviv Regev (A)
Mary E Reid (ME)
Paul A Reyfman (PA)
Kimberly M Rieger-Christ (KM)
Mauricio Rojas (M)
Orit Rozenblatt-Rosen (O)
Kourosh Saeb-Parsy (K)
Christos Samakovlis (C)
Joshua R Sanes (JR)
Herbert B Schiller (HB)
Joachim L Schultze (JL)
Roland F Schwarz (RF)
Ayellet V Segre (AV)
Max A Seibold (MA)
Christine E Seidman (CE)
Jon G Seidman (JG)
Alex K Shalek (AK)
Douglas P Shepherd (DP)
Rahul Sinha (R)
Jason R Spence (JR)
Avrum Spira (A)
Xin Sun (X)
Erik Sundström (E)
Sarah A Teichmann (SA)
Fabian J Theis (FJ)
Alexander M Tsankov (AM)
Ludovic Vallier (L)
Maarten van den Berge (M)
Tave A Van Zyl (TA)
Alexandra-Chloé Villani (AC)
Astrid Weins (A)
Ramnik J Xavier (RJ)
Ali Önder Yildirim (AÖ)
Laure-Emmanuelle Zaragosi (LE)
Darin Zerti (D)
Hongbo Zhang (H)
Kun Zhang (K)
Xiaohui Zhang (X)

Références

Wang, D. et al. Clinical characteristics of 138 hospitalized patients with 2019 novel coronavirus–infected pneumonia in Wuhan, China. JAMA 323, 1061–1069 (2020).
pubmed: 32031570 pmcid: 7042881 doi: 10.1001/jama.2020.1585
Huang, C. et al. Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China. Lancet 395, 497–506 (2020).
pubmed: 31986264 pmcid: 7159299 doi: 10.1016/S0140-6736(20)30183-5
Chen, N. et al. Epidemiological and clinical characteristics of 99 cases of 2019 novel coronavirus pneumonia in Wuhan, China: a descriptive study. Lancet 395, 507–513 (2020).
pubmed: 32007143 pmcid: 7135076 doi: 10.1016/S0140-6736(20)30211-7
Wang, W. et al. Detection of SARS-CoV-2 in different types of clinical specimens. JAMA https://doi.org/10.1001/jama.2020.3786 (2020).
Jia, H. P. et al. ACE2 receptor expression and severe acute respiratory syndrome coronavirus infection depend on differentiation of human airway epithelia. J. Virol. 79, 14614–14621 (2005).
pubmed: 16282461 pmcid: 1287568 doi: 10.1128/JVI.79.23.14614-14621.2005
Hou, Y. J. et al. SARS-CoV-2 reverse genetics reveals a variable infection gradient in the respiratory tract. Cell 182, 429–446 (2020).
pubmed: 32526206 pmcid: 7250779 doi: 10.1016/j.cell.2020.05.042
McCray, P. B. Jr et al. Lethal infection of K18-hACE2 mice infected with severe acute respiratory syndrome coronavirus. J. Virol. 81, 813–821 (2007).
pubmed: 17079315 doi: 10.1128/JVI.02012-06
Walls, A. C. et al. Structure, function and antigenicity of the SARS-CoV-2 spike glycoprotein. Cell 181, 281–292 (2020).
pubmed: 32155444 pmcid: 7102599 doi: 10.1016/j.cell.2020.02.058
Perez-Saez, J. et al. Serology-informed estimates of SARS-CoV-2 infection fatality risk in Geneva, Switzerland. Lancet Infect. Dis. https://doi.org/10.1016/S1473-3099(20)30584-3 (2020).
Zhou, F. et al. Clinical course and risk factors for mortality of adult inpatients with COVID-19 in Wuhan, China: a retrospective cohort study. Lancet 395, 1054–1062 (2020).
pubmed: 32171076 pmcid: 7270627 doi: 10.1016/S0140-6736(20)30566-3
Ludvigsson, J. F. Systematic review of COVID‐19 in children shows milder cases and a better prognosis than adults. Acta Paediatr. 109, 1088–1095 (2020).
pubmed: 32202343 doi: 10.1111/apa.15270
Guo, F. R. Smoking links to the severity of COVID‐19: an update of a meta‐analysis. J. Med. Virol. 92, 2304–2305 (2020).
Sungnak, W. et al. SARS-CoV-2 entry factors are highly expressed in nasal epithelial cells together with innate immune genes. Nat. Med. 26, 681–687 (2020).
pubmed: 32327758 pmcid: 8637938 doi: 10.1038/s41591-020-0868-6
Ziegler, C. G. K. et al. SARS-CoV-2 receptor ACE2 is an interferon-stimulated gene in human airway epithelial cells and is detected in specific cell subsets across tissues. Cell 181, 1016–1035 (2020).
pubmed: 32413319 pmcid: 7252096 doi: 10.1016/j.cell.2020.04.035
Qi, F., Qian, S., Zhang, S. & Zhang, Z. Single-cell RNA sequencing of 13 human tissues identify cell types and receptors of human coronaviruses. Biochem. Biophys. Res. Commun. 526, 135–140 (2020).
pubmed: 32199615 pmcid: 7156119 doi: 10.1016/j.bbrc.2020.03.044
Lukassen, S. et al. SARS-CoV-2 receptor ACE2 and TMPRSS2 are primarily expressed in bronchial transient secretory cells. EMBO J. https://doi.org/10.15252/embj.20105114 (2020).
Zhang, H. et al. Specific ACE2 expression in small intestinal enterocytes may cause gastrointestinal symptoms and injury after 2019-nCoV infection. Int. J. Infect. Dis. 96, 19–24 (2020).
pubmed: 32311451 pmcid: 7165079 doi: 10.1016/j.ijid.2020.04.027
Ordovas-Montanes, J. et al. Allergic inflammatory memory in human respiratory epithelial progenitor cells. Nature 560, 649–654 (2018).
pubmed: 30135581 pmcid: 6133715 doi: 10.1038/s41586-018-0449-8
Sos, B. C. et al. Characterization of chromatin accessibility with a transposome hypersensitive sites sequencing assay. Genome Biol. 17, 20 (2016).
pubmed: 26846207 pmcid: 4743176 doi: 10.1186/s13059-016-0882-7
Emery, B. et al. Myelin gene regulatory factor is a critical transcriptional regulator required for CNS myelination. Cell 138, 172–185 (2009).
pubmed: 19596243 pmcid: 2757090 doi: 10.1016/j.cell.2009.04.031
Vento-Tormo, R. et al. Single-cell reconstruction of the early maternal–fetal interface in humans. Nature 563, 347–353 (2018).
pubmed: 30429548 doi: 10.1038/s41586-018-0698-6
Suryawanshi, H. et al. A single-cell survey of the human first-trimester placenta and decidua. Sci. Adv. 4, eaau4788 (2018).
pubmed: 30402542 pmcid: 6209386 doi: 10.1126/sciadv.aau4788
Tsang, J. C. H. et al. Integrative single-cell and cell-free plasma RNA transcriptomics elucidates placental cellular dynamics. Proc. Natl Acad. Sci. USA 114, E7786–E7795 (2017).
pubmed: 28830992 pmcid: 5604038 doi: 10.1073/pnas.1710470114
Pérez-Silva, J. G., Español, Y., Velasco, G. & Quesada, V. The Degradome database: expanding roles of mammalian proteases in life and disease. Nucleic Acids Res. 44, D351–D355 (2016).
pubmed: 26553809 doi: 10.1093/nar/gkv1201
Coutard, B. et al. The spike glycoprotein of the new coronavirus 2019-nCoV contains a furin-like cleavage site absent in CoV of the same clade. Antiviral Res. 176, 104742 (2020).
pubmed: 32057769 pmcid: 7114094 doi: 10.1016/j.antiviral.2020.104742
Millet, J. K. & Whittaker, G. R. Physiological and molecular triggers for SARS-CoV membrane fusion and entry into host cells. Virology 517, 3–8 (2018).
pubmed: 29275820 doi: 10.1016/j.virol.2017.12.015
Seidah, N. G. & Prat, A. The biology and therapeutic targeting of the proprotein convertases. Nat. Rev. Drug Discov. 11, 367–383 (2012).
pubmed: 22679642 doi: 10.1038/nrd3699
Cai, H. Sex difference and smoking predisposition in patients with COVID-19. Lancet Respir. Med. 8, e20 (2020).
pubmed: 32171067 pmcid: 7103991 doi: 10.1016/S2213-2600(20)30117-X
Goldfarbmuren, K. C. et al. Dissecting the cellular specificity of smoking effects and reconstructing lineages in the human airway epithelium. Nat. Commun. 11, 2485 (2020).
pubmed: 32427931 pmcid: 7237663 doi: 10.1038/s41467-020-16239-z
Duclos, G. E. et al. Characterizing smoking-induced transcriptional heterogeneity in the human bronchial epithelium at single-cell resolution. Sci. Adv. 5, eaaw3413 (2019).
pubmed: 31844660 pmcid: 6905872 doi: 10.1126/sciadv.aaw3413
Vieira Braga, F. A. et al. A cellular census of human lungs identifies novel cell states in health and in asthma. Nat. Med. 25, 1153–1163 (2019).
pubmed: 31209336 doi: 10.1038/s41591-019-0468-5
Reyfman, P. A. et al. Single-cell transcriptomic analysis of human lung provides insights into the pathobiology of pulmonary fibrosis. Am. J. Respir. Crit. Care Med. 199, 1517–1536 (2019).
pubmed: 30554520 pmcid: 6580683 doi: 10.1164/rccm.201712-2410OC
Madissoon, E. et al. scRNA-seq assessment of the human lung, spleen and esophagus tissue stability after cold preservation. Genome Biol. 21, 1 (2019).
pubmed: 31892341 pmcid: 6937944 doi: 10.1186/s13059-019-1906-x
Miller, A. J. et al. In vitro and in vivo development of the human airway at single-cell resolution. Dev. Cell 53, 117–128 (2020).
pubmed: 32109386 pmcid: 7396815 doi: 10.1016/j.devcel.2020.01.033
Adams, T. S. et al. Single-cell RNA-seq reveals ectopic and aberrant lung-resident cell populations in idiopathic pulmonary fibrosis. Sci. Adv. 6, eaba1983 (2020).
Habermann, A. C. et al. Single-cell RNA sequencing reveals profibrotic roles of distinct epithelial and mesenchymal lineages in pulmonary fibrosis. Sci. Adv. 6, eaba1972 (2020).
pubmed: 32832598 pmcid: 7439444 doi: 10.1126/sciadv.aba1972
Deprez, M. et al. A single-cell atlas of the human healthy airways. Am. J. Respir. Crit. Care Med. https://doi.org/10.1164/rccm.201911-2199OC (2020).
Morse, C. et al. Proliferating SPP1/MERTK-expressing macrophages in idiopathic pulmonary fibrosis. Eur. Respir. J. 54, 1802441 (2019).
pubmed: 31221805 pmcid: 8025672 doi: 10.1183/13993003.02441-2018
Travaglini, K. J., Nabhan, A. N., Penland, L. & Sinha, R. A molecular cell atlas of the human lung from single-cell RNA sequencing. Nature 587, 619–625 (2020).
pubmed: 33208946 pmcid: 7704697 doi: 10.1038/s41586-020-2922-4
Mayr, C. H. et al. Integrated single-cell analysis of human lung fibrosis resolves cellular origins of predictive protein signatures in body fluids. SSRN https://doi.org/10.2139/ssrn.3538700 (2020).
Beane, J. E. et al. Molecular subtyping reveals immune alterations associated with progression of bronchial premalignant lesions. Nat. Commun. 10, 1856 (2019).
pubmed: 31015447 pmcid: 6478943 doi: 10.1038/s41467-019-09834-2
Chan, C.-M. et al. Carcinoembryonic antigen-related cell adhesion molecule 5 is an important surface attachment factor that facilitates entry of middle east respiratory syndrome coronavirus. J.Virol. 90, 9114–9127 (2016).
pubmed: 27489282 pmcid: 5044831 doi: 10.1128/JVI.01133-16
Wahl, S. M. et al. Secretory leukocyte protease inhibitor in mucosal fluids inhibits HIV-1. Oral Dis. 3, S64–S69 (1997).
pubmed: 9456660 doi: 10.1111/j.1601-0825.1997.tb00377.x
Turula, H. & Wobus, C. The role of the polymeric immunoglobulin receptor and secretory immunoglobulins during mucosal infection and immunity. Viruses 10, 237 (2018).
pmcid: 5977230 doi: 10.3390/v10050237
Burkhardt, A. M. et al. CXCL17 is a mucosal chemokine elevated in idiopathic pulmonary fibrosis that exhibits broad antimicrobial activity. J. Immunol. 188, 6399–6406 (2012).
pubmed: 22611239 doi: 10.4049/jimmunol.1102903
Debbabi, H. et al. Primary type II alveolar epithelial cells present microbial antigens to antigen-specific CD4
pubmed: 15833765 doi: 10.1152/ajplung.00004.2005
Yue, Y. et al. SARS-Coronavirus open reading frame-3a drives multimodal necrotic cell death. Cell Death Dis. 9, 904 (2018).
pubmed: 30185776 pmcid: 6125346 doi: 10.1038/s41419-018-0917-y
Burkard, C. et al. Coronavirus cell entry occurs through the endo-/lysosomal pathway in a proteolysis-dependent manner. PLoS Pathog. 10, e1004502 (2014).
pubmed: 25375324 pmcid: 4223067 doi: 10.1371/journal.ppat.1004502
Wishart, D. S. et al. DrugBank 5.0: a major update to the DrugBank database for 2018. Nucleic Acids Res. 46, D1074–D1082 (2018).
pubmed: 29126136 doi: 10.1093/nar/gkx1037
Gordon, D. E. et al. A SARS-CoV-2 protein interaction map reveals targets for drug repurposing. Nature 583, 459–468 (2020).
pubmed: 32353859 pmcid: 7431030 doi: 10.1038/s41586-020-2286-9
Law, C. W., Chen, Y., Shi, W. & Smyth, G. K. voom: precision weights unlock linear model analysis tools for RNA-seq read counts. Genome Biol. 15, R29 (2014).
pubmed: 24485249 pmcid: 4053721 doi: 10.1186/gb-2014-15-2-r29
Luan, H. H. et al. GDF15 is an inflammation-induced central mediator of tissue tolerance. Cell 178, 1231–1244 (2019).
pubmed: 31402172 pmcid: 6863354 doi: 10.1016/j.cell.2019.07.033
Dhar, P. & McAuley, J. The role of the cell surface mucin MUC1 as a barrier to infection and regulator of inflammation. Front. Cell. Infect. Microbiol. 9, 117 (2019).
pubmed: 31069176 pmcid: 6491460 doi: 10.3389/fcimb.2019.00117
Efremova, M., Vento-Tormo, M., Teichmann, S. A. & Vento-Tormo, R. CellPhoneDB: inferring cell–cell communication from combined expression of multi-subunit ligand–receptor complexes. Nat. Protoc. 15, 1484–1506 (2020).
pubmed: 32103204 doi: 10.1038/s41596-020-0292-x
Bao, L. et al. The pathogenicity of SARS-CoV-2 in hACE2 transgenic mice. Nature 583, 830–833 (2020).
pubmed: 32380511 doi: 10.1038/s41586-020-2312-y
Montoro, D. T. et al. A revised airway epithelial hierarchy includes CFTR-expressing ionocytes. Nature 560, 319–324 (2018).
pubmed: 30069044 pmcid: 6295155 doi: 10.1038/s41586-018-0393-7
Smith, J. C. et al. Cigarette smoke exposure and inflammatory signaling increase the expression of the SARS-CoV-2 receptor ACE2 in the respiratory tract. Devel. Cell 53, 514–529.e3 (2020).
doi: 10.1016/j.devcel.2020.05.012
Booeshaghi, A. S. & Pachter, L. Decrease in ACE2 mRNA expression in aged mouse lung. Preprint at bioRxiv https://doi.org/10.1101/2020.04.02.021451 (2020).
Tucker Nathan, R. et al. Myocyte-specific upregulation of ACE2 in cardiovascular disease. Circulation 142, 708–710 (2020).
pubmed: 32795091 pmcid: 7424896
Hamming, I. et al. Tissue distribution of ACE2 protein, the functional receptor for SARS coronavirus. A first step in understanding SARS pathogenesis. J. Pathol. 203, 631–637 (2004).
pubmed: 15141377 pmcid: 7167720 doi: 10.1002/path.1570
Zhao, Y. et al. Single-cell RNA expression profiling of ACE2, the receptor of SARS-CoV-2. Am. J. Respir. Crit. Care Med. 202, 756–759 (2020).
pubmed: 32663409 pmcid: 7462411 doi: 10.1164/rccm.202001-0179LE
Venkatakrishnan, A. J. et al. Knowledge synthesis of 100 million biomedical documents augments the deep expression profiling of coronavirus receptors. eLife 9, e58040 (2020).
pubmed: 32463365 pmcid: 7371427 doi: 10.7554/eLife.58040
Mao, L. et al. Neurological manifestations of hospitalized patients with COVID-19 in Wuhan, China: a retrospective case series study. JAMA Neurol. 77, 683–690 (2020).
Poyiadji, N. et al. COVID-19-associated acute hemorrhagic necrotizing encephalopathy: CT and MRI features. Radiology 296, E119–E120 (2020).
Helms, J., Kremer, S. & Meziani, F. More on neurologic features in severe SARS-CoV-2 infection. N. Engl. J. Med. 382, e110 (2020).
pubmed: 32453517 doi: 10.1056/NEJMc2008597
Toscano, G. et al. Guillain–Barré syndrome associated with SARS-CoV-2. N. Engl. J. Med. 382, 2574–2576 (2020).
pubmed: 32302082 doi: 10.1056/NEJMc2009191
Del Valle, D. M. et al. An inflammatory cytokine signature predicts COVID-19 severity and survival. Nat. Med. https://doi.org/10.1038/s41591-020-1051-9 (2020).
Liao, M. et al. Single-cell landscape of bronchoalveolar immune cells in patients with COVID-19. Nat. Med. 26, 842–844 (2020).
pubmed: 32398875 doi: 10.1038/s41591-020-0901-9
McInnes, L. et al. UMAP: Uniform Manifold Approximation and Projection. J. Open Source Softw. 3, 861 (2018).
doi: 10.21105/joss.00861
Korsunsky, I. et al. Fast, sensitive and accurate integration of single-cell data with Harmony. Nat. Methods 16, 1289–1296 (2019).
pubmed: 31740819 pmcid: 6884693 doi: 10.1038/s41592-019-0619-0
Seabold, S. & Perktold, J. Statsmodels: econometric and statistical modeling with python. In Proceedings of the 9th Python in Science Conference 57–61 (Austin, 2010).
Wolf, F. A., Angerer, P. & Theis, F. J. SCANPY: large-scale single-cell gene expression data analysis. Genome Biol. 19, 15 (2018).
pubmed: 29409532 pmcid: 5802054 doi: 10.1186/s13059-017-1382-0
West, B. T., Welch, K. B. & Galecki, A. T. Linear Mixed Models: a Practical Guide Using Statistical Software 2nd edn. (CRC Press, 2014).
Stuart, T. et al. Comprehensive integration of single-cell data. Cell 177, 1888–1902 (2019).
pubmed: 31178118 pmcid: 6687398 doi: 10.1016/j.cell.2019.05.031
Cusanovich, D. A. et al. Multiplex single-cell profiling of chromatin accessibility by combinatorial cellular indexing. Science 348, 910–914 (2015).
pubmed: 25953818 pmcid: 4836442 doi: 10.1126/science.aab1601
Schep, A. N., Wu, B., Buenrostro, J. D. & Greenleaf, W. J. chromVAR: inferring transcription-factor-associated accessibility from single-cell epigenomic data. Nat. Methods 14, 975–978 (2017).
pubmed: 28825706 pmcid: 5623146 doi: 10.1038/nmeth.4401
Fornes, O. et al. JASPAR 2020: update of the open-access database of transcription factor binding profiles. Nucleic Acids Res. 48, D87–D92 (2020).
pubmed: 31701148
Pedregosa, F. et al. Scikit-learn: machine learning in Python. J. Mach. Learn. Res. 12, 2825–2830 (2011).
Breiman, L., Friedman, J., Stone, C. J. & Olshen, R. A. Classification and Regression Trees (CRC press, 1984).
Jacomy, M., Venturini, T., Heymann, S. & Bastian, M. ForceAtlas2, a continuous graph layout algorithm for handy network visualization designed for the Gephi software. PLoS ONE 9, e98679 (2014).
pubmed: 24914678 pmcid: 4051631 doi: 10.1371/journal.pone.0098679
Raudvere, U. et al. g:Profiler: a web server for functional enrichment analysis and conversions of gene lists. Nucleic Acids Res. 47, W191–W198 (2019).
pubmed: 31066453 pmcid: 6602461 doi: 10.1093/nar/gkz369
Finak, G. et al. MAST: a flexible statistical framework for assessing transcriptional changes and characterizing heterogeneity in single-cell RNA sequencing data. Genome Biol. 16, 278 (2015).
pubmed: 26653891 pmcid: 4676162 doi: 10.1186/s13059-015-0844-5
Kuleshov, M. V. et al. Enrichr: a comprehensive gene set enrichment analysis web server. Nucleic Acids Res. 44, W90–W97 (2016).
pubmed: 27141961 pmcid: 4987924 doi: 10.1093/nar/gkw377
Jia, J. et al. Cholesterol metabolism promotes B cell positioning during immune pathogenesis of chronic obstructive pulmonary disease. EMBO Mol. Med. 10, e8349 (2018).
pubmed: 29674392 pmcid: 5938615 doi: 10.15252/emmm.201708349

Auteurs

Christoph Muus (C)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA. muus@broadinstitute.org.
John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA, USA. muus@broadinstitute.org.

Malte D Luecken (MD)

Institute of Computational Biology, Helmholtz Zentrum München, Neuherberg, Germany. malte.luecken@helmholtz-muenchen.de.

Gökcen Eraslan (G)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Lisa Sikkema (L)

Institute of Computational Biology, Helmholtz Zentrum München, Neuherberg, Germany.

Avinash Waghray (A)

Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA, USA.
Departments of Internal Medicine and Pediatrics, Pulmonary and Critical Care Unit, Massachusetts General Hospital, Boston, MA, USA.
Harvard Stem Cell Institute, Cambridge, MA, USA.

Graham Heimberg (G)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Yoshihiko Kobayashi (Y)

Department of Cell Biology, Duke University Medical School, Durham, NC, USA.

Eeshit Dhaval Vaishnav (ED)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA.

Ayshwarya Subramanian (A)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Christopher Smillie (C)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Karthik A Jagadeesh (KA)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Elizabeth Thu Duong (ET)

Division of Respiratory Medicine, Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Evgenij Fiskin (E)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Elena Torlai Triglia (E)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Meshal Ansari (M)

Comprehensive Pneumology Center (CPC)/Institute of Lung Biology and Disease (ILBD), Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), Munich, Germany.
Institute of Computational Biology, Helmholtz Zentrum München, Munich, Germany.

Peiwen Cai (P)

Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, USA.

Brian Lin (B)

Departments of Internal Medicine and Pediatrics, Pulmonary and Critical Care Unit, Massachusetts General Hospital, Boston, MA, USA.
Harvard Stem Cell Institute, Cambridge, MA, USA.
Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA, USA.

Justin Buchanan (J)

Center for Epigenomics, University of California San Diego School of Medicine, La Jolla, CA, USA.
Department of Cellular and Molecular Medicine, University of California San Diego School of Medicine, La Jolla, CA, USA.

Sijia Chen (S)

Division of Rheumatology, Inflammation, and Immunity, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.

Jian Shu (J)

Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Whitehead Institute for Biomedical Research, Cambridge, MA, USA.

Adam L Haber (AL)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Department of Environmental Health, Harvard T. H. Chan School of Public Health, Boston, MA, USA.

Hattie Chung (H)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Daniel T Montoro (DT)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Taylor Adams (T)

Pulmonary, Critical Care and Sleep Medicine, Yale University School of Medicine, New Haven, CT, USA.

Hananeh Aliee (H)

Institute of Computational Biology, Helmholtz Zentrum München, Munich, Germany.

Samuel J Allon (SJ)

Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Institute for Medical Engineering and Science & Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, USA.
Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, USA.

Zaneta Andrusivova (Z)

SciLifeLab, Department of Gene Technology, KTH Royal Institute of Technology, Stockolm, Sweden.

Ilias Angelidis (I)

Comprehensive Pneumology Center (CPC)/Institute of Lung Biology and Disease (ILBD), Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), Munich, Germany.

Orr Ashenberg (O)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Kevin Bassler (K)

Department for Genomics & Immunoregulation, LIMES-Institute, University of Bonn, Bonn, Germany.

Christophe Bécavin (C)

Université Côte d'Azur, CNRS, IPMC, Sophia-Antipolis, France.

Inbal Benhar (I)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Joseph Bergenstråhle (J)

SciLifeLab, Department of Gene Technology, KTH Royal Institute of Technology, Stockolm, Sweden.

Ludvig Bergenstråhle (L)

SciLifeLab, Department of Gene Technology, KTH Royal Institute of Technology, Stockolm, Sweden.

Liam Bolt (L)

Wellcome Sanger Institute, Hinxton, UK.

Emelie Braun (E)

Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden.

Linh T Bui (LT)

Translational Genomics Research Institute, Phoenix, AZ, USA.

Steven Callori (S)

Department of Medicine, Boston University School of Medicine, Boston, MA, USA.
Bioinformatics Program, Boston University School of Medicine, Boston, MA, USA.

Mark Chaffin (M)

Precision Cardiology Laboratory, The Broad Institute, Cambridge, MA, USA.

Evgeny Chichelnitskiy (E)

Institute of Transplant Immunology, Hannover Medical School, MHH, Hannover, Germany.
German Center for Infectious Diseases (DZIF), Braunschweig, Germany.

Joshua Chiou (J)

Biomedical Sciences Graduate Program, University of California, San Diego, La Jolla, CA, USA.

Thomas M Conlon (TM)

Comprehensive Pneumology Center (CPC)/Institute of Lung Biology and Disease (ILBD), Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), Munich, Germany.

Michael S Cuoco (MS)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Anna S E Cuomo (ASE)

European Molecular Biology Laboratory, European Bioinformatics Institute, Hinxton, UK.

Marie Deprez (M)

Université Côte d'Azur, CNRS, IPMC, Sophia-Antipolis, France.

Grant Duclos (G)

Boston University School of Medicine, Boston, MA, USA.

Denise Fine (D)

Boston University Medical Center, Boston, MA, USA.

David S Fischer (DS)

Institute of Computational Biology, Helmholtz Zentrum München, Munich, Germany.
TUM School of Life Sciences Weihenstephan, Technical University of Munich, Freising, Germany.

Shila Ghazanfar (S)

Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK.

Astrid Gillich (A)

Department of Biochemistry and Wall Center for Pulmonary Vascular Disease, Stanford, CA, USA.

Bruno Giotti (B)

Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, USA.

Joshua Gould (J)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Minzhe Guo (M)

Divisions of Pulmonary Biology; Perinatal Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA.

Austin J Gutierrez (AJ)

Translational Genomics Research Institute, Phoenix, AZ, USA.

Arun C Habermann (AC)

Division of Allergy, Pulmonary and Critical Care Medicine, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, USA.

Tyler Harvey (T)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Peng He (P)

Wellcome Sanger Institute, Hinxton, UK.

Xiaomeng Hou (X)

Center for Epigenomics, University of California-San Diego School of Medicine, La Jolla, CA, USA.
Department of Cellular and Molecular Medicine, University of California-San Diego School of Medicine, La Jolla, CA, USA.

Lijuan Hu (L)

Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden.

Yan Hu (Y)

Division of Pulmonary Sciences and Critical Care Medicine, School of Medicine, University of Colorado, Aurora, CO, USA.

Alok Jaiswal (A)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Lu Ji (L)

Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, SAR, China.

Peiyong Jiang (P)

Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, SAR, China.

Theodoros S Kapellos (TS)

Genomics and Immunoregulation, Life & Medical Sciences (LIMES) Institute, University of Bonn, Bonn, Germany.

Christin S Kuo (CS)

Division of Pulmonary Medicine, Department of Pediatrics, Stanford University, Stanford, CA, USA.

Ludvig Larsson (L)

SciLifeLab, Department of Gene Technology, KTH Royal Institute of Technology, Stockholm, Sweden.

Michael A Leney-Greene (MA)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Kyungtae Lim (K)

Gurdon Institute, University of Cambridge, Cambridge, UK.

Monika Litviňuková (M)

Cellular Genetics Programme, Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, UK.
Cardiovascular and Metabolic Sciences, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.

Leif S Ludwig (LS)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Division of Hematology/Oncology, Boston Children's Hospital and Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA.

Soeren Lukassen (S)

Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Berlin, Germany.
Berlin Institute of Health (BIH), Center for Digital Health, Berlin, Germany.

Wendy Luo (W)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Henrike Maatz (H)

Cardiovascular and Metabolic Sciences, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.

Elo Madissoon (E)

European Molecular Biology Laboratory-European Bioinformatics Institute, Wellcome Genome Campus, Hinxton, UK.
Wellcome Sanger Institute, Cellular Genetics Programme Wellcome Genome Campus, Hinxton, UK.

Lira Mamanova (L)

Wellcome Sanger Institute, Hinxton, UK.

Kasidet Manakongtreecheep (K)

Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Center for Cancer Research, Department of Medicine, Massachusetts General Hospital, Boston, MA, USA.
Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital, Charlestown, MA, USA.

Sylvie Leroy (S)

Pulmonology Department, Université Côte d'Azur, CHU Nice, Nice, France.
CNRS, Institut de Pharmacologie Moléculaire et Cellulaire, Sophia-Antipolis, France.

Christoph H Mayr (CH)

Helmholtz Zentrum München, Institute of Lung Biology and Disease, Group Systems Medicine of Chronic Lung Disease, Member of the German Center for Lung Research (DZL), Munich, Germany.

Ian M Mbano (IM)

Africa Health Research Institute, Durban, South Africa.
School of Laboratory Medicine and Medical Sciences, College of Health Sciences, University of Kwazulu Natal, Durban, South Africa.

Alexi M McAdams (AM)

Department of Ophthalmology, Harvard Medical School and Massachusetts Eye and Ear, Boston, MA, USA.

Ahmad N Nabhan (AN)

Department of Biochemistry and Wall Center for Pulmonary Vascular Disease, Stanford, CA, USA.

Sarah K Nyquist (SK)

Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, USA.
Computational and Systems Biology, CSAIL, Institute for Medical Engineering and Science & Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, USA.

Lolita Penland (L)

Department of Biochemistry and Wall Center for Pulmonary Vascular Disease, Stanford, CA, USA.

Olivier B Poirion (OB)

Center for Epigenomics, University of California-San Diego School of Medicine, La Jolla, CA, USA.
Department of Cellular and Molecular Medicine, University of California-San Diego School of Medicine, La Jolla, CA, USA.

Sergio Poli (S)

Pulmonary, Critical Care and Sleep Medicine, Yale University School of Medicine, New Haven, CT, USA.

CanCan Qi (C)

Department of Pediatric Pulmonology and Pediatric Allergology, Beatrix Children's Hospital, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
GRIAC Research Institute, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.

Rachel Queen (R)

Biosciences Institute, Faculty of Medical Sciences, Newcastle University, International Centre for Life, Newcastle upon Tyne, UK.

Daniel Reichart (D)

Department of Genetics, Harvard Medical School, Boston, MA, USA.
Department of Cardiology, University Heart & Vascular Center, University of Hamburg, Hamburg, Germany.

Ivan Rosas (I)

Pulmonary, Critical Care and Sleep Medicine, Yale University School of Medicine, New Haven, CT, USA.

Jonas C Schupp (JC)

Section of Pulmonary, Critical Care, and Sleep Medicine, Yale University School of Medicine, New Haven, CT, USA.

Conor V Shea (CV)

Division of Computational Biomedicine, Department of Medicine, Boston University School of Medicine, Boston, MA, USA.

Xingyi Shi (X)

Division of Computational Biomedicine, Department of Medicine, Boston University School of Medicine, Boston, MA, USA.
Bioinformatics Program, Boston University, Boston, MA, USA.

Rahul Sinha (R)

Institute for Stem Cell Biology and Regenerative Medicine, Stanford Medicine, Stanford, CA, USA.

Rene V Sit (RV)

Department of Biochemistry and Wall Center for Pulmonary Vascular Disease, Stanford, CA, USA.

Kamil Slowikowski (K)

Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Center for Cancer Research, Department of Medicine, Massachusetts General Hospital, Boston, MA, USA.
Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital, Charlestown, MA, USA.

Michal Slyper (M)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Neal P Smith (NP)

Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital, Boston, MA, USA.

Alex Sountoulidis (A)

Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Stockholm, Sweden.

Maximilian Strunz (M)

Comprehensive Pneumology Center (CPC) and Institute of Lung Biology and Disease (ILBD), Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), Munich, Germany.

Travis B Sullivan (TB)

Lahey Hospital & Medical Center, Burlington, MA, USA.

Dawei Sun (D)

Gurdon Institute, University of Cambridge, Cambridge, UK.

Carlos Talavera-López (C)

Cellular Genetics Programme, Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK.

Peng Tan (P)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Jessica Tantivit (J)

Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Center for Cancer Research, Department of Medicine, Massachusetts General Hospital, Boston, MA, USA.
Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital, Charlestown, MA, USA.

Kyle J Travaglini (KJ)

Department of Biochemistry and Wall Center for Pulmonary Vascular Disease, Stanford, CA, USA.

Nathan R Tucker (NR)

Precision Cardiology Laboratory, The Broad Institute, Cambridge, MA, USA.
Masonic Medical Research Institute, Utica, NY, USA.

Katherine A Vernon (KA)

Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA, USA.

Marc H Wadsworth (MH)

Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, USA.
Institute for Medical Engineering and Science, Department of Chemistry & Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA.

Julia Waldman (J)

Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Xiuting Wang (X)

Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, USA.

Ke Xu (K)

Division of Computational Biomedicine, Department of Medicine, Boston University School of Medicine, Boston, MA, USA.

Wenjun Yan (W)

Center for Brain Science, Harvard University, Cambridge, MA, USA.
Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA, USA.

William Zhao (W)

Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, USA.

Carly G K Ziegler (CGK)

Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, USA.
Harvard-MIT Health Sciences and Technology, Institute for Medical Engineering and Science, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA.

Articles similaires

[Redispensing of expensive oral anticancer medicines: a practical application].

Lisanne N van Merendonk, Kübra Akgöl, Bastiaan Nuijen
1.00
Humans Antineoplastic Agents Administration, Oral Drug Costs Counterfeit Drugs

Smoking Cessation and Incident Cardiovascular Disease.

Jun Hwan Cho, Seung Yong Shin, Hoseob Kim et al.
1.00
Humans Male Smoking Cessation Cardiovascular Diseases Female
Humans United States Aged Cross-Sectional Studies Medicare Part C
1.00
Humans Yoga Low Back Pain Female Male

Classifications MeSH