{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,25]],"date-time":"2026-10-25T04:45:43Z","timestamp":1792903543733,"version":"4.5.0"},"reference-count":52,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"21","content-domain":{"domain":["www.ahajournals.org"],"crossmark-restriction":true},"short-container-title":["Circulation"],"published-print":{"date-parts":[[2020,11,24]]},"abstract":"<jats:sec>\n            <jats:title>Background:<\/jats:title>\n            <jats:p>Rupture and erosion of advanced atherosclerotic lesions with a resultant myocardial infarction or stroke are the leading worldwide cause of death. However, we have a limited understanding of the identity, origin, and function of many cells that make up late-stage atherosclerotic lesions, as well as the mechanisms by which they control plaque stability.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Methods:<\/jats:title>\n            <jats:p>We conducted a comprehensive single-cell RNA sequencing of advanced human carotid endarterectomy samples and compared these with single-cell RNA sequencing from murine microdissected advanced atherosclerotic lesions with smooth muscle cell (SMC) and endothelial lineage tracing to survey all plaque cell types and rigorously determine their origin. We further used chromatin immunoprecipitation sequencing (ChIP-seq), bulk RNA sequencing, and an innovative dual lineage tracing mouse to understand the mechanism by which SMC phenotypic transitions affect lesion pathogenesis.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results:<\/jats:title>\n            <jats:p>\n              We provide evidence that SMC-specific Klf4- versus Oct4-knockout showed virtually opposite genomic signatures, and their putative target genes play an important role regulating SMC phenotypic changes. Single-cell RNA sequencing revealed remarkable similarity of transcriptomic clusters between mouse and human lesions and extensive plasticity of SMC- and endothelial cell-derived cells including 7 distinct clusters, most negative for traditional markers. In particular, SMC contributed to a Myh11\n              <jats:sup>-<\/jats:sup>\n              , Lgals3\n              <jats:sup>+<\/jats:sup>\n              population with a chondrocyte-like gene signature that was markedly reduced with SMC-\n              <jats:italic>Klf4<\/jats:italic>\n              knockout. We observed that SMCs that activate Lgals3 compose up to two thirds of all SMC in lesions. However, initial activation of Lgals3 in these cells does not represent conversion to a terminally differentiated state, but rather represents transition of these cells to a unique stem cell marker gene\u2013positive, extracellular matrix-remodeling, \u201cpioneer\u201d cell phenotype that is the first to invest within lesions and subsequently gives rise to at least 3 other SMC phenotypes within advanced lesions, including Klf4-dependent osteogenic phenotypes likely to contribute to plaque calcification and plaque destabilization.\n            <\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions:<\/jats:title>\n            <jats:p>\n              Taken together, these results provide evidence that SMC-derived cells within advanced mouse and human atherosclerotic lesions exhibit far greater phenotypic plasticity than generally believed, with Klf4 regulating transition to multiple phenotypes including Lgals3\n              <jats:sup>+<\/jats:sup>\n              osteogenic cells likely to be detrimental for late-stage atherosclerosis plaque pathogenesis.\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1161\/circulationaha.120.046672","type":"journal-article","created":{"date-parts":[[2020,7,17]],"date-time":"2020-07-17T09:00:12Z","timestamp":1594976412000},"page":"2045-2059","update-policy":"https:\/\/doi.org\/10.1161\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":418,"title":["Stem Cell Pluripotency Genes Klf4 and Oct4 Regulate Complex SMC Phenotypic Changes Critical in Late-Stage Atherosclerotic Lesion Pathogenesis"],"prefix":"10.1161","volume":"142","author":[{"given":"Gabriel F.","family":"Alencar","sequence":"first","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center (G.F.A., K.M.O, S.K., A.N., C.M.W., S.S., C.A.H., R.H., R.A.B., C.A.M., E.R.Z., G.K.O.), University of Virginia, Charlottesville."},{"name":"Department of Biochemistry and Molecular Genetics (G.F.A., K.M.O., C.A.H., R.A.B., S.B.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katherine M.","family":"Owsiany","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center (G.F.A., K.M.O, S.K., A.N., C.M.W., S.S., C.A.H., R.H., R.A.B., C.A.M., E.R.Z., G.K.O.), University of Virginia, Charlottesville."},{"name":"Department of Biochemistry and Molecular Genetics (G.F.A., K.M.O., C.A.H., R.A.B., S.B.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Santosh","family":"Karnewar","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center (G.F.A., K.M.O, S.K., A.N., C.M.W., S.S., C.A.H., R.H., R.A.B., C.A.M., E.R.Z., G.K.O.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katyayani","family":"Sukhavasi","sequence":"additional","affiliation":[{"name":"Department of Cardiac Surgery, Tartu University Hospital, Estonia (K.S.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppe","family":"Mocci","sequence":"additional","affiliation":[{"name":"Integrated Cardio Metabolic Centre, Department of Medicine, Karolinska Institutet, Karolinska Universitetssjukhuset, Huddinge, Sweden (G.M., V.V., J.B.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anh T.","family":"Nguyen","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center (G.F.A., K.M.O, S.K., A.N., C.M.W., S.S., C.A.H., R.H., R.A.B., C.A.M., E.R.Z., G.K.O.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Corey M.","family":"Williams","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center (G.F.A., K.M.O, S.K., A.N., C.M.W., S.S., C.A.H., R.H., R.A.B., C.A.M., E.R.Z., G.K.O.), University of Virginia, Charlottesville."},{"name":"Department of Biomedical Engineering (C.M.W., E.R.Z.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sohel","family":"Shamsuzzaman","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center (G.F.A., K.M.O, S.K., A.N., C.M.W., S.S., C.A.H., R.H., R.A.B., C.A.M., E.R.Z., G.K.O.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5298-4852","authenticated-orcid":true,"given":"Michal","family":"Mokry","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Chemistry and Hematology, Division Laboratories and Pharmacy (M.M., G.P.), University Medical Center Utrecht, University Utrecht, The Netherlands."},{"name":"Department of Cardiology (M.M.), University Medical Center Utrecht, University Utrecht, The Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher A.","family":"Henderson","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center (G.F.A., K.M.O, S.K., A.N., C.M.W., S.S., C.A.H., R.H., R.A.B., C.A.M., E.R.Z., G.K.O.), University of Virginia, Charlottesville."},{"name":"Department of Biochemistry and Molecular Genetics (G.F.A., K.M.O., C.A.H., R.A.B., S.B.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryan","family":"Haskins","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center (G.F.A., K.M.O, S.K., A.N., C.M.W., S.S., C.A.H., R.H., R.A.B., C.A.M., E.R.Z., G.K.O.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard A.","family":"Baylis","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center (G.F.A., K.M.O, S.K., A.N., C.M.W., S.S., C.A.H., R.H., R.A.B., C.A.M., E.R.Z., G.K.O.), University of Virginia, Charlottesville."},{"name":"Department of Biochemistry and Molecular Genetics (G.F.A., K.M.O., C.A.H., R.A.B., S.B.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aloke V.","family":"Finn","sequence":"additional","affiliation":[{"name":"CVPath Institute, Inc, Gaithersburg, MD (A.V.F.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0049-9986","authenticated-orcid":true,"given":"Coleen A.","family":"McNamara","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center (G.F.A., K.M.O, S.K., A.N., C.M.W., S.S., C.A.H., R.H., R.A.B., C.A.M., E.R.Z., G.K.O.), University of Virginia, Charlottesville."},{"name":"School of Medicine, Division of Cardiovascular Medicine, Department of Medicine (C.A.M.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eli R.","family":"Zunder","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center (G.F.A., K.M.O, S.K., A.N., C.M.W., S.S., C.A.H., R.H., R.A.B., C.A.M., E.R.Z., G.K.O.), University of Virginia, Charlottesville."},{"name":"Department of Biomedical Engineering (C.M.W., E.R.Z.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vamsidhar","family":"Venkata","sequence":"additional","affiliation":[{"name":"Integrated Cardio Metabolic Centre, Department of Medicine, Karolinska Institutet, Karolinska Universitetssjukhuset, Huddinge, Sweden (G.M., V.V., J.B.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5345-1022","authenticated-orcid":true,"given":"Gerard","family":"Pasterkamp","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Chemistry and Hematology, Division Laboratories and Pharmacy (M.M., G.P.), University Medical Center Utrecht, University Utrecht, The Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1945-7425","authenticated-orcid":true,"given":"Johan","family":"Bj\u00f6rkegren","sequence":"additional","affiliation":[{"name":"Integrated Cardio Metabolic Centre, Department of Medicine, Karolinska Institutet, Karolinska Universitetssjukhuset, Huddinge, Sweden (G.M., V.V., J.B.)."},{"name":"Department of Genetics and Genomic Sciences (J.B.), Icahn School of Medicine at Mount Sinai, New York."},{"name":"Icahn Institute of Genomics and Multiscale Biology (J.B.), Icahn School of Medicine at Mount Sinai, New York."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3177-4346","authenticated-orcid":true,"given":"Stefan","family":"Bekiranov","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Genetics (G.F.A., K.M.O., C.A.H., R.A.B., S.B.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7119-9657","authenticated-orcid":true,"given":"Gary K.","family":"Owens","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center (G.F.A., K.M.O, S.K., A.N., C.M.W., S.S., C.A.H., R.H., R.A.B., C.A.M., E.R.Z., G.K.O.), University of Virginia, Charlottesville."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"276","reference":[{"key":"e_1_3_5_2_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.114.302721"},{"key":"e_1_3_5_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature10146"},{"key":"e_1_3_5_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/nm.3866"},{"key":"e_1_3_5_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/nm.4109"},{"key":"e_1_3_5_6_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.115.304634"},{"key":"e_1_3_5_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2332"},{"key":"e_1_3_5_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-019-0512-5"},{"key":"e_1_3_5_9_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/bts635"},{"key":"e_1_3_5_10_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btt656"},{"key":"e_1_3_5_11_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-014-0550-8"},{"key":"e_1_3_5_12_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btt703"},{"key":"e_1_3_5_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1923"},{"key":"e_1_3_5_14_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btp352"},{"key":"e_1_3_5_15_2","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2008-9-9-r137"},{"key":"e_1_3_5_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cels.2016.09.002"},{"key":"e_1_3_5_17_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-016-0938-8"},{"key":"e_1_3_5_18_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.4096"},{"key":"e_1_3_5_19_2","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvy084"},{"key":"e_1_3_5_20_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.117.312086"},{"key":"e_1_3_5_21_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.117.312513"},{"key":"e_1_3_5_22_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.117.312509"},{"key":"e_1_3_5_23_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.118.312804"},{"key":"e_1_3_5_24_2","first-page":"261","article-title":"Class II MHC antigen expression in the atherosclerotic plaque: smooth muscle cells express HLA-DR, HLA-DQ and the invariant gamma chain.","volume":"64","author":"Hansson GK","year":"1986","unstructured":"Hansson, GK, Jonasson, L, Holm, J, Claesson-Welsh, L.Class II MHC antigen expression in the atherosclerotic plaque: smooth muscle cells express HLA-DR, HLA-DQ and the invariant gamma chain. Clin Exp Immunol. 1986; 64:261\u2013268","journal-title":"Clin Exp Immunol"},{"key":"e_1_3_5_25_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00418-018-1640-6"},{"key":"e_1_3_5_26_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.110.234914"},{"key":"e_1_3_5_27_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa0912923"},{"key":"e_1_3_5_28_2","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvy010"},{"key":"e_1_3_5_29_2","doi-asserted-by":"publisher","DOI":"10.1002\/dvg.20335"},{"key":"e_1_3_5_30_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-04447-7"},{"key":"e_1_3_5_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2006.07.024"},{"key":"e_1_3_5_32_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.113.005015"},{"key":"e_1_3_5_33_2","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.119.312434"},{"key":"e_1_3_5_34_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-06891-x"},{"key":"e_1_3_5_35_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.180.4.2650"},{"key":"e_1_3_5_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbagen.2005.12.020"},{"key":"e_1_3_5_37_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.neo.2014.09.005"},{"key":"e_1_3_5_38_2","doi-asserted-by":"publisher","DOI":"10.1038\/labinvest.2008.111"},{"key":"e_1_3_5_39_2","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvt206"},{"key":"e_1_3_5_40_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2015.08.008"},{"key":"e_1_3_5_41_2","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.11920"},{"key":"e_1_3_5_42_2","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwy001"},{"key":"e_1_3_5_43_2","doi-asserted-by":"publisher","DOI":"10.1038\/aps.2012.150"},{"key":"e_1_3_5_44_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.426445"},{"key":"e_1_3_5_45_2","doi-asserted-by":"publisher","DOI":"10.1177\/0022034516668321"},{"key":"e_1_3_5_46_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00392-018-1324-1"},{"key":"e_1_3_5_47_2","doi-asserted-by":"publisher","DOI":"10.1007\/s11883-018-0748-4"},{"key":"e_1_3_5_48_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2018.11.014"},{"key":"e_1_3_5_49_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2015.020"},{"key":"e_1_3_5_50_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.22271"},{"key":"e_1_3_5_51_2","doi-asserted-by":"publisher","DOI":"10.1142\/9789813207813_0054"},{"key":"e_1_3_5_52_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1198704"},{"key":"e_1_3_5_53_2","first-page":"03426","article-title":"UMAP: Uniform Manifold Approximation and Projection for Dimension Reduction.","volume":"1802","author":"McInnes L","year":"2018","unstructured":"McInnes, L, Healy, J, Melville, J.UMAP: Uniform Manifold Approximation and Projection for Dimension Reduction. arXiv. 2018; 1802:03426","journal-title":"arXiv"}],"container-title":["Circulation"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.ahajournals.org\/doi\/full\/10.1161\/CIRCULATIONAHA.120.046672","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,13]],"date-time":"2024-05-13T07:37:13Z","timestamp":1715585833000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.ahajournals.org\/doi\/10.1161\/CIRCULATIONAHA.120.046672"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,11,24]]},"references-count":52,"journal-issue":{"issue":"21","published-print":{"date-parts":[[2020,11,24]]}},"alternative-id":["10.1161\/CIRCULATIONAHA.120.046672"],"URL":"https:\/\/doi.org\/10.1161\/circulationaha.120.046672","relation":{},"ISSN":["0009-7322","1524-4539"],"issn-type":[{"value":"0009-7322","type":"print"},{"value":"1524-4539","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,11,24]]},"assertion":[{"value":"2020-03-05","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-06-16","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-07-17","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}