{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T17:49:38Z","timestamp":1772560178006,"version":"3.50.1"},"reference-count":70,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2021,5,21]],"date-time":"2021-05-21T00:00:00Z","timestamp":1621555200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"European Union\u2019s Horizon 2020\/Marie Sklodowska-Curie program INTRICARE","award":["722609"],"award-info":[{"award-number":["722609"]}]},{"name":"Swedish Heart and Lung Foundation","award":["20180036, 20170584, 20180244, 201602877, 20180247"],"award-info":[{"award-number":["20180036, 20170584, 20180244, 201602877, 20180247"]}]},{"DOI":"10.13039\/501100004359","name":"Swedish Research Council","doi-asserted-by":"publisher","award":["2017-01070, 2019-02027"],"award-info":[{"award-number":["2017-01070, 2019-02027"]}],"id":[{"id":"10.13039\/501100004359","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Dutch Thrombosis Society","award":["2014.02"],"award-info":[{"award-number":["2014.02"]}]},{"name":"NWO ZonMw","award":["MKMD 40-42600-98-13007"],"award-info":[{"award-number":["MKMD 40-42600-98-13007"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Calcification is a prominent feature of late-stage atherosclerosis, but the mechanisms driving this process are unclear. Using a biobank of carotid endarterectomies, we recently showed that Proteoglycan 4 (PRG4) is a key molecular signature of calcified plaques, expressed in smooth muscle cell (SMC) rich regions. Here, we aimed to unravel the PRG4 role in vascular remodeling and intimal calcification. PRG4 expression in human carotid endarterectomies correlated with calcification assessed by preoperative computed tomographies. PRG4 localized to SMCs in early intimal thickening, while in advanced lesions it was found in the extracellular matrix, surrounding macro-calcifications. In experimental models, Prg4 was upregulated in SMCs from partially ligated ApoE\u2212\/\u2212 mice and rat carotid intimal hyperplasia, correlating with osteogenic markers and TGFb1. Furthermore, PRG4 was enriched in cells positive for chondrogenic marker SOX9 and around plaque calcifications in ApoE\u2212\/\u2212 mice on warfarin. In vitro, PRG4 was induced in SMCs by IFNg, TGFb1 and calcifying medium, while SMC markers were repressed under calcifying conditions. Silencing experiments showed that PRG4 expression was driven by transcription factors SMAD3 and SOX9. Functionally, the addition of recombinant human PRG4 increased ectopic SMC calcification, while arresting cell migration and proliferation. Mechanistically, it suppressed endogenous PRG4, SMAD3 and SOX9, and restored SMC markers\u2019 expression. PRG4 modulates SMC function and osteogenic phenotype during intimal remodeling and macro-calcification in response to TGFb1 signaling, SMAD3 and SOX9 activation. The effects of PRG4 on SMC phenotype and calcification suggest its role in atherosclerotic plaque stability, warranting further investigations.<\/jats:p>","DOI":"10.3390\/cells10061276","type":"journal-article","created":{"date-parts":[[2021,5,21]],"date-time":"2021-05-21T13:15:15Z","timestamp":1621602915000},"page":"1276","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Proteoglycan 4 Modulates Osteogenic Smooth Muscle Cell Differentiation during Vascular Remodeling and Intimal Calcification"],"prefix":"10.3390","volume":"10","author":[{"given":"Till","family":"Seime","sequence":"first","affiliation":[{"name":"Vascular Surgery, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17164 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Asim Cengiz","family":"Akbulut","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, CARIM, Maastricht University, 6229 ER Maastricht, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Moritz Lindquist","family":"Liljeqvist","sequence":"additional","affiliation":[{"name":"Vascular Surgery, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17164 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antti","family":"Siika","sequence":"additional","affiliation":[{"name":"Vascular Surgery, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17164 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8878-6436","authenticated-orcid":false,"given":"Hong","family":"Jin","sequence":"additional","affiliation":[{"name":"Vascular Surgery, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17164 Stockholm, Sweden"},{"name":"Department of Medicine, Karolinska Institutet, 17164 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1832-5423","authenticated-orcid":false,"given":"Greg","family":"Winski","sequence":"additional","affiliation":[{"name":"Department of Medicine, Karolinska Institutet, 17164 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rick H.","family":"van Gorp","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, CARIM, Maastricht University, 6229 ER Maastricht, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eva","family":"Karl\u00f6f","sequence":"additional","affiliation":[{"name":"Vascular Surgery, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17164 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mariette","family":"Lengquist","sequence":"additional","affiliation":[{"name":"Vascular Surgery, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17164 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0786-4835","authenticated-orcid":false,"given":"Andrew J.","family":"Buckler","sequence":"additional","affiliation":[{"name":"Vascular Surgery, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17164 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Malin","family":"Kronqvist","sequence":"additional","affiliation":[{"name":"Vascular Surgery, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17164 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Olivia J.","family":"Waring","sequence":"additional","affiliation":[{"name":"Department of Pathology, CARIM, Maastricht University Medical Center, 6200 MD Maastricht, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jan H. N.","family":"Lindeman","sequence":"additional","affiliation":[{"name":"Department of Surgery, Leiden University Medical Center, 2300 RC Leiden, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Erik A. L.","family":"Biessen","sequence":"additional","affiliation":[{"name":"Department of Pathology, CARIM, Maastricht University Medical Center, 6200 MD Maastricht, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5228-2634","authenticated-orcid":false,"given":"Lars","family":"Maegdefessel","sequence":"additional","affiliation":[{"name":"Department of Medicine, Karolinska Institutet, 17164 Stockholm, Sweden"},{"name":"Department for Vascular and Endovascular Surgery, Klinikum rechts der Isar, Technische Universit\u00e4t M\u00fcnchen, 81679 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anton","family":"Razuvaev","sequence":"additional","affiliation":[{"name":"Vascular Surgery, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17164 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7867-6957","authenticated-orcid":false,"given":"Leon J.","family":"Schurgers","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, CARIM, Maastricht University, 6229 ER Maastricht, The Netherlands"},{"name":"Institute of Experimental Medicine and Systems Biology, RWTH Aachen University, 52062 Aachen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ulf","family":"Hedin","sequence":"additional","affiliation":[{"name":"Vascular Surgery, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17164 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ljubica","family":"Matic","sequence":"additional","affiliation":[{"name":"Vascular Surgery, Department of Molecular Medicine and Surgery, Karolinska Institutet, 17164 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,5,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1282","DOI":"10.1161\/ATVBAHA.108.179739","article-title":"Concept of vulnerable\/unstable plaque","volume":"30","author":"Finn","year":"2010","journal-title":"Arterioscler. 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