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We screen candidate causal CAD genes for regulators of the vascular smooth muscle cell (VSMC) contractile phenotype using a pooled CRISPR\u2013Cas9 knockout library in primary human VSMCs with MYH11 protein levels as the readout. FHL3 (four-and-a-half LIM domains protein 3) emerges as the top repressor of the contractile state. Colocalization analyses in a VSMC biobank and vascular tissues associate a CAD risk allele with reduced FHL3 expression. Biochemical and imaging studies show FHL3 is associated and co-localizes with the transcriptional co-activator MRTFB (myocardin-related transcription factor-B). Functionally, FHL3 overexpression attenuates MRTFB-serum response factor (SRF)-driven induction of contractile markers (MYH11, transgelin), whereas FHL3 knockdown increases their expression in an MRTFB-dependent manner. FHL3 knockdown or MRTFB overexpression reduces VSMC proliferation and migration, and FHL3 overexpression mitigates MRTFB-driven effects on these behaviors. These data identify FHL3 as a regulator of VSMC plasticity that inhibits the contractile phenotype by interfering with MRTFB-SRF signaling, link a CAD-associated genetic signal to the VSMC phenotype, and nominate the FHL3-MRTFB interaction as a potential therapeutic target.<\/jats:p>","DOI":"10.3390\/cells15161475","type":"journal-article","created":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T13:42:17Z","timestamp":1786974137000},"page":"1475","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["FHL3 Regulates Vascular Smooth Muscle Cell Phenotypic Switching Through the MRTFB-SRF Signaling Axis"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-0402-3613","authenticated-orcid":false,"given":"Xiaoxin","family":"Huang","sequence":"first","affiliation":[{"name":"Division of Basic Medicine, Shantou University Medical College, Shantou 515041, China"},{"name":"Cardiovascular-Metabolic Disease Translational Research Programme, Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117599, Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-7451-6610","authenticated-orcid":false,"given":"Heming","family":"Zhang","sequence":"additional","affiliation":[{"name":"Cardiovascular-Metabolic Disease Translational Research Programme, Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117599, Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanhong","family":"Zhang","sequence":"additional","affiliation":[{"name":"Division of Basic Medicine, Shantou University Medical College, Shantou 515041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles U.","family":"Solomon","sequence":"additional","affiliation":[{"name":"Division of Cardiovascular Sciences, University of Leicester, Leicester LE3 9QP, UK"},{"name":"Leicester British Heart Foundation Centre of Research Excellence, University of Leicester, Leicester LE3 9QP, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6402-2546","authenticated-orcid":false,"given":"David G.","family":"McVey","sequence":"additional","affiliation":[{"name":"Division of Cardiovascular Sciences, University of Leicester, Leicester LE3 9QP, UK"},{"name":"Leicester British Heart Foundation Centre of Research Excellence, University of Leicester, Leicester LE3 9QP, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4126-4278","authenticated-orcid":false,"given":"Shu","family":"Ye","sequence":"additional","affiliation":[{"name":"Division of Basic Medicine, Shantou University Medical College, Shantou 515041, China"},{"name":"Cardiovascular-Metabolic Disease Translational Research Programme, Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117599, Singapore"},{"name":"Division of Cardiovascular Sciences, University of Leicester, Leicester LE3 9QP, UK"},{"name":"The First Affiliated Hospital of Shantou University Medical College, Shantou 515041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2361","DOI":"10.1016\/j.jacc.2022.11.005","article-title":"The Global Burden of Cardiovascular Diseases and Risk: A Compass for Future Health","volume":"80","author":"Vaduganathan","year":"2022","journal-title":"J. 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