NG2/CSPG4, CD146/MCAM and VAP1/AOC3 are regulated by myocardin-related transcription factors in smooth muscle cells

• Published in: Scientific reports Vol. 11; no. 1; pp. 5955 - 17
• Main Authors: Rippe, Catarina, Morén, Björn, Liu, Li, Stenkula, Karin G., Mustaniemi, Johan, Wennström, Malin, Swärd, Karl
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 16.03.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 631/114; 631/337; 631/443; 631/45; 631/80; 692/53; Actin; Amine Oxidase (Copper-Containing) - genetics; Basic Medicine; Binding sites; Bioinformatics; CD146 Antigen - genetics; Cell Adhesion Molecules - genetics; Cell and Molecular Biology; Cell Differentiation; Cell Line; Cell- och molekylärbiologi; Chondroitin Sulfate Proteoglycans - genetics; Chromatin remodeling; Endothelial cells; Gene Expression Regulation; Gene Knockdown Techniques; Humanities and Social Sciences; Humans; Immunohistochemistry; Medical and Health Sciences; Medicin och hälsovetenskap; Medicinska och farmaceutiska grundvetenskaper; Membrane Proteins - genetics; multidisciplinary; Muscle, Smooth, Vascular - cytology; Muscle, Smooth, Vascular - metabolism; Myocytes, Smooth Muscle - cytology; Myocytes, Smooth Muscle - metabolism; Neuroimaging; Nuclear Proteins - metabolism; Organ Specificity; Pericytes; Protein Binding; Science; Science (multidisciplinary); Serum response factor; Smooth muscle; Ternary complex factors; Trans-Activators - metabolism; Transcription activation; Transcription factors; Transcription Factors - genetics; Transcription Factors - metabolism
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-021-85335-x

The present work addressed the hypothesis that NG2/ CSPG4 , CD146/ MCAM , and VAP1/ AOC3 are target genes of myocardin-related transcription factors (MRTFs: myocardin/ MYOCD , MRTF-A/ MKL1 , MRTF-B/ MKL2 ) and serum response factor ( SRF ). Using a bioinformatics approach, we found that CSPG4 , MCAM , and AOC3 correlate with MYOCD , MRTF-A/ MKL1 , and SRF across human tissues. No other transcription factor correlated as strongly with these transcripts as SRF . Overexpression of MRTFs increased both mRNA and protein levels of CSPG4 , MCAM , and AOC3 in cultured human smooth muscle cells (SMCs). Imaging confirmed increased staining for CSPG4, MCAM, and AOC3 in MRTF-A/ MKL1 -transduced cells. MRTFs exert their effects through SRF, and the MCAM and AOC3 gene loci contained binding sites for SRF. SRF silencing reduced the transcript levels of these genes, and time-courses of induction paralleled the direct target ACTA2 . MRTF-A/ MKL1 increased the activity of promoter reporters for MCAM and AOC3 , and transcriptional activation further depended on the chromatin remodeling enzyme KDM3A. CSPG4 , MCAM , and AOC3 responded to the MRTF-SRF inhibitor CCG-1423, to actin dynamics, and to ternary complex factors. Coincidental detection of these proteins should reflect MRTF-SRF activity, and beyond SMCs, we observed co-expression of CD146/ MCAM , NG2/ CSPG4 , and VAP1/ AOC3 in pericytes and endothelial cells in the human brain. This work identifies highly responsive vascular target genes of MRTF-SRF signaling that are regulated via a mechanism involving KDM3A.