Circulation Research RES Serine Protease Activation Essential for Endothelial-Mesenchymal Transition in Vascular Calcification Serine Proteases in Vascular Calcification CIRCRES/2015/306751 CIRCRES/2015/306751 10.1161/CIRCRESAHA.115.306751 117 10/09/15 9 Bridges-Lyman Gemma 410-3275005 410-3279322 Griendling, Kathy Emory University Dr. Yucheng Yao yyao@mednet.ucla.edu Dr. University of California, Los Angeles 650 Charles E.Young Dr. So CHS A2-237 Los Angeles California 90095 UNITED STATES 310-2674951 310-2068553 11806 Jiayi Yao University of California, Los Angeles JYao@mednet.ucla.edu 217598 Pierre Guihard University of California, Los Angeles PGuihard@mednet.ucla.edu 220405 Ana M Blazquez-Medela University of California, Los Angeles ABlazquezMedela@mednet.ucla.edu 217597 Yina Guo University of California, Los Angeles yinaguo2012@gmail.com 276807 Jeremiah H Moon University of California, Los Angeles jeremymoon93@gmail.com 290778 Medet Jumabay University of California, Los Angeles mjumabay@mednet.ucla.edu 113583 Kristina I Bostrom University of California, Los Angeles kbostrom@mednet.ucla.edu 127547 Yucheng Yao University of California, Los Angeles yyao@mednet.ucla.edu 11806 04/24/2015 04/24/2015 08/05/2015 08/11/2015 08/12/2015 Regular Article Vascular Biology Matrix Gla protein serine protease <p><i><b><u>Rationale:</u></b></i> Endothelial cells have the ability to undergo endothelial-mesenchymal transitions (EndMTs), by which they acquire a mesenchymal phenotype and stem-cell like characteristics. We previously found that EndMTs ocurred in the endothelium deficient in matrix Gla protein (MGP) enabling endothelial cells to contribute cells to vascular calcification. However, the mechanism responsible for initiating EndMTs is not fully understood. </p><p><i><b><u>Objective:</u></b></i> To determine the role of specific serine proteases and sex determining region Y-box 2 (Sox2) in the initiation of EndMTs. </p><p><i><b><u>Methods and Results:</u></b></i> In this study, we used in vivo and in vitro models of vascular calcification to demonstrate that serine proteases and Sox2 are essential for the initiation of EndMTs in MGP-deficient endothelium. We showed that expression of a group of specific serine proteases was highly induced in endothelial cells at sites of vascular calcification in Mgp null aortas. Treatment with serine protease inhibitors decreased both stem-cell marker expression and vascular calcification. In human aortic endothelial cells, this group of serine proteases also induced EndMTs, and the activation of proteases was mediated by Sox2. Knockdown of the serine proteases or Sox2 diminished EndMTs and calcification. Endothelial-specific deletion of Sox2 decreased expression of stem-cell markers and aortic calcification in MGP-deficient mice. </p><p><i><b><u>Conclusions:</u></b></i> Our results suggest that Sox2-mediated activation of specific serine proteases is essential for initiating EndMTs, and thus, may provide new therapeutic targets for treating vascular calcification. </p> 0 0 1 7 8 no yes CircRes_CIRCRES-2015-306751.xml CircRes_CIRCRES-2015-306751_file1.doc CircRes_CIRCRES-2015-306751_supp1.pdf

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CircRes_CIRCRES-2015-306751_merge.pdf CircRes_CIRCRES-2015-306751_Guo_276807_disclosure.pdf CircRes_CIRCRES-2015-306751_Guo_276807_copyright.pdf CircRes_CIRCRES-2015-306751_Yao_11806_disclosure.pdf CircRes_CIRCRES-2015-306751_Yao_11806_copyright.pdf CircRes_CIRCRES-2015-306751_Jumabay_113583_disclosure.pdf CircRes_CIRCRES-2015-306751_Jumabay_113583_copyright.pdf CircRes_CIRCRES-2015-306751_Guihard_220405_disclosure.pdf CircRes_CIRCRES-2015-306751_Guihard_220405_copyright.pdf CircRes_CIRCRES-2015-306751_Yao_217598_disclosure.pdf CircRes_CIRCRES-2015-306751_Yao_217598_copyright.pdf CircRes_CIRCRES-2015-306751_Moon_290778_disclosure.pdf CircRes_CIRCRES-2015-306751_Moon_290778_copyright.pdf CircRes_CIRCRES-2015-306751_Blazquez-Medela_217597_disclosure.pdf CircRes_CIRCRES-2015-306751_Blazquez-Medela_217597_copyright.pdf CircRes_CIRCRES-2015-306751_Bostrom_127547_disclosure.pdf CircRes_CIRCRES-2015-306751_Bostrom_127547_copyright.pdf PAP: 08/11/15 Please note that the authors have agreed to pay $425 in excess page charges. Funding: Howard Hughes Medical Institute (HHMI): No National Institutes of Health (NIH): Yes Not applicable for this manuscript: No Other: Yes Wellcome Trust: No Subject Codes: [145] Genetically altered mice [147] Growth factors/cytokines [95] Endothelium/vascular type/nitric oxide [97] Other vascular biology gbridgeslyman