PMID- 27321958 OWN - NLM STAT- MEDLINE DCOM- 20170519 LR - 20171209 IS - 1090-2422 (Electronic) IS - 0014-4827 (Linking) VI - 345 IP - 2 DP - 2016 Jul 15 TI - WNT/beta-catenin signaling promotes VSMCs to osteogenic transdifferentiation and calcification through directly modulating Runx2 gene expression. PG - 206-17 LID - S0014-4827(16)30147-1 [pii] LID - 10.1016/j.yexcr.2016.06.007 [doi] AB - Arterial medial calcification (AMC) is prevalent in patients with chronic kidney disease (CKD) and contributes to elevated risk of cardiovascular events and mortality. Vascular smooth muscle cells (VSMCs) to osteogenic transdifferentiation (VOT) in a high-phosphate environment is involved in the pathogenesis of AMC in CKD. WNT/beta-catenin signaling is indicated to play a crucial role in osteogenesis via promoting Runx2 expression in osteoprogenitor cells, however, its role in Runx2 regulation and VOT remains incompletely clarified. In this study, Runx2 was induced and beta-catenin was activated by high-phosphate in VSMCs. Two forms of active beta-catenin, dephosphorylated on Ser37/Thr41 and phosphorylated on Ser675 sites, were upregulated by high-phosphate. Activation of beta-catenin, through ectopic expression of stabilized beta-catenin, inhibition of GSK-3beta, or WNT-3A protein, induced Runx2 expression, whereas blockade of WNT/beta-catenin signaling with Porcupine (PORCN) inhibitor or Dickkopf-1 (DKK1) protein inhibited Runx2 induction by high-phosphate. WNT-3A promoted osteocalcin expression and calcium deposition in VSMCs, whereas DKK1 ameliorated calcification of VSMCs induced by high-phosphate. Two functional T cell factor (TCF)/lymphoid enhancer-binding factor binding sites were identified in the promoter region of Runx2 gene in VSMCs, which interacted with TCF upon beta-catenin activation. Site-directed mutation of each of them attenuated Runx2 response to beta-catenin, and deletion or destruction of both of them completely abolished this responsiveness. In the aortic tunica media of rats with chronic renal failure, followed by AMC, Runx2 and beta-catenin was induced, and the Runx2 mRNA level was positively associated with the abundance of phosphorylated beta-catenin (Ser675). Collectively, our study suggested that high-phosphate may activate WNT/beta-catenin signaling through different pathways, and the activated WNT/beta-catenin signaling, through direct downstream target Runx2, could play an important role in promoting VOT and AMC. CI - Copyright (c) 2016 The Authors. Published by Elsevier Inc. All rights reserved. FAU - Cai, Ting AU - Cai T AD - Center for Kidney Disease, Second Affiliated Hospital, Nanjing Medical University, Nanjing 210003, China. FAU - Sun, Danqin AU - Sun D AD - Center for Kidney Disease, Second Affiliated Hospital, Nanjing Medical University, Nanjing 210003, China. FAU - Duan, Ying AU - Duan Y AD - Center for Kidney Disease, Second Affiliated Hospital, Nanjing Medical University, Nanjing 210003, China. FAU - Wen, Ping AU - Wen P AD - Center for Kidney Disease, Second Affiliated Hospital, Nanjing Medical University, Nanjing 210003, China. FAU - Dai, Chunsun AU - Dai C AD - Center for Kidney Disease, Second Affiliated Hospital, Nanjing Medical University, Nanjing 210003, China. FAU - Yang, Junwei AU - Yang J AD - Center for Kidney Disease, Second Affiliated Hospital, Nanjing Medical University, Nanjing 210003, China. Electronic address: jwyang@njmu.edu.cn. FAU - He, Weichun AU - He W AD - Center for Kidney Disease, Second Affiliated Hospital, Nanjing Medical University, Nanjing 210003, China. Electronic address: heweichun@njmu.edu.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20160616 PL - United States TA - Exp Cell Res JT - Experimental cell research JID - 0373226 RN - 0 (Core Binding Factor Alpha 1 Subunit) RN - 0 (Glycerophosphates) RN - 0 (TCF Transcription Factors) RN - 0 (beta Catenin) RN - WWH06G87W6 (beta-glycerophosphoric acid) SB - IM MH - Animals MH - Aorta/metabolism MH - Base Sequence MH - Binding Sites MH - *Calcification, Physiologic/drug effects/genetics MH - Cell Transdifferentiation/drug effects/*genetics MH - Core Binding Factor Alpha 1 Subunit/*genetics/metabolism MH - *Gene Expression Regulation/drug effects MH - Glycerophosphates/pharmacology MH - Kidney Failure, Chronic/pathology MH - Muscle, Smooth, Vascular/*cytology MH - Myocytes, Smooth Muscle/drug effects/*metabolism MH - Osteogenesis/drug effects/*genetics MH - Promoter Regions, Genetic/genetics MH - Rats, Sprague-Dawley MH - TCF Transcription Factors/metabolism MH - *Wnt Signaling Pathway/drug effects/genetics MH - beta Catenin/metabolism OTO - NOTNLM OT - Arterial medial calcification OT - High-phosphate OT - Osteogenic transdifferentiation OT - Runx2 OT - Vascular smooth muscle cells OT - WNT/beta-catenin signaling EDAT- 2016/06/21 06:00 MHDA- 2017/05/20 06:00 CRDT- 2016/06/21 06:00 PHST- 2016/01/26 00:00 [received] PHST- 2016/06/10 00:00 [revised] PHST- 2016/06/10 00:00 [accepted] PHST- 2016/06/21 06:00 [entrez] PHST- 2016/06/21 06:00 [pubmed] PHST- 2017/05/20 06:00 [medline] AID - S0014-4827(16)30147-1 [pii] AID - 10.1016/j.yexcr.2016.06.007 [doi] PST - ppublish SO - Exp Cell Res. 2016 Jul 15;345(2):206-17. doi: 10.1016/j.yexcr.2016.06.007. Epub 2016 Jun 16.