Functional cooperation between vitamin D receptor and Runx2 in vitamin D-induced vascular calcification

Min-Su Han; Xiangguo Che; Gyoung-ho Cho; Hye-Ri Park; Kyung-Eun Lim; Na-Rae Park; Jung-Sook Jin; Youn-Kwan Jung; Jae-Hwan Jeong; In-Kyu Lee; Shigeaki Kato; Je-Yong Choi

doi:10.1371/journal.pone.0083584

Functional cooperation between vitamin D receptor and Runx2 in vitamin D-induced vascular calcification

PLoS One. 2013 Dec 12;8(12):e83584. doi: 10.1371/journal.pone.0083584. eCollection 2013.

Authors

Affiliations

¹ Department of Biochemistry and Cell Biology, WCU and BK21 plus programs, CMRC, Skeletal Diseases Genome Research Center, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
² Department of Internal Medicine, Division of Endocrinology, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
³ Institute of Molecular and Cellular Biosciences, University of Tokyo, Tokyo, Japan.

Abstract

The transdifferentiation of vascular smooth muscle cells (VSMCs) into osteoblast-like cells has been implicated in the context of vascular calcification. We investigated the roles of vitamin D receptor (Vdr) and runt-related transcription factor 2 (Runx2) in the osteoblastic differentiation of VSMCs in response to vitamin D3 using in vitro VSMCs cultures and in vivo in Vdr knockout (Vdr(-/-)) and Runx2 carboxy-terminus truncated heterozygous (Runx2(+/ΔC)) mice. Treatment of VSMCs with active vitamin D3 promoted matrix mineral deposition, and increased the expressions of Vdr, Runx2, and of osteoblastic genes but decreased the expression of smooth muscle myosin heavy chain in primary VSMCs cultures. Immunoprecipitation experiments suggested an interaction between Vdr and Runx2. Furthermore, silencing Vdr or Runx2 attenuated the procalcific effects of vitamin D3. Functional cooperation between Vdr and Runx2 in vascular calcification was also confirmed in in vivo mouse models. Vascular calcification induced by high-dose vitamin D3 was completely inhibited in Vdr(-/-) or Runx2(+/ΔC) mice, despite elevated levels of serum calcium or alkaline phosphatase. Collectively, these findings suggest that functional cooperation between Vdr and Runx2 is necessary for vascular calcification in response to vitamin D3.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Bone Density Conservation Agents / adverse effects*
Bone Density Conservation Agents / pharmacology
Cells, Cultured
Cholecalciferol / adverse effects*
Cholecalciferol / pharmacology
Core Binding Factor Alpha 1 Subunit / genetics
Core Binding Factor Alpha 1 Subunit / metabolism*
Mice
Mice, Knockout
Muscle, Smooth, Vascular / metabolism*
Muscle, Smooth, Vascular / pathology
Myocytes, Smooth Muscle / metabolism*
Myocytes, Smooth Muscle / pathology
Rats
Receptors, Calcitriol / genetics
Receptors, Calcitriol / metabolism*
Vascular Calcification* / chemically induced
Vascular Calcification* / genetics
Vascular Calcification* / metabolism
Vascular Calcification* / pathology

Substances

Bone Density Conservation Agents
Core Binding Factor Alpha 1 Subunit
Receptors, Calcitriol
Runx2 protein, mouse
Runx2 protein, rat
Cholecalciferol

Grants and funding

This research was supported by a grant of the Korean Health Technology R&D Project, Ministry of Health & Welfare, Republic of Korea (A111487), the WCU (World Class University) program (Grant no. R32-10064), and Basic Science Research Program (Grant no. 2012R1A6A3A01019166) through the National Research Foundation of Korea funded by the Korean Ministry of Education, Science and Technology. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.