The vitamin D receptor regulates miR-140-5p and targets the MAPK pathway in bone development

Wenting Luo; Lingli Liu; Liping Yang; Yaping Dong; Tianjing Liu; Xiaowei Wei; Dan Liu; Hui Gu; Juan Kong; Zhengwei Yuan; Qun Zhao

doi:10.1016/j.metabol.2018.03.018

The vitamin D receptor regulates miR-140-5p and targets the MAPK pathway in bone development

Metabolism. 2018 Aug:85:139-150. doi: 10.1016/j.metabol.2018.03.018. Epub 2018 Mar 28.

Authors

Wenting Luo¹, Lingli Liu², Liping Yang², Yaping Dong², Tianjing Liu², Xiaowei Wei¹, Dan Liu¹, Hui Gu¹, Juan Kong³, Zhengwei Yuan¹, Qun Zhao⁴

Affiliations

¹ Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital of China Medical University, Shenyang, Liaoning, China.
² Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital of China Medical University, Shenyang, Liaoning, China; Department of Pediatric Orthopedic, Shengjing Hospital of China Medical University, Shenyang, Liaoning, China.
³ Nutrition Department, Shengjing Hospital of China Medical University, Shenyang, Liaoning, China.
⁴ Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital of China Medical University, Shenyang, Liaoning, China; Department of Pediatric Orthopedic, Shengjing Hospital of China Medical University, Shenyang, Liaoning, China. Electronic address: zhaoquncmu@163.com.

PMID: 29604361
DOI: 10.1016/j.metabol.2018.03.018

Abstract

Background: Skeletal development is a complicated process. The status of vitamin D (VD) is closely related to fetal bone development in the embryonic period. Recently, miRNAs have been found to participate in the regulation of skeletal growth and development in several species. However, the mechanisms underlying the interactions among vitamin D, its receptor (VDR), and miRNAs during the process of bone development remain unclear. The aim of this study was to identify miRNAs that are regulated by 1,25(OH)₂D₃ in murine osteoblasts and to analyze the relationship and the effects of VD/VDR and miRNAs in vitro and in vivo.

Methods: We performed miRNA sequencing in murine primary osteoblasts and in an osteoblast cell line treated with 1,25(OH)₂D₃ to identify miRNAs in these cells. After qRT-PCR validation, miR-140-5p was selected for further analysis. We assessed the pathways comprising predicted target genes for several expressed miRNAs, including miR-140-5p, validated predicted target genes in the MAPK pathway by qRT-PCR, and explored the correlation between VD/VDR and miR-140-5p in vitro and in vivo.

Results: 88 miRNAs in murine primary osteoblasts and 49 miRNAs in osteoblast cell line were found to be differentially expressed. MiR-140-5p was upregulated in these 2 types of murine osteoblasts. The expression of miR-140-5p was promoted by 1,25(OH)₂D₃ through transcriptional activation by VDR, with targeted inhibition of MAPK signaling in osteoblasts. A positive correlation between vitamin D/VDR and miR-140-5p was observed in VDR-knockout mice and in 165 human serum specimens. These data show for the first time that VDR transcriptionally activates miR-140-5p. Therefore, the VD/VDR/miR-140-5p/MAPK signaling axis plays an important role in transmitting the effects of 1,25(OH)₂D₃.

Conclusion: Our results demonstrate a novel regulatory mechanism by which miR-140-5p targets the MAPK pathway by means of VD/VDR in vitro and in vivo. These findings provide a new reference for mechanistic research and therapeutic approaches for vitamin D-related bone diseases.

Keywords: 1,25(OH)(2) D(3); Osteoblast; VDR; miR-140-5p; miRNA.

Publication types

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

MeSH terms

Adolescent
Animals
Bone Development / physiology*
Cell Line
Child
Child, Preschool
Female
Humans
Infant
MAP Kinase Signaling System / physiology*
Male
Mice
Mice, Knockout
MicroRNAs / genetics
MicroRNAs / metabolism*
Osteoblasts / metabolism*
Receptors, Calcitriol / genetics
Receptors, Calcitriol / metabolism*
Young Adult

Substances

MIRN140 microRNA, mouse
MicroRNAs
Receptors, Calcitriol