• Please log in or register to access this feature.

Original Article

Vitamin D activation of functionally distinct regulatory miRNAs in primary human osteoblasts

  1. Thomas S Lisse1,†,
  2. Rene F Chun1,
  3. Sandra Rieger3,
  4. John S Adams1,2,
  5. Martin Hewison1,2,*

Article first published online: 21 MAY 2013

DOI: 10.1002/jbmr.1882

Issue

Journal of Bone and Mineral Research

Journal of Bone and Mineral Research

Volume 28, Issue 6, pages 1478–1488, June 2013

Additional Information

How to Cite

Lisse, T. S., Chun, R. F., Rieger, S., Adams, J. S. and Hewison, M. (2013), Vitamin D activation of functionally distinct regulatory miRNAs in primary human osteoblasts. J Bone Miner Res, 28: 1478–1488. doi: 10.1002/jbmr.1882

Author Information

  1. 1

    Orthopaedic Hospital Research Center, University of California Los Angeles, Los Angeles, CA, USA

  2. 2

    Molecular Biology Institute, University of California Los Angeles, Los Angeles, CA, USA

  3. 3

    Mount Desert Island Biological Laboratory, Salisbury Cove, ME, USA

  1. †

    TSL, Massachusetts General Hospital, Endocrine Unit, Harvard School of Medicine, Boston, MA 02114, USA

*Address correspondence to: Martin Hewison, PhD, Orthopaedic Hospital Research Center, University of California Los Angeles, 615 Charles E. Young Drive South, Los Angeles, CA 90095, USA; E-mail: mhewison@mednet.ucla.edu.

Publication History

  1. Issue published online: 21 MAY 2013
  2. Article first published online: 21 MAY 2013
  3. Accepted manuscript online: 29 JAN 2013 01:53PM EST
  4. Manuscript Accepted: 17 JAN 2013
  5. Manuscript Revised: 3 JAN 2013
  6. Manuscript Received: 16 MAY 2012

Funded by

  • NIH. Grant Number: 2R01AR037399-21

SEARCH

SEARCH BY CITATION

ARTICLE TOOLS

View Full Article with Supporting Information (HTML) Get PDF (1873K)

Keywords:

  • VITAMIN D;
  • OSTEOBLAST;
  • MICRORNA;
  • TRANSCRIPTION;
  • TRANSLATION;
  • POST-TRANSCRIPTIONAL

ABSTRACT

When bound to the vitamin D receptor (VDR), the active form of vitamin D, 1,25-dihydroxyvitamin D (1,25D) is a potent regulator of osteoblast transcription. Less clear is the impact of 1,25D on posttranscriptional events in osteoblasts, such as the generation and action of microRNAs (miRNAs). Microarray analysis using replicate (n = 3) primary cultures of human osteoblasts (HOBs) identified human miRNAs that were differentially regulated by >1.5-fold following treatment with 1,25D (10 nM, 6 hours), which included miRNAs 637 and 1228. Quantitative reverse transcription PCR analyses showed that the host gene for miR-1228, low-density lipoprotein receptor-related protein 1 (LRP1), was coinduced with miR-1228 in a dose-dependent fashion following treatment with 1,25D (0.1–10 nM, 6 hours). By contrast, the endogenous host gene for miR-637, death-associated protein kinase 3 (DAPK3), was transcriptionally repressed by following treatment with 1,25D. Analysis of two potential targets for miR-637 and miR-1228 in HOB, type IV collagen (COL4A1) and bone morphogenic protein 2 kinase (BMP2K), respectively, showed that 1,25D-mediates suppression of these targets via distinct mechanisms. In the case of miR-637, suppression of COL4A1 appears to occur via decreased levels of COL4A1 mRNA. By contrast, suppression of BMP2K by miR-1228 appears to occur by inhibition of protein translation. In mature HOBs, small interfering RNA (siRNA) inactivation of miR-1228 alone was sufficient to abrogate 1,25D-mediated downregulation of BMP2K protein expression. This was associated with suppression of prodifferentiation responses to 1,25D in HOB, as represented by parallel decrease in osteocalcin and alkaline phosphatase expression. These data show for the first time that the effects of 1,25D on human bone cells are not restricted to classical VDR-mediated transcriptional responses but also involve miRNA-directed posttranscriptional mechanisms.

View Full Article with Supporting Information (HTML) Get PDF (1873K)