WISP-1/CCN4 regulates osteogenesis by enhancing BMP-2 activity

Mitsuaki Ono, Colette A. Inkson, Tina M. Kilts, Marian F. Young

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

Wnt-induced secreted protein 1 (WISP-1/CCN4) is a member of the CCN family that is highly expressed in skeletal tissue and in osteoprogenitor cells induced to differentiate in vitro. To determine the function of WISP-1 during osteogeneis, osteogenic bone marrow stromal cells (BMSCs) were transduced with WISP-1 adenovirus (adWISP-1) in the presence or absence of bone morphogenetic protein 2 (BMP-2) adenovirus (adBMP-2). WISP-1 overexpression enhanced the ability of BMP-2 to direct BMSCs toward osteogenic differentiation and appeared to work by stimulating Smad-1/5/8 phosphorylation and activation. The ability of WISP-1 to enhance BMP-2 activity also was shown in vivo using an ectopic osteogenesis assay with BMSCs transduced with WISP-1, BMP-2, or both. When BMSCs were infected with lentivirus containing human WISP1 shRNA, they formed less bone in vivo and were less responsive to BMP-2, confirming that WISP-1 and BMP-2 have a functional interaction. Immunoprecipitation (IP) and Western blot analysis showed that WISP-1 bound directly to BMP-2 and showed that WISP-1 increased BMP-2 binding to hBMSCs in a dose-dependent fashion. To understand how WISP-1 enhanced BMP-2 signaling, the influence of WISP-1 on integrin expression was analyzed. WISP-1 induced the mRNA and protein levels of α5-integrin and, further, was found to bind to it. Antibody-blocking experiments showed that the BMP-2 binding to BMSCs that was enhanced by WISP-1 was completely neutralized by treatment with anti-integrin α5β1 antibody. Pilot studies and the use of transgenic mice that overexpressed human WISP-1 in preosteoblasts had increased bone mineral density (BMD), trabecular thickness, and bone volume (BV/TV) over wild-type controls, supporting observations using human osteoprogenitors that WISP-1 has a positive influence on osteogenesis in vivo. In conclusion, these studies show, for the first time, that WISP-1 has a positive influence on bone cell differentiation and function and may work by enhancing the effects of BMP-2 to increase osteogenesis through a mechanism potentially involving binding to integrin α5β1.

Original languageEnglish
Pages (from-to)193-208
Number of pages16
JournalJournal of Bone and Mineral Research
Volume26
Issue number1
DOIs
Publication statusPublished - Jan 2011
Externally publishedYes

Fingerprint

Bone Morphogenetic Protein 2
Osteogenesis
Mesenchymal Stromal Cells
Integrins
Aptitude
Protein Binding
Adenoviridae
Bone and Bones
Lentivirus
Blocking Antibodies
Immunoprecipitation
Bone Density
Small Interfering RNA
Transgenic Mice
Cell Differentiation
Proteins
Western Blotting
Phosphorylation

Keywords

  • BMP
  • CCN
  • CYTOKINES
  • GROWTH FACTORS
  • OSTEOPROGENITORS
  • WISP1

ASJC Scopus subject areas

  • Orthopedics and Sports Medicine
  • Endocrinology, Diabetes and Metabolism

Cite this

WISP-1/CCN4 regulates osteogenesis by enhancing BMP-2 activity. / Ono, Mitsuaki; Inkson, Colette A.; Kilts, Tina M.; Young, Marian F.

In: Journal of Bone and Mineral Research, Vol. 26, No. 1, 01.2011, p. 193-208.

Research output: Contribution to journalArticle

Ono, Mitsuaki ; Inkson, Colette A. ; Kilts, Tina M. ; Young, Marian F. / WISP-1/CCN4 regulates osteogenesis by enhancing BMP-2 activity. In: Journal of Bone and Mineral Research. 2011 ; Vol. 26, No. 1. pp. 193-208.
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