α9β1: A novel osteoclast integrin that regulates osteoclast formation and function

Hongwei Rao, Ganwei Lu, Hiroshi Kajiya, Veronica Garcia-Palacios, Noriyoshi Kurihara, Judy Anderson, Ken Patrene, Dean Sheppard, Harry C. Blair, Jolene J. Windle, Sun Jin Choi, G. David Roodman

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

We identified a previously unknown integrin, α9β 1, on OCLs and their precursors. Antibody to α9 inhibited OCL formation in human marrow cultures, and OCLs from α9 knockout mice had a defect in actin ring reorganization and an impaired bone resorption capacity. Introduction: Integrins play important roles in osteoclast (OCL) formation and function. Mature OCLs mainly express αvβ3 integrin, a heterodimer adhesion receptor that has been implicated in osteoclastic bone resorption. We identified ADAMS, a disintegrin and metalloproteinase, as a novel stimulator of OCL differentiation and showed that the disintegrin domain of ADAMS mediated its effects on OCL formation. Because the disintegrin domain of ADAMS does not bind Arg-Gly-Asp (RGD) sequences, we determined which integrin bound ADAMS and characterized its role in OCL formation and activity. Materials and Methods: Chinese hamster ovary cells (CHO) expressing different integrin subunits were tested for their capacity to bind the disintegrin domain of ADAMS. Mouse or human bone marrow cells and purified OCL precursors were tested for αvβ 1 integrin expression by Western blot, immunocytochemistry, and real-time RT-PCR. A monoclonal antibody to human α9 was used to block α9β1 on OCL precursors stimulated by 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D 3] or RANKL. Vertebrae of 7-day-old α9-/- mice and wildtype (WT) littermates were compared using bone histomorphometry and 3D μCT analysis. Results: α9 integrin was expressed by mouse and human bone marrow-derived OCLs and their precursors. Importantly, the anti-α9 antibody inhibited human OCL formation stimulated by 1α,25(OH)2D3 or RANKL dose-dependently. Furthermore, analysis of OCLs formed in marrow cultures from α9 -/- mice showed that the OCLs formed were more contracted and formed significantly less bone resorption pits on dentin slices. Histologic analysis of α9-/- vertebrae showed thickened trabecular regions and retained cartilage within vertebral bodies of α9 -/- mice. 3D μCT analysis of α9-/- vertebrae also showed a significant increase in trabecular bone volume/total tissue volume and a tendency for decreased trabecular separation compared with WT mice. Conclusions: These results support a previously unknown role for α9β1 integrin in OCL formation and function.

Original languageEnglish (US)
Pages (from-to)1657-1665
Number of pages9
JournalJournal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
Volume21
Issue number10
DOIs
StatePublished - Oct 2006
Externally publishedYes

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Osteoclasts
Integrins
Disintegrins
Bone Resorption
Spine
Bone Marrow
Calcitriol
Metalloproteases
Dentin
Cricetulus
Knockout Mice
Bone Marrow Cells
Cartilage
Actins
Real-Time Polymerase Chain Reaction
Anti-Idiotypic Antibodies
Ovary
Western Blotting
Immunohistochemistry
Monoclonal Antibodies

Keywords

  • α knockout mice
  • αβ integrin
  • ADAM8
  • Osteoclast

ASJC Scopus subject areas

  • Surgery

Cite this

α9β1 : A novel osteoclast integrin that regulates osteoclast formation and function. / Rao, Hongwei; Lu, Ganwei; Kajiya, Hiroshi; Garcia-Palacios, Veronica; Kurihara, Noriyoshi; Anderson, Judy; Patrene, Ken; Sheppard, Dean; Blair, Harry C.; Windle, Jolene J.; Choi, Sun Jin; Roodman, G. David.

In: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, Vol. 21, No. 10, 10.2006, p. 1657-1665.

Research output: Contribution to journalArticle

Rao, Hongwei ; Lu, Ganwei ; Kajiya, Hiroshi ; Garcia-Palacios, Veronica ; Kurihara, Noriyoshi ; Anderson, Judy ; Patrene, Ken ; Sheppard, Dean ; Blair, Harry C. ; Windle, Jolene J. ; Choi, Sun Jin ; Roodman, G. David. / α9β1 : A novel osteoclast integrin that regulates osteoclast formation and function. In: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research. 2006 ; Vol. 21, No. 10. pp. 1657-1665.
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abstract = "We identified a previously unknown integrin, α9β 1, on OCLs and their precursors. Antibody to α9 inhibited OCL formation in human marrow cultures, and OCLs from α9 knockout mice had a defect in actin ring reorganization and an impaired bone resorption capacity. Introduction: Integrins play important roles in osteoclast (OCL) formation and function. Mature OCLs mainly express αvβ3 integrin, a heterodimer adhesion receptor that has been implicated in osteoclastic bone resorption. We identified ADAMS, a disintegrin and metalloproteinase, as a novel stimulator of OCL differentiation and showed that the disintegrin domain of ADAMS mediated its effects on OCL formation. Because the disintegrin domain of ADAMS does not bind Arg-Gly-Asp (RGD) sequences, we determined which integrin bound ADAMS and characterized its role in OCL formation and activity. Materials and Methods: Chinese hamster ovary cells (CHO) expressing different integrin subunits were tested for their capacity to bind the disintegrin domain of ADAMS. Mouse or human bone marrow cells and purified OCL precursors were tested for αvβ 1 integrin expression by Western blot, immunocytochemistry, and real-time RT-PCR. A monoclonal antibody to human α9 was used to block α9β1 on OCL precursors stimulated by 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D 3] or RANKL. Vertebrae of 7-day-old α9-/- mice and wildtype (WT) littermates were compared using bone histomorphometry and 3D μCT analysis. Results: α9 integrin was expressed by mouse and human bone marrow-derived OCLs and their precursors. Importantly, the anti-α9 antibody inhibited human OCL formation stimulated by 1α,25(OH)2D3 or RANKL dose-dependently. Furthermore, analysis of OCLs formed in marrow cultures from α9 -/- mice showed that the OCLs formed were more contracted and formed significantly less bone resorption pits on dentin slices. Histologic analysis of α9-/- vertebrae showed thickened trabecular regions and retained cartilage within vertebral bodies of α9 -/- mice. 3D μCT analysis of α9-/- vertebrae also showed a significant increase in trabecular bone volume/total tissue volume and a tendency for decreased trabecular separation compared with WT mice. Conclusions: These results support a previously unknown role for α9β1 integrin in OCL formation and function.",
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author = "Hongwei Rao and Ganwei Lu and Hiroshi Kajiya and Veronica Garcia-Palacios and Noriyoshi Kurihara and Judy Anderson and Ken Patrene and Dean Sheppard and Blair, {Harry C.} and Windle, {Jolene J.} and Choi, {Sun Jin} and Roodman, {G. David}",
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TY - JOUR

T1 - α9β1

T2 - A novel osteoclast integrin that regulates osteoclast formation and function

AU - Rao, Hongwei

AU - Lu, Ganwei

AU - Kajiya, Hiroshi

AU - Garcia-Palacios, Veronica

AU - Kurihara, Noriyoshi

AU - Anderson, Judy

AU - Patrene, Ken

AU - Sheppard, Dean

AU - Blair, Harry C.

AU - Windle, Jolene J.

AU - Choi, Sun Jin

AU - Roodman, G. David

PY - 2006/10

Y1 - 2006/10

N2 - We identified a previously unknown integrin, α9β 1, on OCLs and their precursors. Antibody to α9 inhibited OCL formation in human marrow cultures, and OCLs from α9 knockout mice had a defect in actin ring reorganization and an impaired bone resorption capacity. Introduction: Integrins play important roles in osteoclast (OCL) formation and function. Mature OCLs mainly express αvβ3 integrin, a heterodimer adhesion receptor that has been implicated in osteoclastic bone resorption. We identified ADAMS, a disintegrin and metalloproteinase, as a novel stimulator of OCL differentiation and showed that the disintegrin domain of ADAMS mediated its effects on OCL formation. Because the disintegrin domain of ADAMS does not bind Arg-Gly-Asp (RGD) sequences, we determined which integrin bound ADAMS and characterized its role in OCL formation and activity. Materials and Methods: Chinese hamster ovary cells (CHO) expressing different integrin subunits were tested for their capacity to bind the disintegrin domain of ADAMS. Mouse or human bone marrow cells and purified OCL precursors were tested for αvβ 1 integrin expression by Western blot, immunocytochemistry, and real-time RT-PCR. A monoclonal antibody to human α9 was used to block α9β1 on OCL precursors stimulated by 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D 3] or RANKL. Vertebrae of 7-day-old α9-/- mice and wildtype (WT) littermates were compared using bone histomorphometry and 3D μCT analysis. Results: α9 integrin was expressed by mouse and human bone marrow-derived OCLs and their precursors. Importantly, the anti-α9 antibody inhibited human OCL formation stimulated by 1α,25(OH)2D3 or RANKL dose-dependently. Furthermore, analysis of OCLs formed in marrow cultures from α9 -/- mice showed that the OCLs formed were more contracted and formed significantly less bone resorption pits on dentin slices. Histologic analysis of α9-/- vertebrae showed thickened trabecular regions and retained cartilage within vertebral bodies of α9 -/- mice. 3D μCT analysis of α9-/- vertebrae also showed a significant increase in trabecular bone volume/total tissue volume and a tendency for decreased trabecular separation compared with WT mice. Conclusions: These results support a previously unknown role for α9β1 integrin in OCL formation and function.

AB - We identified a previously unknown integrin, α9β 1, on OCLs and their precursors. Antibody to α9 inhibited OCL formation in human marrow cultures, and OCLs from α9 knockout mice had a defect in actin ring reorganization and an impaired bone resorption capacity. Introduction: Integrins play important roles in osteoclast (OCL) formation and function. Mature OCLs mainly express αvβ3 integrin, a heterodimer adhesion receptor that has been implicated in osteoclastic bone resorption. We identified ADAMS, a disintegrin and metalloproteinase, as a novel stimulator of OCL differentiation and showed that the disintegrin domain of ADAMS mediated its effects on OCL formation. Because the disintegrin domain of ADAMS does not bind Arg-Gly-Asp (RGD) sequences, we determined which integrin bound ADAMS and characterized its role in OCL formation and activity. Materials and Methods: Chinese hamster ovary cells (CHO) expressing different integrin subunits were tested for their capacity to bind the disintegrin domain of ADAMS. Mouse or human bone marrow cells and purified OCL precursors were tested for αvβ 1 integrin expression by Western blot, immunocytochemistry, and real-time RT-PCR. A monoclonal antibody to human α9 was used to block α9β1 on OCL precursors stimulated by 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D 3] or RANKL. Vertebrae of 7-day-old α9-/- mice and wildtype (WT) littermates were compared using bone histomorphometry and 3D μCT analysis. Results: α9 integrin was expressed by mouse and human bone marrow-derived OCLs and their precursors. Importantly, the anti-α9 antibody inhibited human OCL formation stimulated by 1α,25(OH)2D3 or RANKL dose-dependently. Furthermore, analysis of OCLs formed in marrow cultures from α9 -/- mice showed that the OCLs formed were more contracted and formed significantly less bone resorption pits on dentin slices. Histologic analysis of α9-/- vertebrae showed thickened trabecular regions and retained cartilage within vertebral bodies of α9 -/- mice. 3D μCT analysis of α9-/- vertebrae also showed a significant increase in trabecular bone volume/total tissue volume and a tendency for decreased trabecular separation compared with WT mice. Conclusions: These results support a previously unknown role for α9β1 integrin in OCL formation and function.

KW - α knockout mice

KW - αβ integrin

KW - ADAM8

KW - Osteoclast

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