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J. Biol. Chem., Vol. 278, Issue 46, 45368-45374, November 14, 2003

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₁ Integrin/Focal Adhesion Kinase-mediated Signaling Induces Intercellular Adhesion Molecule 1 and Receptor Activator of Nuclear Factor B Ligand on Osteoblasts and Osteoclast Maturation^*

Shingo Nakayamada, Yosuke Okada, Kazuyoshi Saito, Masahito Tamura, and Yoshiya Tanaka¶

From the First and Second Department of Internal Medicine, University of Occupational and Environmental Health, Japan, School of Medicine, Kitakyushu, 807-8555, Japan

We have assessed characteristics of primary human osteoblasts, shedding light on signaling mediated by ₁ integrin. ₁ integrins are major receptors for these matrix glycoproteins. 1) Integrins ₁, ₂, ₃, ₄, ₅, ₆, and _v were highly expressed on primary osteoblasts. 2) Engagement of ₁ integrins on osteoblasts by cross-linking with specific antibody or ligand matrices, such as fibronectin or collagen, augmented expression of intercellular adhesion molecule 1 (ICAM-1) and receptor activator of nuclear factor B ligand (RANKL) on the surface. 3) Up-regulation of ICAM-1 and RANKL on osteoblasts by ₁ stimulation was completely abrogated by pretreatment with herbimycin A and genistein, tyrosine kinase inhibitors, or transfection of dominant negative truncations of focal adhesion kinase (FAK). 4) Engagement of ₁ integrins on osteoblasts induced tartrate-resistant acid phosphatase-positive multinuclear cell formation in the coculture system of osteoblasts and peripheral monocytes. 5) Up-regulation of tartrate-resistant acid phosphatase-positive multinuclear cell formation by ₁ stimulation was completely abrogated by transfection of dominant negative truncations of FAK. Our results indicate that ₁ integrin-dependent adhesion of osteoblasts to bone matrices induces ICAM-1 and RANKL expression and osteoclast formation via tyrosine kinase, especially FAK. We here propose that ₁ integrin/FAK-mediated signaling on osteoblasts could be involved in ICAM-1- and RANKL-dependent osteoclast maturation.

Received for publication, August 8, 2003 , and in revised form, September 2, 2003.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom correspondence should be addressed. Tel.: 81-93-603-1611; Fax: 81-93-691-9334; E-mail: tanaka{at}med.uoeh-u.ac.jp.

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