Focal Adhesion Kinase Signaling Pathways Regulate the Osteogenic Differentiation of Human Mesenchymal Stem Cells

Overview

Journal Exp Cell Res

Specialty Cell Biology

Date 2006 Nov 4

PMID 17081517

Citations 110

Authors

Roman M Salasznyk

Robert F Klees

William A Williams

Adele Boskey

George E Plopper

Affiliations

Soon will be listed here.

Abstract

The intracellular signaling events controlling human mesenchymal stem cells (hMSC) differentiation into osteoblasts are not entirely understood. We recently demonstrated that contact with extracellular matrix (ECM) proteins is sufficient to induce osteogenic differentiation of hMSC through an ERK-dependent pathway. We hypothesized that FAK signaling pathways provide a link between activation of ERK1/2 by ECM, and stimulate subsequent phosphorylation of the Runx2/Cbfa-1 transcription factor that controls osteogenic gene expression. We plated hMSC on purified collagen I (COLL-I) and vitronectin (VN) in the presence or absence of FAK-specific siRNA, and assayed for phosphorylation of Runx2/Cbfa-1 as well as expression of established osteogenic differentiation markers (bone sialoprotein-2, osteocalcin, alkaline phosphatase, calcium deposition, and spectroscopically determined mineral:matrix ratio). We found that siRNA treatment reduced FAK mRNA levels by >40% and decreased ECM-mediated phosphorylation of FAK Y397 and ERK1/2. Serine phosphorylation of Runx2/Cbfa-1 was significantly reduced after 8 days in treated cells. Finally, FAK inhibition blocked osterix transcriptional activity and the osteogenic differentiation of hMSC, as assessed by lowered expression of osteogenic genes (RT-PCR), decreased alkaline phosphatase activity, greatly reduced calcium deposition, and a lower mineral:matrix ratio after 28 days in culture. These results suggest that FAK signaling plays an important role in regulating ECM-induced osteogenic differentiation of hMSC.

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References

Yamada K, Green K, Samarel A, Saffitz J . Distinct pathways regulate expression of cardiac electrical and mechanical junction proteins in response to stretch. Circ Res. 2005; 97(4):346-53. DOI: 10.1161/01.RES.0000178788.76568.8a. View

Salasznyk R, Klees R, Hughlock M, Plopper G . ERK signaling pathways regulate the osteogenic differentiation of human mesenchymal stem cells on collagen I and vitronectin. Cell Commun Adhes. 2005; 11(5-6):137-53. DOI: 10.1080/15419060500242836. View

Simmons C, Matlis S, Thornton A, Chen S, Wang C, Mooney D . Cyclic strain enhances matrix mineralization by adult human mesenchymal stem cells via the extracellular signal-regulated kinase (ERK1/2) signaling pathway. J Biomech. 2003; 36(8):1087-96. DOI: 10.1016/s0021-9290(03)00110-6. View

Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K . Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell. 1997; 89(5):755-64. DOI: 10.1016/s0092-8674(00)80258-5. View

Schaller M . Biochemical signals and biological responses elicited by the focal adhesion kinase. Biochim Biophys Acta. 2001; 1540(1):1-21. DOI: 10.1016/s0167-4889(01)00123-9. View

Plopper G, Domanico S, Cirulli V, Kiosses W, Quaranta V . Migration of breast epithelial cells on Laminin-5: differential role of integrins in normal and transformed cell types. Breast Cancer Res Treat. 1999; 51(1):57-69. DOI: 10.1023/a:1006086218174. View

Jaiswal R, Jaiswal N, Bruder S, Mbalaviele G, Marshak D, Pittenger M . Adult human mesenchymal stem cell differentiation to the osteogenic or adipogenic lineage is regulated by mitogen-activated protein kinase. J Biol Chem. 2000; 275(13):9645-52. DOI: 10.1074/jbc.275.13.9645. View

Guo Z, Liu X, Hou C, Li X, Mao N . [Human Bone Marrow Mesenchymal Stem Cells Differentiate into Neuron-Like Cells In Vitro]. Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2003; 9(1):91-92. View

Xiao G, Wang D, Benson M, Karsenty G, Franceschi R . Role of the alpha2-integrin in osteoblast-specific gene expression and activation of the Osf2 transcription factor. J Biol Chem. 1998; 273(49):32988-94. DOI: 10.1074/jbc.273.49.32988. View

10.

Salasznyk R, Klees R, Westcott A, Vandenberg S, Bennett K, Plopper G . Focusing of gene expression as the basis of stem cell differentiation. Stem Cells Dev. 2006; 14(6):608-20. DOI: 10.1089/scd.2005.14.608. View

11.

Xiao G, Jiang D, Thomas P, Benson M, Guan K, Karsenty G . MAPK pathways activate and phosphorylate the osteoblast-specific transcription factor, Cbfa1. J Biol Chem. 2000; 275(6):4453-9. DOI: 10.1074/jbc.275.6.4453. View

12.

Klees R, Salasznyk R, Kingsley K, Williams W, Boskey A, Plopper G . Laminin-5 induces osteogenic gene expression in human mesenchymal stem cells through an ERK-dependent pathway. Mol Biol Cell. 2004; 16(2):881-90. PMC: 545919. DOI: 10.1091/mbc.e04-08-0695. View

13.

Essayem S, Kovacic-Milivojevic B, Baumbusch C, McDonagh S, Dolganov G, Howerton K . Hair cycle and wound healing in mice with a keratinocyte-restricted deletion of FAK. Oncogene. 2005; 25(7):1081-9. PMC: 2710133. DOI: 10.1038/sj.onc.1209130. View

14.

Perinpanayagam H, Zaharias R, Stanford C, Brand R, Keller J, Schneider G . Early cell adhesion events differ between osteoporotic and non-osteoporotic osteoblasts. J Orthop Res. 2002; 19(6):993-1000. DOI: 10.1016/S0736-0266(01)00045-6. View

15.

Franceschi R, Xiao G . Regulation of the osteoblast-specific transcription factor, Runx2: responsiveness to multiple signal transduction pathways. J Cell Biochem. 2003; 88(3):446-54. DOI: 10.1002/jcb.10369. View

16.

Burridge K, Chrzanowska-Wodnicka M . Focal adhesions, contractility, and signaling. Annu Rev Cell Dev Biol. 1996; 12:463-518. DOI: 10.1146/annurev.cellbio.12.1.463. View

17.

Koukouritaki S, Lianos E . Glucocorticoid effect on human mesangial cell cytoskeletal proteins. J Lab Clin Med. 1999; 133(4):378-83. DOI: 10.1016/s0022-2143(99)90069-0. View

18.

Otto F, Thornell A, Crompton T, Denzel A, Gilmour K, Rosewell I . Cbfa1, a candidate gene for cleidocranial dysplasia syndrome, is essential for osteoblast differentiation and bone development. Cell. 1997; 89(5):765-71. DOI: 10.1016/s0092-8674(00)80259-7. View

19.

Salasznyk R, Westcott A, Klees R, Ward D, Xiang Z, Vandenberg S . Comparing the protein expression profiles of human mesenchymal stem cells and human osteoblasts using gene ontologies. Stem Cells Dev. 2005; 14(4):354-66. DOI: 10.1089/scd.2005.14.354. View

20.

McLean G, Komiyama N, Serrels B, Asano H, Reynolds L, Conti F . Specific deletion of focal adhesion kinase suppresses tumor formation and blocks malignant progression. Genes Dev. 2004; 18(24):2998-3003. PMC: 535910. DOI: 10.1101/gad.316304. View