HIPPO SIGNALING PROTEIN MST1 REGULATES OSTEOCLAST DIFFERENTIATION BY INTERACTING WITH INTEGRIN LINKED KINASE (ILK) AND MODULATING ACTIN STRUCTURES
Abstract
Hippo signaling is implicated in balancing cell proliferation, differentiation and death in multiple organs. However, its role in specific bone cell types such as osteoclasts, and its significance in maintaining overall bone tissue homeostasis remain largely unknown. In this study, we investigated the role of the Hippo pathway in osteoclast differentiation. Human primary monocyte cells were treated with receptor activator nuclear factor kappaB ligand (RANKL) and evaluated for osteoclast differentiation by marker protein analysis, tartrate-resistant acid phosphate (TRAP) and resorption assays. Our results showed that Ste20-like kinase 1 (MST1) underwent the maximum change after RANKL treatments and is negatively associated with osteoclast differentiation. Furthermore, proteomic approaches involving co-immunoprecipitation and mass spectrometry identified MST1 interaction with integrin-linked kinase (ILK) which is lost during RANKL induced differentiation. Finally, using RNAi and ectopic expression experiments we observed that MST1-ILK interaction negatively inhibits osteoclast differentiation at the level of actin ring structure formation, which is facilitated by ILK. Together, our data highlight a role for the Hippo pathway protein, MST1, in negatively regulating osteoclast differentiation through its interaction with integrin signaling. Given that integrin signaling is progressively implicated in pathological osteolysis, augmenting this pathway could have therapeutic implications.
Key words: osteoclast; Hippo signaling; Ste20-like kinase 1 (MST1); integrin linked kinase (ILK); integrin signaling; actin ring structures
Received: September 29, 2015; Revised: November 15, 2015; Accepted: November 25, 2015
Downloads
References
Yu FX, Guan KL. The Hippo pathway: regulators and regulations. Genes Dev. 2013;27(4):355-71.
Staley BK, Irvine KD. Hippo signaling in Drosophila: recent advances and insights. Dev Dyn. 2012;241(1):3-15.
Harvey KF, Zhang X, Thomas DM. The Hippo pathway and human cancer. Nat Rev Cancer. 2013;13:246-57.
Hong W, Guan KL. The YAP and TAZ transcription co-activators: key downstream effectors of the mammalian Hippo pathway. Semin Cell Dev Biol. 2012;23(7):785-93.
Hansen CG, Moroishi T, Guan KL. YAP and TAZ: a nexus for Hippo signaling and beyond. Trends Cell Biol. 2015;25(9):499-513.
Hergovich A. Regulation and functions of mammalian LATS/NDR kinases: looking beyond canonical Hippo signalling. Cell Biosci. 2013;3(1):32.
Hergovich A, Stegert MR, Schmitz D, Hemmings BA. NDR kinases regulate essential cell processes from yeast to humans. Nat Rev Mol Cell Biol. 2006;7(4):253-64.
Boyle WJ, Simonet WS, Lacey DL. Osteoclast differentiation and activation. Nature. 2003;423(6937):337-42.
Teitelbaum SL. Bone resorption by osteoclasts. Science. 2000;289(5484):1504-8.
Teitelbaum SL Osteoclasts, integrins, and osteoporosis. J Bone Miner Metab. 2000;18(6):344-9.
Ikeda K, Takeshita S. Factors and mechanisms involved in the coupling from bone resorption to formation: how osteoclasts talk to osteoblasts. J Bone Metab. 2014;21(3):163-7.
Lee SK, Lorenzo J. Cytokines regulating osteoclast formation and function. Curr Opin Rheumatol. 2006;18(4):411-8.
Mabey T, Honsawek S. Cytokines as biochemical markers for knee osteoarthritis. World J Orthop. 2015;6(1):95-105.
Marks SC Jr. Osteoclast biology: lessons from mammalian mutations. Am. J. Med. Genet. 1989;34(1):43-53.
McLean W, Olsen BR. Mouse models of abnormal skeletal development and homeostasis. Trends Genet. 2001;17(10):S38-S43.
Hayden RS, Fortin JP, Harwood B, Subramanian B, Quinn KP, Georgakoudi I, Kopin AS, Kaplan DL. Cell-tethered ligands modulate bone remodeling by osteoblasts and osteoclasts. Adv Funct Mater. 2014;24(4):472-9.
Sørensen MG, Henriksen K, Schaller S, Henriksen DB, Nielsen FC, Dziegiel MH, Karsdal MA. Characterization of osteoclasts derived from CD14+ monocytes isolated from peripheral blood. J Bone Miner Metab. 2007;25:36-45.
Yoshida CA, Komori H, Maruyama Z, Miyazaki T, Kawasaki K, Furuichi T, Fukuyama R, Mori M, Yamana K, Nakamura K, Liu W, Toyosawa S, Moriishi T, Kawaguchi H, Takada K, Komori T. SP7 inhibits osteoblast differentiation at a late stage in mice. PLoS One. 2012;7(3):e32364.
Subramanian B, Ko WC, Yadav V, DesRochers TM, Perrone RD, Zhou J, Kaplan DL. The regulation of cystogenesis in a tissue engineered kidney disease system by abnormal matrix interactions. Biomaterials. 2012;33(33):8383-94.
Lee J, Youn BU, Kim K, Kim JH, Lee DH, Seong S, Kim I, Han SH, Che X, Choi JY, Park YW, Kook H, Kim KK, Lim DS, Kim N. Mst2 Controls Bone Homeostasis by Regulating Osteoclast and Osteoblast Differentiation. J Bone Miner Res. 2015;30(9):1597-607.
Dossa T, Arabian A, Windle JJ, Dedhar S, Teitelbaum SL, Ross FP, Roodman GD, St-Arnaud R. Osteoclast-specific inactivation of the integrin-linked kinase (ILK) inhibits bone resorption. J Cell Biochem. 2010;110(4):960-7.
Qian Y, Zhong X, Flynn DC, Zheng JZ, Qiao M, Wu C, Dedhar S, Shi X, Jiang BH. ILK mediates actin filament rearrangements and cell migration and invasion through PI3K/Akt/Rac1 signaling. Oncogene. 2005;24(19):3154-65.
Takahashi N, Udagawa N, Suda T. A new member of tumor necrosis factor ligand family, ODF/OPGL/TRANCE/RANKL, regulates osteoclast differentiation and function. Biochem Biophys Res Commun. 1999;256(3):449-55.
Takeshita S, Kaji K, Kudo A. Identification and characterization of the new osteoclast progenitor with macrophage phenotypes being able to differentiate into mature osteoclasts. J Bone Miner Res. 2000;15(8):1477-88.
Roux S, Orcel P. Bone loss. Factors that regulate osteoclast differentiation: an update. Arthritis Res. 2000;2(6):451-6.
Duong LT, Lakkakorpi P, Nakamura I, Rodan GA. Integrins and signaling in osteoclast function. Matrix Biol. 2000;19(2):97-105.
Zou W, Teitelbaum SL. Integrins, growth factors, and the osteoclast cytoskeleton. Ann N Y Acad Sci. 2010;1192:27–31.
Nesbitt S, Nesbit A, Helfrich M, Horton M. Biochemical characterization of human osteoclast integrins. Osteoclasts express alpha v beta 3, alpha 2 beta 1, and alpha v beta 1 integrins. J Biol Chem. 1993;268(22):16737-45.
Dedhar S, Williams B, Hannigan G. Integrin-linked kinase (ILK): a regulator of integrin and growth-factor signalling. Trends Cell Biol. 1999;9(8):319-23.
Teitelbaum SL. The osteoclast and its unique cytoskeleton. Ann N Y Acad Sci. 2011;1240:14-7.
Matsubara T, Myoui A, Ikeda F, Hata K, Yoshikawa H, Nishimura R, Yoneda T. Critical role of cortactin in actin ring formation and osteoclastic bone resorption. J Bone Miner Metab. 2006;24(5):368-72.
Lakkakorpi PT, Väänänen HK. Cytoskeletal changes in osteoclasts during the resorption cycle. Microsc Res Tech. 1996;33(2):171-81.
Downloads
Published
How to Cite
Issue
Section
License
Authors grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution 4.0 International License that allows others to share the work with an acknowledgment of the work’s authorship and initial publication in this journal.
