Abstract
The LIM homeobox 2 (Lhx2) transcription factor Lhx2 has a variety of functions, including neural induction, morphogenesis, and hematopoiesis. Here we show the involvement of Lhx2 in osteoclast differentiation. Lhx2 was strongly expressed in osteoclast precursor cells but its expression was significantly reduced during receptor activator of nuclear factor-κB ligand (RANKL)-mediated osteoclastogenesis. Overexpression of Lhx2 in bone marrow-derived monocyte/macrophage lineage cells (BMMs), which are osteoclast precursor cells, attenuated RANKL-induced osteoclast differentiation by inhibiting the induction of nuclear factor of activated T cells c1 (NFATc1). Interestingly, interaction of Lhx2 proteins with c-Fos attenuated the DNA-binding ability of c-Fos and thereby inhibited the transactivation of NFATc1. Furthermore, Lhx2 conditional knockout mice exhibited an osteoporotic bone phenotype, which was related with increased osteoclast formation in vivo. Taken together, our results suggest that Lhx2 acts as a negative regulator of osteoclast formation in vitro and in vivo. The anti-osteoclastogenic effect of Lhx2 may be useful for developing a therapeutic strategy for bone disease.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- RANKL:
-
receptor activator of nuclear factor-κB ligand
- 1,25(OH)2D3:
-
1α,25-dihydroxyvitamin D3
- BMP:
-
bone morphogenetic protein
- ALP:
-
alkaline phosphatase
- TRAP:
-
tartrate-resistant acid phosphate
- M-CSF:
-
macrophage colony-stimulating factor
- JNK:
-
c-Jun N-terminal kinase
- ERK:
-
extracellular signal-regulated kinase
- NFAT:
-
nuclear factor of activated T cells
- TNF:
-
tumor necrosis factor
- IFN:
-
interferon
- IL-6:
-
interleukin-6
- OSCAR:
-
osteoclast-associated receptor
- OPG:
-
osteoprotegerin
- BMM:
-
bone marrow-derived macrophage lineage cell
- ChIP:
-
chromatin immunoprecipitation
- Lhx2:
-
LIM homeobox 2
- NFATc1:
-
nuclear factor of activated T cells c1
- MNC:
-
multinuclear cell
- EMSA:
-
electrophoretic mobility shift assay
References
Walsh MC, Kim N, Kadono Y, Rho J, Lee SY, Lorenzo J et al. Osteoimmunology: interplay between the immune system and bone metabolism. Annu Rev Immunol 2006; 24: 33–63.
Agata H, Asahina I, Yamazaki Y, Uchida M, Shinohara Y, Honda MJ et al. Effective bone engineering with periosteum-derived cells. J Dent Res 2007; 86: 79–83.
Arai F, Miyamoto T, Ohneda O, Inada T, Sudo T, Brasel K et al. Commitment and differentiation of osteoclast precursor cells by the sequential expression of c-Fms and receptor activator of nuclear factor kappaB (RANK) receptors. J Exp Med 1999; 190: 1741–1754.
Takayanagi H, Kim S, Koga T, Nishina H, Isshiki M, Yoshida H et al. Induction and activation of the transcription factor NFATc1 (NFAT2) integrate RANKL signaling in terminal differentiation of osteoclasts. Dev Cell 2002; 3: 889–901.
Grigoriadis AE, Wang ZQ, Cecchini MG, Hofstetter W, Felix R, Fleisch HA et al. c-Fos: a key regulator of osteoclast-macrophage lineage determination and bone remodeling. Science 1994; 266: 443–448.
Matsuo K, Galson DL, Zhao C, Peng L, Laplace C, Wang KZ et al. Nuclear factor of activated T-cells (NFAT) rescues osteoclastogenesis in precursors lacking c-Fos. J Biol Chem 2004; 279: 26475–26480.
Kim K, Kim JH, Lee J, Jin HM, Lee SH, Fisher DE et al. Nuclear factor of activated T cells c1 induces osteoclast-associated receptor gene expression during tumor necrosis factor-related activation-induced cytokine-mediated osteoclastogenesis. J Biol Chem 2005; 280: 35209–35216.
Matsumoto M, Kogawa M, Wada S, Takayanagi H, Tsujimoto M, Katayama S et al. Essential role of p38 mitogen-activated protein kinase in cathepsin K gene expression during osteoclastogenesis through association of NFATc1 and PU.1. J Biol Chem 2004; 279: 45969–45979.
Aliprantis AO, Ueki Y, Sulyanto R, Park A, Sigrist KS, Sharma SM et al. NFATc1 in mice represses osteoprotegerin during osteoclastogenesis and dissociates systemic osteopenia from inflammation in cherubism. J Clin Invest 2008; 118: 3775–3789.
Dawid IB, Breen JJ, Toyama R . LIM domains: multiple roles as adapters and functional modifiers in protein interactions. Trends Genet 1998; 14: 156–162.
Bach I, Rhodes SJ, Pearse RV 2nd, Heinzel T, Gloss B, Scully KM et al. P-Lim, a LIM homeodomain factor, is expressed during pituitary organ and cell commitment and synergizes with Pit-1. Proc Natl Acad Sci USA 1995; 92: 2720–2724.
Xu Y, Baldassare M, Fisher P, Rathbun G, Oltz EM, Yancopoulos GD et al. LH-2: a LIM/homeodomain gene expressed in developing lymphocytes and neural cells. Proc Natl Acad Sci USA 1993; 90: 227–231.
Porter FD, Drago J, Xu Y, Cheema SS, Wassif C, Huang SP et al. Lhx2, a LIM homeobox gene, is required for eye, forebrain, and definitive erythrocyte development. Development 1997; 124: 2935–2944.
Tetreault N, Champagne MP, Bernier G . The LIM homeobox transcription factor Lhx2 is required to specify the retina field and synergistically cooperates with Pax6 for Six6 trans-activation. Dev Biol 2009; 327: 541–550.
Tornqvist G, Sandberg A, Hagglund AC, Carlsson L . Cyclic expression of lhx2 regulates hair formation. PLoS Genet 2010; 6: e1000904.
Hirota J, Mombaerts P . The LIM-homeodomain protein Lhx2 is required for complete development of mouse olfactory sensory neurons. Proc Natl Acad Sci USA 2004; 101: 8751–8755.
Lee SK, Kalinowski J, Jastrzebski S, Lorenzo JA . 1,25(OH)2 vitamin D3-stimulated osteoclast formation in spleen-osteoblast cocultures is mediated in part by enhanced IL-1 alpha and receptor activator of NF-kappa B ligand production in osteoblasts. J Immunol 2002; 169: 2374–2380.
Suda T, Takahashi N, Udagawa N, Jimi E, Gillespie MT, Martin TJ . Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. Endocrine Rev 1999; 20: 345–357.
Hobert O, Westphal H . Functions of LIM-homeobox genes. Trends Genet 2000; 16: 75–83.
Kim KK, Song SB, Kang KI, Rhee M, Kim KE . Activation of the thyroid-stimulating hormone beta-subunit gene by LIM homeodomain transcription factor Lhx2. Endocrinology 2007; 148: 3468–3476.
Hou PS, Chuang CY, Kao CF, Chou SJ, Stone L, Ho HN et al. LHX2 regulates the neural differentiation of human embryonic stem cells via transcriptional modulation of PAX6 and CER1. Nucleic Acids Res 2013; 41: 7753–7770.
Glenn DJ, Maurer RA . MRG1 binds to the LIM domain of Lhx2 and may function as a coactivator to stimulate glycoprotein hormone alpha-subunit gene expression. J Biol Chem 1999; 274: 36159–36167.
Perez C, Dastot-Le Moal F, Collot N, Legendre M, Abadie I, Bertrand AM et al. Screening of LHX2 in patients presenting growth retardation with posterior pituitary and ocular abnormalities. Eur J Endocrinol 2012; 167: 85–91.
Mangale VS, Hirokawa KE, Satyaki PR, Gokulchandran N, Chikbire S, Subramanian L et al. Lhx2 selector activity specifies cortical identity and suppresses hippocampal organizer fate. Science 2008; 319: 304–309.
Lian JB, Stein GS, Javed A, van Wijnen AJ, Stein JL, Montecino M et al. Networks and hubs for the transcriptional control of osteoblastogenesis. Rev Endocr Metab Disord 2006; 7: 1–16.
Kaneko H, Arakawa T, Mano H, Kaneda T, Ogasawara A, Nakagawa M et al. Direct stimulation of osteoclastic bone resorption by bone morphogenetic protein (BMP)-2 and expression of BMP receptors in mature osteoclasts. Bone 2000; 27: 479–486.
Kim K, Kim JH, Lee J, Jin HM, Kook H, Kim KK et al. MafB negatively regulates RANKL-mediated osteoclast differentiation. Blood 2007; 109: 3253–3259.
Lee J, Kim K, Kim JH, Jin HM, Choi HK, Lee SH et al. Id helix-loop-helix proteins negatively regulate TRANCE-mediated osteoclast differentiation. Blood 2006; 107: 2686–2693.
Zhao B, Takami M, Yamada A, Wang X, Koga T, Hu X et al. Interferon regulatory factor-8 regulates bone metabolism by suppressing osteoclastogenesis. Nat Med 2009; 15: 1066–1071.
Kobayashi K, Takahashi N, Jimi E, Udagawa N, Takami M, Kotake S et al. Tumor necrosis factor alpha stimulates osteoclast differentiation by a mechanism independent of the ODF/RANKL-RANK interaction. J Exp Med 2000; 191: 275–286.
Queiroz-Junior CM, Bessoni RL, Costa VV, Souza DG, Teixeira MM, Silva TA . Preventive and therapeutic anti-TNF-alpha therapy with pentoxifylline decreases arthritis and the associated periodontal co-morbidity in mice. Life Sci 2013; 93: 423–428.
Chou SJ, Perez-Garcia CG, Kroll TT, O'Leary DD . Lhx2 specifies regional fate in Emx1 lineage of telencephalic progenitors generating cerebral cortex. Nat Neurosci 2009; 12: 1381–1389.
Parfitt AM, Drezner MK, Glorieux FH, Kanis JA, Malluche H, Meunier PJ et al. Bone histomorphometry: standardization of nomenclature, symbols, and units. Report of the ASBMR Histomorphometry Nomenclature Committee. J Bone Miner Res 1987; 2: 595–610.
Acknowledgements
This work was supported by the National Research Foundation of Korea (NRF) grant (MRC for Gene Regulation, 2011-0030132) funded by the Korea government (MSIP) and a Grant A110703 of the Korean Health Technology R&D Project, Ministry of Health and Welfare (to JK).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Edited by JP Medema
Supplementary Information accompanies this paper on Cell Death and Differentiation website
Supplementary information
Rights and permissions
About this article
Cite this article
Kim, J., Youn, B., Kim, K. et al. Lhx2 regulates bone remodeling in mice by modulating RANKL signaling in osteoclasts. Cell Death Differ 21, 1613–1621 (2014). https://doi.org/10.1038/cdd.2014.71
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/cdd.2014.71
This article is cited by
-
Recent advances of NFATc1 in rheumatoid arthritis-related bone destruction: mechanisms and potential therapeutic targets
Molecular Medicine (2024)
-
LHX2 facilitates the progression of nasopharyngeal carcinoma via activation of the FGF1/FGFR axis
British Journal of Cancer (2022)
-
Selenoprotein W ensures physiological bone remodeling by preventing hyperactivity of osteoclasts
Nature Communications (2021)
-
Genomic imbalances defining novel intellectual disability associated loci
Orphanet Journal of Rare Diseases (2019)
-
mTORC1 impedes osteoclast differentiation via calcineurin and NFATc1
Communications Biology (2018)
