The long term goal of this study is to define new therapeutic targets of osteoclast related bone diseases, such as osteoporosis and tumor bone metastasis. The immediate goal of this study is to understand the mechanisms underlying the transcription factors that regulate osteoclast lineage commitment and differentiation. The critical need for the treatments of osteoclast related bone diseases has been exacerbated by the recent discovery that the negative effects (e.g., jaw osteonecrosis) of currently used osteoclast inhibitors may outweigh the benefits. The mechanism underlying transcription factors that specify osteoclast lineage commitment, differentiation, and function remains unclear. It is also necessary to clarify why M-CSF alone induces precursors to differentiate into macrophages, while both M-CSF and RANKL induce precursors to differentiate into osteoclasts. Further study is needed to address these biological questions. As a step towards understanding the transcription factors which modulate osteoclast gene (e.g., cathepsin K) expression and osteoclast differentiation, we are characterizing the cathepsin K critical cis-regulatory elements (CCREs). We found a CCAAT/enhancer-binding protein ? (C/EBP?) binding site as a CCRE and confirmed C/EBP? as its trans-regulatory factor as the CCRE binding protein (CCREBP). Our Preliminary Studies further demonstrated that only the combined stimulation of M-CSF and RANKL will induce C/EBP? high expression;that in vitro C/EBP? knockdown blocks osteoclast differentiation and overexpression increase osteoclast formation;and that C/EBP? knockout in mice (C/EBP?-/-) impaired osteoclastogenesis and resulted in an osteopetrotic phenotype. Based on our Preliminary Study data, we hypothesize that C/EBP? is the key osteoclastogenesis transcription factor that regulates osteoclast lineage commitment and differentiation in the M-CSF and RANKL-dependent signaling pathway(s) through interactions with its heterodimerization partners and regulation of osteoclast genes. We will test this hypothesis through four specific aims:
in Aim 1, we will investigate the role of C/EBP? in osteoclast differentiation by characterizing the phenotypes and pathomechanism of C/EBP? null mutation mice (C/EBP?-/-) compared with C/EBP?+/+ mice.
In Aim 2, we will reveal the function of C/EBP? at various stages of osteoclast differentiation through characterization of the phenotypes and pathomechanism of three C/EBP? conditional knockout mouse models.
In Aim 3, we will define in vitro the roles of C/EBP? at different stages of osteoclast differentiation using RNAi knockdown as a loss-of-function study and retrovirus overexpression as a gain-of-function study.
In Aim 4, we dissect the mechanism of C/EBP? interactions that confers osteoclastic specificity by characterizing C/EBP? heterodimerization partners, downstream genes and their functions. This study will not only improve our understanding of the role of transcription factors in normal osteoclast differentiation and function and in osteolytic diseases (e.g., osteoporosis and bone metastases), but it will also facilitate the design of novel approaches for their precise treatment using drug or somatic gene therapy.

Public Health Relevance

This study endeavors to increase our understanding of the mechanisms underlying the transcription factors that regulate osteoclast lineage commitment and differentiation. In so doing, this study will elucidate how transcription factors specify osteoclast cell lineage commitment and differentiation. This study will not only improve our understanding of the role of transcription factors in normal osteoclast differentiation and functio and in osteolytic diseases (e.g., osteoporosis and bone metastases), but it will also facilitate th design of novel approaches for their precise treatment using drug or somatic gene therapy.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR044741-13
Application #
8641313
Study Section
Skeletal Biology Structure and Regeneration Study Section (SBSR)
Program Officer
Chen, Faye H
Project Start
1999-02-01
Project End
2017-03-31
Budget Start
2014-04-01
Budget End
2015-03-31
Support Year
13
Fiscal Year
2014
Total Cost
$424,059
Indirect Cost
$134,599
Name
University of Alabama Birmingham
Department
Pathology
Type
Schools of Medicine
DUNS #
063690705
City
Birmingham
State
AL
Country
United States
Zip Code
35294
Chen, Wei; Zhu, Guochun; Tang, Jun et al. (2018) C/ebp? controls osteoclast terminal differentiation, activation, function, and postnatal bone homeostasis through direct regulation of Nfatc1. J Pathol 244:271-282
Chen, Wei; Zhu, Guochun; Jules, Joel et al. (2018) Monocyte-Specific Knockout of C/ebp? Results in Osteopetrosis Phenotype, Blocks Bone Loss in Ovariectomized Mice, and Reveals an Important Function of C/ebp? in Osteoclast Differentiation and Function. J Bone Miner Res 33:691-703
Huang, Hong; Wang, Jue; Zhang, Yan et al. (2018) Bone resorption deficiency affects tooth root development in RANKL mutant mice due to attenuated IGF-1 signaling in radicular odontoblasts. Bone 114:161-171
Jules, Joel; Li, Yi-Ping; Chen, Wei (2018) C/EBP? and PU.1 exhibit different responses to RANK signaling for osteoclastogenesis. Bone 107:104-114
Jules, Joel; Chen, Wei; Feng, Xu et al. (2018) C/EBP? transcription factor is regulated by the RANK cytoplasmic 535IVVY538 motif and stimulates osteoclastogenesis more strongly than c-Fos. J Biol Chem 293:1480-1492
Wu, Mengrui; Chen, Wei; Lu, Yun et al. (2017) G?13 negatively controls osteoclastogenesis through inhibition of the Akt-GSK3?-NFATc1 signalling pathway. Nat Commun 8:13700
Pan, Jie; Wang, Jue; Hao, Liang et al. (2017) The Triple Functions of D2 Silencing in Treatment of Periapical Disease. J Endod 43:272-278
McConnell, Matthew; Feng, Shengmei; Chen, Wei et al. (2017) Osteoclast proton pump regulator Atp6v1c1 enhances breast cancer growth by activating the mTORC1 pathway and bone metastasis by increasing V-ATPase activity. Oncotarget 8:47675-47690
Wu, Mengrui; Wang, Yiping; Shao, Jian-Zhong et al. (2017) Cbf? governs osteoblast-adipocyte lineage commitment through enhancing ?-catenin signaling and suppressing adipogenesis gene expression. Proc Natl Acad Sci U S A 114:10119-10124
Jules, Joel; Chen, Wei; Feng, Xu et al. (2016) CCAAT/Enhancer-binding Protein ? (C/EBP?) Is Important for Osteoclast Differentiation and Activity. J Biol Chem 291:16390-403

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