Our long-term goal is to understand the molecular mechanisms regulating growth plate development. The last step in maturation of the growth plate is the differentiation of proliferative chondrocytes into hypertrophic chondrocytes, which subsequently undergo endochondral ossification. Our recent findings indicate that histone deacetylation 4 (HDAC4) nuclear-cytoplasmic shuttling, degradation, and translational repression plays a major role in this process. Specifically, we suspect that HDAC4 functions as a negative regulator of chondrocyte hypertrophy by binding and inhibiting Runx 2/Cbfa1 expression in the nucleus. Runx 2/Cbfa1 is a transcription factor necessary for chondrocyte differentiation and hypertrophy. However, the mechanisms underlying HDAC4 shuttling, degradation, and translational repression are as yet undetermined. Our overall hypothesis is that the relocation, degradation, and translational repression of HDAC4 control chondrocyte differentiation and are regulated by Ca2+/calmodulin kinase IV (CaMKIV), p38 MAP kinase, and microRNA-1 respectively at different zone in the growth plate. The hypothesis includes three specific aims:
Specific Aim 1 : To determine whether activation of the Ca2+/calmodulin signaling pathway prevents nuclear entry of HDAC4 and enhances the binding of HDAC4 to the cytoplasmic binding protein 14-3-3. This may impair HDAC4-mediated inhibition of chondrocyte differentiation in the nucleus. Hypothesis 1 is that HDAC4 nuclear- cytoplasmic shuttling controls chondrocyte differentiation and is dependent on the Ca2+/calmodulin signaling pathway.
Specific Aim 2 : To determine whether: 2a) caspases degrade HDAC4;2b) p38 MAPK regulates expression or activity of capsases 2 and 3;2c) Runx2 expression is dependent on p38 MAPK by in situ hybridization in p38 MAPK defective and constitutively activated MKK6 mouse growth plates. Project 2b will be tested using active MKK6 to elevate p38 and dominant negative p38 MAPK to repress p38. Hypothesis 2 is that HDAC4 degradation is regulated by P38 MAPK by increasing expression of caspases 2 and 3.
Specific Aim 3 : To determine whether: 3a) the spatio-temporal distribution of specific miRNA-1 is different during growth plate development;3b) miRNA-1 downregulates HDAC4 by repressing HDAC4 translation at 3'UTR in the chondrocytes;3c) miRNA-1regulates chondrocyte proliferation and differentiation. Hypothesis 3 is that microRNA-1 is involved in chondrocyte hypertrophy regulation by repressing HDAC4 translation. Significance of the growth plate, or physis, determines the future length and shape of the human mature bone. Delineating the physiological controls of the physis could suggest possibilities for therapeutically manipulating endochondral bone growth by modulating the signaling pathways that govern the association of HDAC4 with Runx2. Prevention of leg length discrepancy, dwarfism, and other disorders of bone growth might also be possible once molecular triggers and stops affecting the growth plate are better understood.

Public Health Relevance

Although many factors have been found to impact the function of the growth plate, its biology remains poorly understood. The importance of clearly delineating the physiological controls of the physis by histone deacetylation cannot be overemphasized. Such knowledge could allow the prevention or correction of a number of bone-growth related problems in adults and children. Leg length discrepancy and dwarfism are just a few clinical dilemmas that may be provided with more elegant solutions if the molecular triggers and stops to the growth plate can be elucidated. The understanding of cell differentiation in growth plate may also be important to the understanding of abnormal chondrocyte differentiation in osteoarthritis.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01AR059142-04
Application #
8635976
Study Section
Skeletal Biology Structure and Regeneration Study Section (SBSR)
Program Officer
Tyree, Bernadette
Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Rhode Island Hospital
Department
Type
DUNS #
City
Providence
State
RI
Country
United States
Zip Code
02903
Li, Pengcui; Deng, Jin; Wei, Xiaochun et al. (2016) Blockade of hypoxia-induced CXCR4 with AMD3100 inhibits production of OA-associated catabolic mediators IL-1β and MMP-13. Mol Med Rep 14:1475-82
Chen, Chongwei; Wei, Xiaochun; Wang, Shaowei et al. (2016) Compression regulates gene expression of chondrocytes through HDAC4 nuclear relocation via PP2A-dependent HDAC4 dephosphorylation. Biochim Biophys Acta 1863:1633-42
Du, Guoqing; Zhan, Hongsheng; Ding, Daofang et al. (2016) Abnormal Mechanical Loading Induces Cartilage Degeneration by Accelerating Meniscus Hypertrophy and Mineralization After ACL Injuries In Vivo. Am J Sports Med 44:652-63
Li, Yong Ping; Wei, Xiao Chun; Li, Peng Cu et al. (2015) The Role of miRNAs in Cartilage Homeostasis. Curr Genomics 16:393-404
Zhou, Jingming; Li, Pengcui; Chen, Qian et al. (2015) Mitogen-activated protein kinase p38 induces HDAC4 degradation in hypertrophic chondrocytes. Biochim Biophys Acta 1853:370-6
Thomas, Nathan P; Li, Pengcui; Fleming, Braden C et al. (2015) Attenuation of cartilage pathogenesis in post-traumatic osteoarthritis (PTOA) in mice by blocking the stromal derived factor 1 receptor (CXCR4) with the specific inhibitor, AMD3100. J Orthop Res 33:1071-8
Sun, Xiaojuan; Charbonneau, Cherie; Wei, Lei et al. (2015) miR-181a Targets RGS16 to Promote Chondrosarcoma Growth, Angiogenesis, and Metastasis. Mol Cancer Res 13:1347-57
Wang, Shaowei; Zhou, Jingming; Wei, Xiaochun et al. (2015) Identification of α1-Antitrypsin as a Potential Candidate for Internal Control for Human Synovial Fluid in Western Blot. Rheumatology (Sunnyvale) Suppl 6:006
Cao, Kun; Wei, Lei; Zhang, Zhiqiang et al. (2014) Decreased histone deacetylase 4 is associated with human osteoarthritis cartilage degeneration by releasing histone deacetylase 4 inhibition of runt-related transcription factor-2 and increasing osteoarthritis-related genes: a novel mechanism of human ost Arthritis Res Ther 16:491
Zhou, Jingming; Chen, Qian; Lanske, Beate et al. (2014) Disrupting the Indian hedgehog signaling pathway in vivo attenuates surgically induced osteoarthritis progression in Col2a1-CreERT2; Ihhfl/fl mice. Arthritis Res Ther 16:R11

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