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Chaperonin 60 regulation of SOX9 ubiquitination mitigates the development of knee osteoarthritis

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Abstract

Articular cartilage integrity loss is a prominently deleterious feature of osteoarthritis (OA). The mechanistic underlying the development of OA warrants characterization. Heat shock proteins (HSPs), members of the chaperone family, reportedly orchestrate tissue homeostasis and remodeling in response to detrimental stress. This study was undertaken to characterize the biological role of HSP60 in the pathogenesis of OA knee. Articular specimens from OA knee patients displayed severe articular damage histopathology concomitant with low HSP60 concentrations in cartilage and synovial fluid compared to non-OA patients. In vitro, a gain of HSP60 signaling counteracted the IL-1β-mediated suppression of mitochondrial biogenesis, chondrogenic transcription factor SOX9, and cartilage matrix expression of human chondrocytes cultures. Transgenic mice that overexpressed human HSP60 (TgHSP60) had higher chondrocyte proliferation and thicker articular cartilage than wild-type mice. In collagenase-induced OA knees, analyses of CatWalk, 2-deoxyglucose-probed fluorescence imaging, and μCT revealed that affected knees of TgHSP60 mice showed minor footprint irregularity, joint inflammation, and osteophyte formation. HSP60 overexpression also alleviated the histopathology of cartilage damage, synovial hypervascularization, and macrophage infiltration within joint lesions. Intra-articular administration of exogenous HSP60 ameliorated the pathogenesis of cartilage deterioration, synovitis, and osteophyte accumulation, thereby improving gait profiles of the collagenase-injured knees. HSP60 signaling maintains SOX9 levels by attenuating SOX9 hyper-ubiquitination of affected joints. Taken together, HSP60 deficiency in articular compartments was relevant to OA knee incidence. Sustained HSP60 signaling is favorable to mitigate the progression of OA. This study highlights the joint-anabolic actions of HSP60 and provides perspective on its therapeutic potential for OA.

Key messages

  • HSP60 deficiency is relevant to the existence of end-stage knee osteoarthritis.

  • HSP60 overexpression attenuates cartilage matrix loss of inflammatory chondrocytes.

  • HSP60 transgenic mice showed mild articular injury during knee osteoarthritis.

  • HSP60 maintains knee joint homeostasis through reducing SOX9 ubiquitination.

  • Control of cartilage-anabolic regulator HSP60 ameliorates knee osteoarthritis.

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References

  1. Felson D, Niu J, Sack B, Aliabadi P, McCullough C, Nevitt MC (2013) Progression of osteoarthritis as a state of inertia. Ann Rheum Dis 72:924–929

    Article  PubMed  Google Scholar 

  2. Loeser RF, Goldring SR, Scanzello CR, Goldring MB (2012) Osteoarthritis: a disease of the joint as organ. Arthritis Rheum 64:1697–1707

    Article  PubMed  PubMed Central  Google Scholar 

  3. de Lange-Brokaar BJ, Ioan-Facsinay A, van Osch GJ, Zuurmond AM, Schoones J, Toes RE, Huizinga TW, Kloppenburg M (2012) Synovial inflammation, immune cells and their cytokines in osteoarthritis: a review. Osteoarthr Cartil 20:1484–1499

    Article  PubMed  Google Scholar 

  4. O’Conor CJ, Leddy HA, Benefield HC, Liedtke WB, Guilak F (2014) TRPV4-mediated mechanotransduction regulates the metabolic response of chondrocytes to dynamic loading. Proc Natl Acad Sci U S A 111:1316–1321

    Article  PubMed  PubMed Central  Google Scholar 

  5. Culley KL, Hui W, Barter MJ, Davidson RK, Swingler TE, Destrument AP, Scott ST, Dowell ST, Fenwick S, Rowan AD et al (2013) Class I histone deacetylase inhibition modulates metalloproteinase expression and blocks cytokine-induced cartilage degradation. Arthritis Rheum 65:1822–1830

    Article  CAS  PubMed  Google Scholar 

  6. Bastinaansen-Jenniskens YM, Wei W, Feijt C, Waarsing JH, Verhaar JA, Zuurmond AM, Hanemaaijer R, Stoop R, van Osch GJ (2013) Stimulation of fibrotic processes by the infrapatellar fat pad in cultured synoviocytes from patients with osteoarthritis: a possible role for prostaglandin f2α. Arthritis Rheum 65:2070–2080

    Article  Google Scholar 

  7. Weng T, Yi L, Huang J, Luo F, Wen X, Du X, Chen Q, Deng C, Chen L (2012) Genetic inhibition of fibroblast growth factor receptor 1 in knee cartilage attenuates the degeneration of articular cartilage in adult mice. Arthritis Rheum 64:3982–3992

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Arlet JB, Ribeil JA, Guillem F, Negre O, Hazoume A, Marcion G, Beuzard Y, Dussiot M, Moura IC, Demarest S et al (2014) HSP70 sequestration by α-globin promotes ineffective erythropoiesis in β-thalassaemia. Nature 514:242–246

    CAS  PubMed  Google Scholar 

  9. Winter L, Staszewska I, Mihailovska E, Fischer I, Goldmann WH, Schröder R, Wiche G (2014) Chemical chaperone ameliorates pathological protein aggregation in plectin-deficient muscle. J Clin Invest 124:1144–1157

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Ghosh A, Stuehr DJ (2012) Soluble guanylyl cyclase requires heat shock protein 90 for heme insertion during maturation of the NO-active enzyme. Proc Natl Acad Sci U S A 109:12998–13003

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Koya T, Nishizawa S, Ohno Y, Goto A, Ikuta A, Suzuki M, Ohira T, Egawa T, Nakai A, Sugiura T et al (2013) Heat shock transcription factor-1 deficiency attenuates overloading-associated hypertrophy of mouse soleus muscle. PLoS One 8, e77788

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. van Herwijnen MJ, Wieten L, van der Zee R, van Kooten PJ, Wagenaar-Hilbers JP, Hoek A, den Braber I, Anderton SM, Singh M, Meiring HD et al (2012) Regulatory T cells that recognize a ubiquitous stress-induced self-antigen are long-lived suppressors of autoimmune arthritis. Proc Natl Acad Sci U S A 109:14134–14139

    Article  PubMed  PubMed Central  Google Scholar 

  13. Siebelt M, Jahr H, Groen HC, Sandker M, Waarsing JH, Kops N, Müller C, van Eden W, de Jong M, Weinans H (2013) Hsp90 inhibition protects against biomechanically induced osteoarthritis. Arthritis Rheum 65:2102–2112

    Article  CAS  PubMed  Google Scholar 

  14. Yoo SA, You D, Yoon HJ, Kim DH, Kim HS, Lee K, Ahn JH, Hwang D, Lee AS, Kim KJ et al (2012) A novel pathogenic role of the ER chaperone GRP78/BiP in rheumatoid arthritis. J Exp Med 209:871–886

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Tsuchida S, Arai Y, Takahashi KA, Kishida T, Terauchi R, Honjo K, Nakagawa S, Inoue H, Ikoma K, Ueshima K et al (2014) HIF-1α-induced HSP70 regulates anabolic responses in articular chondrocytes under hypoxic conditions. J Orthop Res 32:975–980

    Article  CAS  PubMed  Google Scholar 

  16. Lambrecht S, Dhaenens M, Almqvist F, Verdonk P, Verbruggen G, Deforce D, Elewaut D (2010) Proteome characterization of human articular chondrocytes leads to novel insights in the function of small heat-shock protein in chondrocyte homeostasis. Osteoarthr Cartil 18:440–446

    Article  CAS  PubMed  Google Scholar 

  17. Fan Z, Tardif G, Hum D, Duval N, Pelletier JP, Martel-Pelletier J (2009) HSP90(beta) and p130(cas): novel regulatory factors of MMP-13 expression in human osteoarthritis chondrocytes. Ann Rheum Dis 68:976–982

    Article  CAS  PubMed  Google Scholar 

  18. Christensen JH, Nielsen MN, Hansen J, Füchtbauer A, Füchtbauer EM, West M, Corydon TJ, Gregersen N, Bross P (2010) Inactivation of the hereditary spastic paraplegia-associated Hspd1 gene encoding the Hsp60 chaperone results in early embryonic lethality in mice. Cell Stress Chaperones 15:851–863

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Kleninridders A, Lauritzen HP, Ussar S, Christensen JH, Mori MA, Bross P, Kahn CR (2013) Leptin regulation of HSP60 impacts hypothalamic insulin signaling. J Clin Invest 123:4667–4680

    Article  Google Scholar 

  20. Suwanwela J, Farber CR, Haung BL, Song B, Pan C, Lyons KM, Lusis AJ (2011) Systems genetics analysis of mouse chondrocyte differentiation. J Bone Miner Res 26:747–760

    Article  CAS  PubMed  Google Scholar 

  21. Romanello M, Piatkowska E, Antoniali G, Cesaratto L, Vascotto C, Iozzo RV, Delneri D, Brancia FL (2014) Osteoblastic cell secretome: a novel role for progranulin during risedronate treatment. Bone 58:81–91

    Article  CAS  PubMed  Google Scholar 

  22. Zonneveld-Huijssoon E, van Wijk F, Roord S, Delemarre E, Meerding J, de Jager W, Klein M, Raz R, Albani S, Kuis W et al (2012) TLR9 agonist CpG enhances protective nasal HSP60 peptide vaccine efficacy in experimental autoimmune arthritis. Ann Rheum Dis 71:1706–1715

    Article  CAS  PubMed  Google Scholar 

  23. Wang FS, Wu RW, Yeh DW, Chen MW, Ke HC, Wu SL, Ko JY (2011) Heat shock protein 60 protects skeletal tissue against glucocorticoid-induced bone mass loss by regulating osteoblast survival. Bone 49:1080–1089

    Article  CAS  PubMed  Google Scholar 

  24. Weng LH, Ko JY, Wang CJ, Sun YC, Wang FS (2012) Dkk-1 promotes angiogenic responses and cartilage matrix proteinase secretion in synovial fibroblasts from osteoarthritic joints. Arthritis Rheum 64:3267–3277

    Article  CAS  PubMed  Google Scholar 

  25. Hsiao YC, Chang HH, Tsai CY, Jong TJ, Horng LS, Lin SF, Tsai TF (2004) Coat color-tagged green mouse with EGFP expressed from the RNA polymerase II promoter. Genesis 39:122–129

    Article  CAS  PubMed  Google Scholar 

  26. Daans M, Luyten FP, Lories RJ (2011) GDF5 deficiency in mice is associated with instability-driven joint damage, gait and subchondral bone changes. Ann Rheum Dis 70:208–213

    Article  PubMed  Google Scholar 

  27. Licha K, Welker P, Weinhart M, Wegner N, Kern S, Reichert S, Gemeinhardt I, Weissbach C, Ebert B, Haag R et al (2011) Fluorescence imaging with multifunctional polyglycerol sulfates: novel polymeric near-IR probes targeting inflammation. Bioconjug Chem 22:2453–2460

    Article  CAS  PubMed  Google Scholar 

  28. Chang CL, Hsu YT, Wu CC, Yang YC, Wang C, Wu TC, Hung CF (2012) Immune mechanism of the anittumor effects generated by bortezomib. J Immunol 189:3209–3220

    Article  CAS  PubMed  Google Scholar 

  29. Wick G, Jakic B, Buszko M, Wick MC, Grundtman C (2014) The role of heat shock proteins in atherosclerosis. Nat Rev Cardiol 11:516–529

    Article  CAS  PubMed  Google Scholar 

  30. Lambrecht S, Verbruggen G, Elewaut D, Deforce D (2009) Differential expression of alphaB-crystallin and evidence of its role as a mediator of matrix gene expression in osteoarthritis. Arthritis Rheum 60:179–188

    Article  CAS  PubMed  Google Scholar 

  31. Fujita S, Arai Y, Nakagawa S, Takahashi KA, Terauchi R, Inoue A, Tonomura H, Hiraoka M, Inoue H, Tsuchida S et al (2012) Combined microwave irradiation and intraarticular glutamine administration-induced HSP70 expression therapy prevents cartilage degradation in a rat osteoathritis model. J Orthop Res 30:401–407

    Article  CAS  PubMed  Google Scholar 

  32. Bradley EW, Carpio LR, McGee-Lawrence ME, Castillejo Becerra C, Amanatulla DF, Ta LE, Otero M, Goldring MB, Kakar S, Westendorf JJ (2015) Phlpp1 facilitates post-traumatic osteoarthritis and is induced by inflammation and promoter demthylation in human osteoarthritis. Osteoarthr Cartil. doi:10.1016/j.joca.2015.12.014

    PubMed  Google Scholar 

  33. Rushton MD, Reynard LN, Barter MJ, Refaie R, Rankin KS, Young DA, Loughlin J (2014) Characterization of the cartilage DNA methylation in knee and hip osteoarthritis. Arthritis Rheumatol 66:2450–2460

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Wang Y, Zhao X, Lotz M, Terkeltaub R, Liu-Bryan R (2015) Mitochondrial biogenesis is impaired in osteoarthritis chondrocytes but reversible via peroxisome proliferator-activated receptor γ coactivator 1α. Arthritis Rheum 67:2141–2153

    Article  CAS  Google Scholar 

  35. den Hollanders W, Ramos YF, Bos SD, Bomer N, van der Breggen R, Lakenberg N, de Dijcker WJ, Duijnisveld BJ, Slagboom PE, Nelissen RG et al (2014) Knee and hip articular cartilage have distinct epigenomic landscapes: implications for future cartilage regeneration approaches. Ann Rheum Dis 73:2208–2212

    Article  Google Scholar 

  36. Kapoor M, Martel-Pelletier J, Lajeunesse D, Pelletier JP, Fahmi H (2011) Role of proinflammatiory cytokines in the pathophysiology of osteoarthritis. Nat Rev Rheumatol 7:33–42

    Article  CAS  PubMed  Google Scholar 

  37. Leavenworth JW, Tang X, Kim HJ, Wang X, Cantor H (2013) Amelioration of arthritis through mobilization of peptide-specific CD8+ regulatory T cells. J Clin Invest 123:1382–1389

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Liu SC, Chuang SM, Hsu CJ, Tsai CH, Wang SW, Tang CH (2014) CTGF increases vascular endothelial growth factor-dependent angiogenesis in human synovial fibroblasts by increasing miR-210 expression. Cell Death Dis 5, e1485

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Koelling S, Kruegel J, Irmer M, Path JR, Sadowski B, Miro X, Miosge N (2009) Migratory chondrogenic progenitor cells from repair tissue during the later stages of human osteoarthritis. Cell Stem Cell 4:324–335

    Article  CAS  PubMed  Google Scholar 

  40. Radwan M, Wilkinson DJ, Hui W, Destrument A, Charlton SH, Barter MJ, Gibson B, Coulombe J, Gray DA, Rowan AD et al (2015) Protection against murine osteoarthritis by inhibition of the 26 proteasome and lysine-48 linked ubiquitination. Ann Rheum Dis 74:1580–1587

    Article  CAS  PubMed  Google Scholar 

  41. Hattori T, Kishino T, Stephen S, Eberspaecher H, Maki S, Takigawa M, de Crombrugghe B, Yasuda H (2013) E6-AP/UBE3A protein acts as a ubiquitin ligase toward SOX9 protein. J Biol Chem 288:35138–35148

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Tsai YP, Yang MH, Huang CH, Chang SY, Chen PM, Liu CJ, Teng SC, Wu KJ (2009) Interaction between HSP60 and beta-catenin promotes metastasis. Carcinogenesis 30:1049–1057

    Article  CAS  PubMed  Google Scholar 

  43. Yang Y, Fiskus W, Yang B, Atadja P, Takahashi Y, Pandita TK, Wang HG, Bhalla KN (2013) Acetylated hsp70 and KAP1-mediated Vps34 SUMOylation is required for autophagosome creation in autophagy. Proc Natl Acad Sci U S A 110:6841–6846

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Li Y, Wang Y, Chubinskaya S, Schoeberl B, Florine E, Kopesky P, Grodzinsky AJ (2015) Effects of insulin-like growth factor-1 and dexamethasone on cytokine-challenged cartilage: relevance to post-traumatic osteoarthritis. Osteoarthr Cartil 23:266–274

    Article  CAS  PubMed  Google Scholar 

  45. Loffredo FS, Pancoast JR, Cai L, Vannelli T, Dong JZ, Lee RT, Patwari P (2014) Targeted delivery to cartilage is critical for in vivo efficacy of insulin-like growth factor in a rat model of osteoarthritis. Arthritis Rheumatol 66:1247–1255

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study was partially supported by grants [NSC101-2314-B-182A-117] and [NSC100-2314-B-182A-028-MY3] from the Ministry of Science & Technology, [NHRI-EX104-10536SI] from the National Health Research Institute and [CMRPG8A0423, CMRPG8B0873, CMRPG8B0883, and CLRPG8B0042] from Chang Gung Memorial Hospital, Taiwan. We are grateful to the Center for Laboratory Animals, Kaohisung Chang Gung Memorial Hospital, Taiwan, for use of their facilities.

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Authors and Affiliations

  1. Department of Orthopedic Surgery, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan

    Jih-Yang Ko

  2. Core Laboratory for Phenomics and Diagnostics, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan

    Yi-Chih Sun, Wen-Chin Li & Feng-Sheng Wang

  3. Graduate Institute of Clinical Medical Sciences, Chang Gung University College of Medicine, Kaohsiung, Taiwan

    Yi-Chih Sun, Wen-Chin Li & Feng-Sheng Wang

  4. Department of Medical Research, Kaohsiung Chang Gung Memorial Hospital, 123 Ta-Pei Road, Niao Sung District, Kaohsiung, 833, Taiwan

    Yi-Chih Sun, Wen-Chin Li & Feng-Sheng Wang

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  1. Jih-Yang Ko
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Correspondence to Feng-Sheng Wang.

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This study was approved by the IRB (No.101-2684B) of Chang Gung Memorial Hospital. Informed consent was obtained from all participants. Animal use protocols were approved by the IACUC of Chang Gung Memorial Hospital (No.2011121403).

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Jih-Yang Ko, Yi-Chih Sun, Wen-Chin Li, and Feng-Sheng Wang have nothing to disclose.

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J.-Y. Ko and Y.-C. Sun contributed equally to this work.

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Ko, JY., Sun, YC., Li, WC. et al. Chaperonin 60 regulation of SOX9 ubiquitination mitigates the development of knee osteoarthritis. J Mol Med 94, 755–769 (2016). https://doi.org/10.1007/s00109-016-1422-3

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  • DOI: https://doi.org/10.1007/s00109-016-1422-3

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