Histone demethylase JMJD3 downregulation protects against aberrant force-induced osteoarthritis through epigenetic control of NR4A1

Abstract

Osteoarthritis (OA) is a prevalent joint disease with no effective treatment strategies. Aberrant mechanical stimuli was demonstrated to be an essential factor for OA pathogenesis. Although multiple studies have detected potential regulatory mechanisms underlying OA and have concentrated on developing novel treatment strategies, the epigenetic control of OA remains unclear. Histone demethylase JMJD3 has been reported to mediate multiple physiological and pathological processes, including cell differentiation, proliferation, autophagy, and apoptosis. However, the regulation of JMJD3 in aberrant force-related OA and its mediatory effect on disease progression are still unknown. In this work, we confirmed the upregulation of JMJD3 in aberrant force-induced cartilage injury in vitro and in vivo. Functionally, inhibition of JMJD3 by its inhibitor, GSK-J4, or downregulation of JMJD3 by adenovirus infection of sh-JMJD3 could alleviate the aberrant force-induced chondrocyte injury. Mechanistic investigation illustrated that aberrant force induces JMJD3 expression and then demethylates H3K27me3 at the NR4A1 promoter to promote its expression. Further experiments indicated that NR4A1 can regulate chondrocyte apoptosis, cartilage degeneration, extracellular matrix degradation, and inflammatory responses. In vivo, anterior cruciate ligament transection (ACLT) was performed to construct an OA model, and the therapeutic effect of GSK-J4 was validated. More importantly, we adopted a peptide-siRNA nanoplatform to deliver si-JMJD3 into articular cartilage, and the severity of joint degeneration was remarkably mitigated. Taken together, our findings demonstrated that JMJD3 is flow-responsive and epigenetically regulates OA progression. Our work provides evidences for JMJD3 inhibition as an innovative epigenetic therapy approach for joint diseases by utilizing p5RHH-siRNA nanocomplexes.

References

[1] Lu KH, . The potential remedy of melatonin on osteoarthritis. J. Pineal Res., 2021, 71: e12762.

[2] Li X, . Nanoparticle-Cartilage Interaction: Pathology-Based Intra-articular Drug Delivery for Osteoarthritis Therapy. Nanomicro Lett., 2021, 13: 149.

[3] Yokota H, Goldring MB, Sun HB. CITED2-mediated regulation of MMP-1 and MMP-13 in human chondrocytes under flow shear. J. Biol. Chem., 2003, 278: 47275-47280.

[4] Buckwalter JA, Martin JA, Brown TD. Perspectives on chondrocyte mechanobiology and osteoarthritis. Biorheology, 2006, 43: 603-609.

[5] Kerin A, Patwari P, Kuettner K, Cole A, Grodzinsky A. Molecular basis of osteoarthritis: biomechanical aspects. Cell Mol. Life Sci., 2002, 59: 27-35.

[6] Carter DR, Wong M. Modelling cartilage mechanobiology. Philos. Trans. R. Soc. Lond. B Biol. Sci., 2003, 358: 1461-1471.

[7] Carter DR, . The mechanobiology of articular cartilage development and degeneration. Clin. Orthop. Relat. Res., 2004, 427S: S69-S77.

[8] Donahue TL, Fisher MB, Maher SA. Meniscus mechanics and mechanobiology. J. Biomech., 2015, 48: 1341-1342.

[9] Guilak F. Biomechanical factors in osteoarthritis. Best. Pr. Res. Clin. Rheumatol., 2011, 25: 815-823.

[10] Rice SJ, Beier F, Young DA, Loughlin J. Interplay between genetics and epigenetics in osteoarthritis. Nat. Rev. Rheumatol., 2020, 16: 268-281.

[11] Allis CD, Jenuwein T. The molecular hallmarks of epigenetic control. Nat. Rev. Genet., 2016, 17: 487-500.

[12] Lorzadeh A, Romero-Wolf M, Goel A, Jadhav U. Epigenetic Regulation of Intestinal Stem Cells and Disease: A Balancing Act of DNA and Histone Methylation. Gastroenterology, 2021, 160: 2267-2282.

[13] Greer EL, Shi Y. Histone methylation: a dynamic mark in health, disease and inheritance. Nat. Rev. Genet., 2012, 13: 343-357.

[14] Wei, Y. et al. Efficient derivation of human trophoblast stem cells from primed pluripotent stem cells. Sci. Adv. 7, https://doi.org/10.1126/sciadv.abf4416 (2021).

[15] Wu C, . Interplay of m(6)A and H3K27 trimethylation restrains inflammation during bacterial infection. Sci. Adv., 2020, 6: eaba0647.

[16] Suwala AK, . Oligosarcomas, IDH-mutant are distinct and aggressive. Acta Neuropathol., 2022, 143: 263-281.

[17] Xiang Y, . JMJD3 is a histone H3K27 demethylase. Cell Res., 2007, 17: 850-857.

[18] Nguyen K, . Inhibition of the H3K27 demethylase UTX enhances the epigenetic silencing of HIV proviruses and induces HIV-1 DNA hypermethylation but fails to permanently block HIV reactivation. PLoS Pathog., 2021, 17: e1010014.

[19] Sun D, Cao X, Wang C. Polycomb chromobox Cbx2 enhances antiviral innate immunity by promoting Jmjd3-mediated demethylation of H3K27 at the Ifnb promoter. Protein Cell, 2019, 10: 285-294.

[20] Ding Y, . JMJD3: a critical epigenetic regulator in stem cell fate. Cell Commun. Signal, 2021, 19

[21] Lee SH, Kim O, Kim HJ, Hwangbo C, Lee JH. Epigenetic regulation of TGF-beta-induced EMT by JMJD3/KDM6B histone H3K27 demethylase. Oncogenesis, 2021, 10

[22] Salminen A, Kaarniranta K, Hiltunen M, Kauppinen A. Histone demethylase Jumonji D3 (JMJD3/KDM6B) at the nexus of epigenetic regulation of inflammation and the aging process. J. Mol. Med (Berl.), 2014, 92: 1035-1043.

[23] Merkwirth C, . Two Conserved Histone Demethylases Regulate Mitochondrial Stress-Induced Longevity. Cell, 2016, 165: 1209-1223.

[24] Liao, Y. et al. Inhibition of EZH2 transactivation function sensitizes solid tumors to genotoxic stress. Proc. Natl Acad. Sci. USA 119, https://doi.org/10.1073/pnas.2105898119 (2022).

[25] Krauss, L. et al. HDAC2 facilitates pancreatic cancer metastasis. Cancer Res. https://doi.org/10.1158/0008-5472.CAN-20-3209 (2021).

[26] Connor MG, . The histone demethylase KDM6B fine-tunes the host response to Streptococcus pneumoniae. Nat. Microbiol., 2021, 6: 257-269.

[27] Wu W, . Cystathionine-gamma-lyase ameliorates the histone demethylase JMJD3-mediated autoimmune response in rheumatoid arthritis. Cell Mol. Immunol., 2019, 16: 694-705.

[28] Barrett T, . NCBI GEO: archive for functional genomics data sets-update. Nucleic Acids Res., 2013, 41: D991-D995.

[29] Kruidenier L, . A selective jumonji H3K27 demethylase inhibitor modulates the proinflammatory macrophage response. Nature, 2012, 488: 404-408.

[30] Zhang X, Liu L, Yuan X, Wei Y, Wei X. JMJD3 in the regulation of human diseases. Protein Cell, 2019, 10: 864-882.

[31] Saito T, . Transcriptional regulation of endochondral ossification by HIF-2alpha during skeletal growth and osteoarthritis development. Nat. Med., 2010, 16: 678-686.

[32] Yang S, . Hypoxia-inducible factor-2alpha is a catabolic regulator of osteoarthritic cartilage destruction. Nat. Med., 2010, 16: 687-693.

[33] Li ZC, . Functional annotation of rheumatoid arthritis and osteoarthritis associated genes by integrative genome-wide gene expression profiling analysis. PLoS ONE, 2014, 9: e85784.

[34] Wang S, . HUCMSCs transplantation combined with ultrashort wave therapy attenuates neuroinflammation in spinal cord injury through NUR77/ NF-kappaB pathway. Life Sci., 2021, 267: 118958.

[35] Cao J, . NR4A1 knockdown confers hepatoprotection against ischaemia-reperfusion injury by suppressing TGFbeta1 via inhibition of CYR61/NF-kappaB in mouse hepatocytes. J. Cell Mol. Med, 2021, 25: 5099-5112.

[36] Shi Z, . Hypoxia-induced Nur77 activates PI3K/Akt signaling via suppression of Dicer/let-7i-5p to induce epithelial-to-mesenchymal transition. Theranostics, 2021, 11: 3376-3391.

[37] Wight L, Owen D, Goldbloom D, Knupp M. Pure Ankle Dislocation: a systematic review of the literature and estimation of incidence. Injury, 2017, 48: 2027-2034.

[38] Jiang Y, . KDM6B-mediated histone demethylation of LDHA promotes lung metastasis of osteosarcoma. Theranostics, 2021, 11: 3868-3881.

[39] Mallaney C, . Kdm6b regulates context-dependent hematopoietic stem cell self-renewal and leukemogenesis. Leukemia, 2019, 33: 2506-2521.

[40] Li J, . KDM6B-dependent chromatin remodeling underpins effective virus-specific CD8(+) T cell differentiation. Cell Rep., 2021, 34: 108839.

[41] Chang L, . H3K27 demethylase KDM6B aggravates ischemic brain injury through demethylation of IRF4 and Notch2-dependent SOX9 activation. Mol. Ther. Nucleic Acids, 2021, 24: 622-633.

[42] Maleszewska M, Vanchin B, Harmsen MC, Krenning G. The decrease in histone methyltransferase EZH2 in response to fluid shear stress alters endothelial gene expression and promotes quiescence. Angiogenesis, 2016, 19: 9-24.

[43] Zhang Z, . Mechanical force-sensitive lncRNA SNHG8 inhibits osteogenic differentiation by regulating EZH2 in hPDLSCs. Cell Signal, 2022, 93: 110285.

[44] Datta, H. K. et al. Mechanical-Stress-Related Epigenetic Regulation of ZIC1 Transcription Factor in the Etiology of Postmenopausal Osteoporosis. Int. J. Mol. Sci. 23, https://doi.org/10.3390/ijms23062957 (2022).

[45] Zheng T, . Long non-coding RNA HOTAIRincreased mechanical stimulation-induced apoptosis by regulating microRNA-221/BBC3 axis in C28/I2 cells. Bioengineered, 2021, 12: 10734-10744.

[46] Huang Z, . BRD4 inhibition alleviates mechanical stress-induced TMJ OA-like pathological changes and attenuates TREM1-mediated inflammatory response. Clin. Epigenet., 2021, 13

[47] Yasui T, . Epigenetic regulation of osteoclast differentiation: possible involvement of Jmjd3 in the histone demethylation of Nfatc1. J. Bone Min. Res., 2011, 26: 2665-2671.

[48] LaMere SA, . H3K27 Methylation Dynamics during CD4 T Cell Activation: Regulation of JAK/STAT and IL12RB2 Expression by JMJD3. J. Immunol., 2017, 199: 3158-3175.

[49] Li Q, . Critical role of histone demethylase Jmjd3 in the regulation of CD4+ T-cell differentiation. Nat. Commun., 2014, 5

[50] Huang M, . Jmjd3 regulates inflammasome activation and aggravates DSS-induced colitis in mice. FASEB J., 2020, 34: 4107-4119.

[51] Donas C, . The histone demethylase inhibitor GSK-J4 limits inflammation through the induction of a tolerogenic phenotype on DCs. J. Autoimmun., 2016, 75: 105-117.

[52] Hou KK, Pan H, Ratner L, Schlesinger PH, Wickline SA. Mechanisms of nanoparticle-mediated siRNA transfection by melittin-derived peptides. ACS Nano, 2013, 7: 8605-8615.

[53] Zhou HF, . Peptide-siRNA nanocomplexes targeting NF-kappaB subunit p65 suppress nascent experimental arthritis. J. Clin. Investig., 2014, 124: 4363-4374.

[54] Yan H, . Suppression of NF-kappaB activity via nanoparticle-based siRNA delivery alters early cartilage responses to injury. Proc. Natl Acad. Sci. USA, 2016, 113: E6199-E6208.

[55] Winoto A, Littman DR. Nuclear hormone receptors in T lymphocytes. Cell, 2002, 109: S57-S66. Suppl

[56] Lee JM, Lee KH, Weidner M, Osborne BA, Hayward SD. Epstein-Barr virus EBNA2 blocks Nur77- mediated apoptosis. Proc. Natl Acad. Sci. USA, 2002, 99: 11878-11883.

[57] Zhou F, . Nuclear receptor NR4A1 promotes breast cancer invasion and metastasis by activating TGF-beta signalling. Nat. Commun., 2014, 5

[58] Maijenburg MW, van der Schoot CE, Voermans C. Mesenchymal stromal cell migration: possibilities to improve cellular therapy. Stem Cells Dev., 2012, 21: 19-29.

[59] Landry ES, Rouillard C, Levesque D, Guertin PA. Profile of immediate early gene expression in the lumbar spinal cord of low-thoracic paraplegic mice. Behav. Neurosci., 2006, 120: 1384-1388.

[60] Herring, J. A., Elison, W. S. & Tessem, J. S. Function of Nr4a Orphan Nuclear Receptors in Proliferation, Apoptosis and Fuel Utilization Across Tissues. Cells 8, https://doi.org/10.3390/cells8111373 (2019).

[61] Shi X, Ye H, Yao X, Gao Y. The involvement and possible mechanism of NR4A1 in chondrocyte apoptosis during osteoarthritis. Am. J. Transl. Res., 2017, 9: 746-754.

[62] Zheng Z, . NR4A1 promotes TNFalphainduced chondrocyte death and migration injury via activating the AMPK/Drp1/mitochondrial fission pathway. Int J. Mol. Med., 2020, 45: 151-161.

[63] Jin Y, . Aberrant Fluid Shear Stress Contributes to Articular Cartilage Pathogenesis via Epigenetic Regulation of ZBTB20 by H3K4me3. J. Inflamm. Res., 2021, 14: 6067-6083.

[64] Ruan MZ, Patel RM, Dawson BC, Jiang MM, Lee BH. Pain, motor and gait assessment of murine osteoarthritis in a cruciate ligament transection model. Osteoarthr. Cartil., 2013, 21: 1355-1364.

[65] Pritzker KP, . Osteoarthritis cartilage histopathology: grading and staging. Osteoarthr. Cartil., 2006, 14: 13-29.

Funding

National Natural Science Foundation of China (National Science Foundation of China)(11932012, 81870790, 31801233, 31801233)

Science and Technology Commission of Shanghai Municipality (Shanghai Municipal Science and Technology Commission)(18441903600)

Clinical Research Plan of SHDC (No. SHDC2020CR3009A) Innovative Research Team of High-level Local Universities in Shanghai (SSMU-ZDCX20180902)