Matrix stiffening promotes chondrocyte senescence and the osteoarthritis development through downregulating HDAC3

Abstract

Extracellular matrix (ECM) stiffening is a typical characteristic of cartilage aging, which is a quintessential feature of knee osteoarthritis (KOA). However, little is known about how ECM stiffening affects chondrocytes and other molecules downstream. This study mimicked the physiological and pathological stiffness of human cartilage using polydimethylsiloxane (PDMS) substrates. It demonstrated that epigenetic Parkin regulation by histone deacetylase 3 (HDAC3) represents a new mechanosensitive mechanism by which the stiffness matrix affected chondrocyte physiology. We found that ECM stiffening accelerated cultured chondrocyte senescence in vitro, while the stiffness ECM downregulated HDAC3, prompting Parkin acetylation to activate excessive mitophagy and accelerating chondrocyte senescence and osteoarthritis (OA) in mice. Contrarily, intra-articular injection with an HDAC3-expressing adeno-associated virus restored the young phenotype of the aged chondrocytes stimulated by ECM stiffening and alleviated OA in mice. The findings indicated that changes in the mechanical ECM properties initiated pathogenic mechanotransduction signals, promoted the Parkin acetylation and hyperactivated mitophagy, and damaged chondrocyte health. These results may provide new insights into chondrocyte regulation by the mechanical properties of ECM, suggesting that the modification of the physical ECM properties may be a potential OA treatment strategy.

References

1. Martel-Pelletier, J. et al. Osteoarthritis. Nat. Rev. Dis. Prim. 2, 16072(2016).
2. Sharma L.Osteoarthritis of the knee. N. Engl. J. Med. 384, 51-59 (2021).
3. Glyn-Jones, S.et al. Osteoarthritis. Lancet 386, 376-387 (2015).
4. Zhang, H.et al.Maintaining hypoxia environment of subchondral bone alleviates osteoarthritis progression. Sci. Adv. 9, eabo7868 (2023).
5. Loeser, R. F., Collins, J. A.& Diekman, B. O. Ageing and the pathogenesis of osteoarthritis. Nat. Rev. Rheumatol. 12, 412-420 (2016).
6. De Belly, H., Paluch, E. K. & Chalut, K. J. Interplay between mechanics and signalling in regulating cell fate. Nat. Rev. Mol. Cell Biol. 23, 465-480 (2022).
7. Iijima, H.et al.Age-related matrix stiffening epigenetically regulates α-Klotho expression and compromises chondrocyte integrity. Nat. Commun. 14, 18(2023).
8. Armiento, A. R., Alini, M.& Stoddart, M. J. Articular fibrocartilage—why does hyaline cartilage fail to repair? Adv. Drug Deliv. Rev. 146, 289-305 (2019).
9. Jiang, W.et al.Mechanisms linking mitochondrial mechanotransduction and chondrocyte biology in the pathogenesis of osteoarthritis. Ageing Res. Rev. 67, 101315(2021).
10. Richard, D.et al. Evolutionary selection and constraint on human knee chondrocyte regulation impacts osteoarthritis risk. Cell 181, 362-381.e328 (2020).
11. Zhang, H.et al.Mechanical overloading promotes chondrocyte senescence and osteoarthritis development through downregulating FBXW7. Ann. Rheum. Dis. 81, 676-686 (2022).
12. Wang, S.et al.Mechanical overloading induces GPX4-regulated chondrocyte ferroptosis in osteoarthritis via Piezo1 channel-facilitated calcium influx. J. Adv. Res. 41, 63-75 (2022).
13. Han S.Osteoarthritis year in review 2022: biology. Osteoarthr. Cartil. 30, 1575-1582 (2022).
14. Peng, Z.et al.The regulation of cartilage extracellular matrix homeostasis in joint cartilage degeneration and regeneration. Biomaterials 268, 120555 (2021).
15. Kim, J. H.et al. Matrix cross-linking-mediated mechanotransduction promotes posttraumatic osteoarthritis. Proc. Natl. Acad. Sci. USA 112, 9424-9429 (2015).
16. Stolz, M.et al.Early detection of aging cartilage and osteoarthritis in mice and patient samples using atomic force microscopy. Nat. Nanotechnol. 4, 186-192 (2009).
17. Wu, B.et al.Stiff matrix induces exosome secretion to promote tumour growth. Nat. Cell Biol. 25, 415-424 (2023).
18. Patwardhan S., Mahadik P., Shetty O.& Sen, S. ECM stiffness-tuned exosomes drive breast cancer motility through thrombospondin-1. Biomaterials 279, 121185 (2021).
19. Xu, M.et al.Transplanted senescent cells induce an osteoarthritis-like condition in mice. J. Gerontol. Ser. A Biol. Sci. Med. Sci. 72, 780-785 (2017).
20. Xie, J.et al.Cellular senescence in knee osteoarthritis: molecular mechanisms and therapeutic implications. Ageing Res. Rev. 70, 101413(2021).
21. Jeon, O. H.et al.Local clearance of senescent cells attenuates the development of post-traumatic osteoarthritis and creates a pro-regenerative environment. Nat. Med. 23, 775-781 (2017).
22. Hayashi, D.et al.Pre-radiographic osteoarthritic changes are highly prevalent in the medial patella and medial posterior femur in older persons: Framingham OA study. Osteoarthr. Cartil. 22, 76-83 (2014).
23. Engeland K.Cell cycle regulation: p53-p21-RB signaling. Cell Death Differ. 29, 946-960 (2022).
24. Baker, D. J.et al. Clearance of p16Ink4a-positive senescent cells delays ageingassociated disorders. Nature 479, 232-236 (2011).
25. Meng, F.et al. MicroRNA-193b-3p regulates chondrogenesis and chondrocyte metabolism by targeting HDAC3. Theranostics 8, 2862-2883 (2018).
26. Esteban-Martínez, L. & Boya, P. BNIP3L/NIX-dependent mitophagy regulates cell differentiation via metabolic reprogramming. Autophagy 14, 915-917 (2018).
27. Peña-Blanco, A. et al. Drp1 modulates mitochondrial stress responses to mitotic arrest. Cell Death Differ. 27, 2620-2634 (2020).
28. Shaltouki A., Hsieh C. H., Kim M. J.& Wang, X. Alpha-synuclein delays mitophagy and targeting Miro rescues neuron loss in Parkinson's models. Acta Neuropathol. 136, 607-620 (2018).
29. Emmett M. J.& Lazar, M. A. Integrative regulation of physiology by histone deacetylase 3. Nat. Rev. Mol. Cell Biol. 20, 102-115 (2019).
30. Narita, T., Weinert, B. T.& Choudhary, C. Functions and mechanisms of nonhistone protein acetylation. Nat. Rev. Mol. Cell Biol. 20, 156-174 (2019).
31. Sun, X.et al. Histone deacetylase inhibitors inhibit cervical cancer growth through Parkin acetylation-mediated mitophagy. Acta Pharm. Sin. B 12, 838-852 (2022).
32. Yi, B., Xu, Q.& Liu, W. An overview of substrate stiffness guided cellular response and its applications in tissue regeneration. Bioact. Mater. 15, 82-102 (2022).
33. Janmey, P. A., Fletcher, D. A.& Reinhart-King, C. A. Stiffness sensing by cells. Physiol. Rev. 100, 695-724 (2020).
34. Vining K. H.& Mooney, D. J. Mechanical forces direct stem cell behaviour in development and regeneration. Nat. Rev. Mol. cell Biol. 18, 728-742 (2017).
35. Lyu, C.et al.Advanced glycation end-products as mediators of the aberrant crosslinking of extracellular matrix in scarred liver tissue. Nat. Biomed. Eng. 7, 1437-1454 (2023).
36. Zeng, L.et al.HDAC3 is crucial in shear- and VEGF-induced stem cell differentiation toward endothelial cells. J. Cell Biol. 174, 1059-1069 (2006).
37. Wallace D. C.A mitochondrial bioenergetic etiology of disease. J. Clin. Investig. 123, 1405-1412 (2013).
38. Forbes J. M.& Thorburn, D. R. Mitochondrial dysfunction in diabetic kidney disease. Nat. Rev. Nephrol. 14, 291-312 (2018).
39. Sorrentino, V., Menzies, K. J.& Auwerx, J. Repairing mitochondrial dysfunction in disease. Annu. Rev. Pharmacol. Toxicol. 58, 353-389 (2018).
40. Palikaras, K., Lionaki, E.& Tavernarakis, N. Mechanisms of mitophagy in cellular homeostasis, physiology and pathology. Nat. Cell Biol. 20, 1013-1022 (2018).
41. Liu, L.et al.The physiological metabolite α-ketoglutarate ameliorates osteoarthritis by regulating mitophagy and oxidative stress. Redox Biol. 62, 102663(2023).
42. Jin, Z.et al.Curcumin exerts chondroprotective effects against osteoarthritis by promoting AMPK/PINK1/Parkin-mediated mitophagy. Biomed. Pharmacother. 151, 113092(2022).
43. Ansari M. Y., Khan N. M., Ahmad I.& Haqqi, T. M. Parkin clearance of dysfunctional mitochondria regulates ROS levels and increases survival of human chondrocytes. Osteoarthr. Cartil. 26, 1087-1097 (2018).
44. Wang, C.et al. Protective effects of metformin against osteoarthritis through upregulation of SIRT3-mediated PINK1/Parkin-dependent mitophagy in primary chondrocytes. Biosci. Trends 12, 605-612 (2019).
45. Shin, H. J.et al.Pink1-mediated chondrocytic mitophagy contributes to cartilage degeneration in osteoarthritis. J. Clin. Med. 8, 1849(2019).
46. Kuang, Z.et al. The intestinal microbiota programs diurnal rhythms in host metabolism through histone deacetylase 3. Science 365, 1428-1434 (2019).
47. Wang, Z.et al. SETD5-coordinated chromatin reprogramming regulates adaptive resistance to targeted pancreatic cancer therapy. Cancer Cell 37, 834-849.e813 (2020).
48. Petrucelli, L.et al. Parkin protects against the toxicity associated with mutant alpha-synuclein: proteasome dysfunction selectively affects catecholaminergic neurons. Neuron 36, 1007-1019 (2002).
49. Wang, R.et al. PINK1, Keap1,Rtnl1 regulate selective clearance of endoplasmic reticulum during development. Cell 186, 4172-4188.e4118 (2023).
50. Tamaddon, M.et al.Osteochondral scaffolds for early treatment of cartilage defects in osteoarthritic joints: from bench to clinic. Biomater. Transl. 1, 3-17 (2020).
51. Donate, R.et al.Translation through collaboration: practice applied in BAMOS project in in vivo testing of innovative osteochondral scaffolds. Biomater. Transl. 3, 102-104 (2022).
52. Wang, Y., Chen, Y.& Wei, Y. Osteoarthritis animal models for biomaterial-assisted osteochondral regeneration. Biomater. Transl. 3, 264-279 (2022).
53. Perez-Riverol, Y. et al. The PRIDE database resources in 2022: a hub for mass spectrometry-based proteomics evidences. Nucleic Acids Res. 50, D543-d552 (2022).

Funding

Huan Liu ([e-mail]) or Wenhua Huang ([e-mail])