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Regulation of circADAMTS6-miR-324-5p-PIK3R3 ceRNA pathway may be a novel mechanism of IL-1β-induced osteoarthritic chondrocytes

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Journal of Bone and Mineral Metabolism Aims and scope Submit manuscript

Abstract

Introduction

A disintegrin and metallopeptidase with thrombospondin type 1 motif 6 (ADAMTS6)-derived circular RNA (circADAMTS6; hsa_circ_0008667) is a novel regulator in interleukin (IL)-1β-induced apoptosis of human chondrocytes (HCs). Here, we planned to probe into its role and mechanism underlying HCs injury in osteoarthritis.

Materials and Methods

Real time-quantitative PCR and immunoblotting estimated the abundance of RNA and protein, respectively. Cell proliferation and apoptosis were measured by WST-8, EdU, fluorescein isothiocyanate, and caspase3/7 activity assays. Levels of inflammatory cytokines (IL-6 and tumor necrosis factor-α), apoptosis-related proteins (Bcl-2 and Bcl-2-associated X protein), extracellular matrix (ECM)-related proteins (matrix metalloproteinase-13 and collagen type II alpha-1), and PI3K/AKT/mTOR signaling pathway-related proteins (AKT, mTOR, phosphorylated-AKT, and phosphorylated-mTOR) were evaluated by enzyme-linked immunosorbent assays and immunoblotting. Target relationship was confirmed by dual-luciferase reporter, Argonaute-2 immunoprecipitation and RNA pull-down assays.

Results

Abundances of circADAMTS6 and phosphoinositide-3-kinase regulatory subunit 3 (PIK3R3) were underexpressed, and microRNA (miR)-324-5p was elevated in human osteoarthritic tissues and IL-1β-induced HCs. Overexpressing circADAMTS6 and inhibiting miR-324-5p enhanced proliferation and ECM synthesis, but suppressed apoptosis and inflammatory response in IL-1β-challenged HCs. Besides, silencing circADAMTS6 caused similar effects of IL-1β stress on HCs. Mechanically, there was a direct interaction between miR-324-5p and circADAMTS6 or PIK3R3, and IL-1β-induced activation of PI3K/AKT/mTOR signaling pathway was suppressed by circADAMTS6 overexpression and miR-324-5p silencing. Furthermore, counteractive effects of miR-324-5p upregulation on circADAMTS6 overexpression and PIK3R3 knockdown on miR-324-5p silencing were observed.

Conclusion

CircADAMTS6-miR-324-5p-PIK3R3 axis might participate in IL-1β-induced HCs dysfunction via competing endogenous RNA mechanism and the PI3K/AKT/mTOR signaling pathway.

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References

  1. Xie F, Kovic B, Jin X, He X, Wang M, Silvestre C (2016) Economic and humanistic burden of osteoarthritis: a systematic review of large sample studies. Pharmacoeconomics 34:1087–1100

    Article  Google Scholar 

  2. Global Burden of Disease Study C (2015) Global, regional, and national incidence, prevalence, and years lived with disability for 301 acute and chronic diseases and injuries in 188 countries, 1990–2013: a systematic analysis for the Global Burden of Disease Study 2013. Lancet 386:743–800

    Article  Google Scholar 

  3. Chou CH, Jain V, Gibson J, Attarian DE, Haraden CA, Yohn CB, Laberge RM, Gregory S, Kraus VB (2020) Synovial cell cross-talk with cartilage plays a major role in the pathogenesis of osteoarthritis. Sci Rep 10:10868

    Article  CAS  Google Scholar 

  4. Andia I, Maffulli N (2013) Platelet-rich plasma for managing pain and inflammation in osteoarthritis. Nat Rev Rheumatol 9:721–730

    Article  CAS  Google Scholar 

  5. Mead TJ, Apte SS (2018) ADAMTS proteins in human disorders. Matrix Biol 71–72:225–239

    Article  Google Scholar 

  6. Yang CY, Chanalaris A, Troeberg L (2017) ADAMTS and ADAM metalloproteinases in osteoarthritis—looking beyond the “usual suspects.” Osteoarthritis Cartilage 25:1000–1009

    Article  CAS  Google Scholar 

  7. Malemud CJ (2019) Inhibition of MMPs and ADAM/ADAMTS. Biochem Pharmacol 165:33–40

    Article  CAS  Google Scholar 

  8. Larkin J, Lohr TA, Elefante L, Shearin J, Matico R et al (2015) Translational development of an ADAMTS-5 antibody for osteoarthritis disease modification. Osteoarthritis Cartilage 23:1254–1266

    Article  CAS  Google Scholar 

  9. Fu Q, Li L, Wang B, Wu J, Li H, Han Y, Xiang D, Chen Y, Zhu J (2021) CircADAMTS6/miR-431–5p axis regulate interleukin-1beta induced chondrocyte apoptosis. J Gene Med 23:e3304

    CAS  PubMed  Google Scholar 

  10. Greene J, Baird AM, Brady L, Lim M, Gray SG, McDermott R, Finn SP (2017) Circular RNAs: biogenesis, function and role in human diseases. Front Mol Biosci 4:38

    Article  Google Scholar 

  11. Mao X, Cao Y, Guo Z, Wang L, Xiang C (2021) Biological roles and therapeutic potential of circular RNAs in osteoarthritis. Mol Ther Nucleic Acids 24:856–867

    Article  CAS  Google Scholar 

  12. Jiang S, Liu Y, Xu B, Zhang Y, Yang M (2020) Noncoding RNAs: New regulatory code in chondrocyte apoptosis and autophagy. Wiley Interdiscip Rev RNA 11:e1584

    Article  CAS  Google Scholar 

  13. Zhang W, Qi L, Chen R, He J, Liu Z, Wang W, Tu C, Li Z (2021) Circular RNAs in osteoarthritis: indispensable regulators and novel strategies in clinical implications. Arthritis Res Ther 23:23

    Article  CAS  Google Scholar 

  14. Liu D, Liang YH, Yang YT, He M, Cai ZJ, Xiao WF, Li YS (2021) Circular RNA in osteoarthritis: an updated insight into the pathophysiology and therapeutics. Am J Transl Res 13:11–23

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Kulcheski FR, Christoff AP, Margis R (2016) Circular RNAs are miRNA sponges and can be used as a new class of biomarker. J Biotechnol 238:42–51

    Article  CAS  Google Scholar 

  16. Xiao K, Xia Z, Feng B, Bian Y, Fan Y, Li Z, Wu Z, Qiu G, Weng X (2019) Circular RNA expression profile of knee condyle in osteoarthritis by illumina HiSeq platform. J Cell Biochem 120:17500–17511

    Article  CAS  Google Scholar 

  17. Woods S, Barter MJ, Elliott HR, McGillivray CM, Birch MA, Clark IM, Young DA (2019) miR-324-5p is up regulated in end-stage osteoarthritis and regulates Indian Hedgehog signalling by differing mechanisms in human and mouse. Matrix Biol 77:87–100

    Article  CAS  Google Scholar 

  18. Wang WT, Huang ZP, Sui S, Liu JH, Yu DM, Wang WB (2020) microRNA-1236 promotes chondrocyte apoptosis in osteoarthritis via direct suppression of PIK3R3. Life Sci 253:117694

    Article  CAS  Google Scholar 

  19. Murphy WA Jr, Altman RD (1995) Updated osteoarthritis reference standard. J Rheumatol Suppl 43:56–59

    PubMed  Google Scholar 

  20. Guo Y, Min Z, Jiang C, Wang W, Yan J et al (2018) Downregulation of HS6ST2 by miR-23b-3p enhances matrix degradation through p38 MAPK pathway in osteoarthritis. Cell Death Dis 9:699

    Article  Google Scholar 

  21. Sun K, Luo J, Guo J, Yao X, Jing X, Guo F (2020) The PI3K/AKT/mTOR signaling pathway in osteoarthritis: a narrative review. Osteoarthritis Cartilage 28:400–409

    Article  CAS  Google Scholar 

  22. Wan Y, Luo H, Yang M, Tian X, Peng B et al (2020) miR-324-5p contributes to cell proliferation and apoptosis in pancreatic cancer by targeting KLF3. Mol Ther Oncolytics 18:432–442

    Article  CAS  Google Scholar 

  23. Gu C, Zhang M, Sun W, Dong C (2019) Upregulation of miR-324-5p Inhibits proliferation and invasion of colorectal cancer cells by targeting ELAVL1. Oncol Res 27:515–524

    Article  Google Scholar 

  24. Zhou X, Shi X, Wang J, Zhang X, Xu Y, Liu Y, Li X, Yang G (2020) miR-324-5p promotes adipocyte differentiation and lipid droplet accumulation by targeting Krueppel-like factor 3 (KLF3). J Cell Physiol 235:7484–7495

    Article  CAS  Google Scholar 

  25. Guo J, Yang C, Lin Y, Hu G, Wei J, Zhang X, Chen X, Li J (2020) Enhanced peripheral blood miR-324–5p is associated with the risk of metabolic syndrome by suppressing ROCK1. Biochim Biophys Acta Mol Cell Biol Lipids 1865:158727

    Article  CAS  Google Scholar 

  26. Lin AC, Seeto BL, Bartoszko JM, Khoury MA, Whetstone H, Ho L, Hsu C, Ali SA, Alman BA (2009) Modulating hedgehog signaling can attenuate the severity of osteoarthritis. Nat Med 15:1421–1425

    Article  CAS  Google Scholar 

  27. Zhou J, Chen Q, Lanske B, Fleming BC, Terek R, Wei X, Zhang G, Wang S, Li K, Wei L (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

    Article  Google Scholar 

  28. Lee BJ, Yang S, Kwon S, Choi KH, Kim W (2019) Association between metabolic syndrome and knee osteoarthritis: a cross-sectional nationwide survey study. J Rehabil Med 51:464–470

    PubMed  Google Scholar 

  29. Zhou X, Li J, Zhou Y, Yang Z, Yang H et al (2020) Down-regulated ciRS-7/up-regulated miR-7 axis aggravated cartilage degradation and autophagy defection by PI3K/AKT/mTOR activation mediated by IL-17A in osteoarthritis. Aging (Albany NY) 12:20163–20183

    Article  CAS  Google Scholar 

  30. Xu K, He Y, Moqbel SAA, Zhou X, Wu L, Bao J (2021) SIRT3 ameliorates osteoarthritis via regulating chondrocyte autophagy and apoptosis through the PI3K/Akt/mTOR pathway. Int J Biol Macromol 175:351–360

    Article  CAS  Google Scholar 

  31. Liu Y, Zhao D, Wang X, Dong Y, Ding F (2021) LncRNA KCNQ1OT1 attenuates osteoarthritic chondrocyte dysfunction via the miR-218-5p/PIK3C2A axis. Cell Tissue Res 385:115–126

    Article  CAS  Google Scholar 

  32. Han G, Zhang Y, Li H (2021) The Combination treatment of curcumin and probucol protects chondrocytes from tnf-alpha induced inflammation by enhancing autophagy and reducing apoptosis via the PI3K-Akt-mTOR pathway. Oxid Med Cell Longev 2021:5558066

    PubMed  PubMed Central  Google Scholar 

  33. Zhang Q, Lai S, Hou X, Cao W, Zhang Y, Zhang Z (2018) Protective effects of PI3K/Akt signal pathway induced cell autophagy in rat knee joint cartilage injury. Am J Transl Res 10:762–770

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Qureshi HY, Ahmad R, Sylvester J, Zafarullah M (2007) Requirement of phosphatidylinositol 3-kinase/Akt signaling pathway for regulation of tissue inhibitor of metalloproteinases-3 gene expression by TGF-beta in human chondrocytes. Cell Signal 19:1643–1651

    Article  CAS  Google Scholar 

  35. Zhang M, Zhou Q, Liang QQ, Li CG, Holz JD, Tang D, Sheu TJ, Li TF, Shi Q, Wang YJ (2009) IGF-1 regulation of type II collagen and MMP-13 expression in rat endplate chondrocytes via distinct signaling pathways. Osteoarthritis Cartilage 17:100–106

    Article  CAS  Google Scholar 

  36. Chen W, Zhao W, Yang A, Xu A, Wang H, Cong M, Liu T, Wang P, You H (2017) Integrated analysis of microRNA and gene expression profiles reveals a functional regulatory module associated with liver fibrosis. Gene 636:87–95

    Article  CAS  Google Scholar 

  37. Zhu M, Yang H, Chen Z, Xia X, Deng Q, Shen Y (2020) A cell-permeable peptide inhibitor of p55PIK signaling alleviates ocular inflammation in mouse models of uveitis. Exp Eye Res 199:108180

    Article  CAS  Google Scholar 

  38. Lv F, Yu Y, Wang G, Hu J, Zhang B, You W, Wang J (2015) Mechanisms by which the N-terminal 24 amino acids of the p55 regulatory subunit of phosphatidylinositol 3-kinase affect endotoxin-induced cytokine release in human keratinocytes. Mol Med Rep 11:3753–3759

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Orthopedics, The First People’s Hospital of Hangzhou, No.261, Huansha Road, Shangcheng District, Zhejiang Province, Hangzhou, 310000, China

    Lanjuan Shen, Cheng Ji & Hongping Yang

  2. Department of Orthopedics, Hangzhou Xiaoshan Hospital of Traditional Chinese Medicine, Zhejiang Province, Hangzhou City, China

    Jian Lin

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  1. Lanjuan Shen
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  2. Cheng Ji
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  3. Jian Lin
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  4. Hongping Yang
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Contributions

LS was responsible for drafting the manuscript. LS, CJ, and JL contributed to the analysis and interpretation of data. LS and HY contributed in the data collection. All authors read and approved the final manuscript.

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Correspondence to Hongping Yang.

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Written informed consent was obtained from patients with approval by the Institutional Review Board in The First People's Hospital of Hangzhou.

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Shen, L., Ji, C., Lin, J. et al. Regulation of circADAMTS6-miR-324-5p-PIK3R3 ceRNA pathway may be a novel mechanism of IL-1β-induced osteoarthritic chondrocytes. J Bone Miner Metab 40, 389–401 (2022). https://doi.org/10.1007/s00774-021-01308-0

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  • DOI: https://doi.org/10.1007/s00774-021-01308-0

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