Advertisement

Extracellular vesicles derived from mesenchymal stem cells confer protection against intervertebral disc degeneration through a microRNA-217-dependent mechanism

  • Author Footnotes
    a Yingjie Hao and Guangduo Zhu contributed equally to this work.
    Y. Hao
    Correspondence
    Address correspondence and reprint requests to: Y. Hao, Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, No. 1, Jianshe East Road, Zhengzhou 450052, Henan Province, PR China. Tel./Fax: 86-0371-67967206.
    Footnotes
    a Yingjie Hao and Guangduo Zhu contributed equally to this work.
    Affiliations
    Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, PR China
    Search for articles by this author
  • Author Footnotes
    a Yingjie Hao and Guangduo Zhu contributed equally to this work.
    G. Zhu
    Footnotes
    a Yingjie Hao and Guangduo Zhu contributed equally to this work.
    Affiliations
    Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, PR China
    Search for articles by this author
  • L. Yu
    Affiliations
    Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, PR China
    Search for articles by this author
  • Z. Ren
    Affiliations
    Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, PR China
    Search for articles by this author
  • P. Zhang
    Affiliations
    Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, PR China
    Search for articles by this author
  • J. Zhu
    Affiliations
    Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, PR China
    Search for articles by this author
  • S. Cao
    Affiliations
    Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, PR China
    Search for articles by this author
  • Author Footnotes
    a Yingjie Hao and Guangduo Zhu contributed equally to this work.
Published:August 27, 2022DOI:https://doi.org/10.1016/j.joca.2022.08.009

      Summary

      Objective

      Extracellular vesicles released by mesenchymal stem cells (MSC-EVs) can be applied to alleviate intervertebral disc degeneration (IVDD) by curbing apoptosis of nucleus pulposus cells (NPCs). The current study aims to evaluate the effect of MSC-EVs on NPC apoptosis and IVDD and the related regulatory mechanisms involving microRNA (miR)-217.

      Method

      Expression of miR-217 was examined in tumor necrosis factor-α (TNF-α)-induced NPCs and MSC-EVs, followed by identification in the relationship between miR-217, enhancer of zeste homolog 2 (EZH2) and forkhead box O-3 (FOXO3). After isolation of EVs from MSCs and subsequent co-culture with NPCs, we assessed effects of miR-217 on NPC viability, autophagy, senescence and apoptosis along with extracellular matrix (ECM) degradation. Further in vivo experiments were conducted in rat models of IVDD to substantiate the effect of miR-217 on IVDD.

      Results

      Poor miR-217 expression was found in TNF-α-induced NPCs, while high miR-217 expression was identified in MSC-EVs (P < 0.05). MSC-EVs transferred miR-217 to NPCs and increased its expression, thus attenuating NPC apoptosis and ECM degradation (elevated collagen II and aggrecan but reduced MMP13 and ADAMTS5) (P < 0.05). miR-217 targeted EZH2, and EZH2 bound to the FOXO3 promoter and consequently downregulated its expression. FOXO3 restrained NPC apoptosis and ECM degradation by stimulating cell autophagy (P < 0.05). Furthermore, in vivo experimental results confirmed the suppressive role of miR-217 shuttled by MSC-EVs in IVDD.

      Conclusion

      Overall, the delivery of miR-217 may be a novel mechanism underlying the effect of MSC-EVs on NPC apoptosis and ECM degradation following IVDD.

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Osteoarthritis and Cartilage
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Francisco V.
        • Pino J.
        • Gonzalez-Gay M.A.
        • Lago F.
        • Karppinen J.
        • Tervonen O.
        • et al.
        A new immunometabolic perspective of intervertebral disc degeneration.
        Nat Rev Rheumatol. 2022; 18: 47-60
        • Willems N.
        • Tellegen A.R.
        • Bergknut N.
        • Creemers L.B.
        • Wolfswinkel J.
        • Freudigmann C.
        • et al.
        Inflammatory profiles in canine intervertebral disc degeneration.
        BMC Vet Res. 2016; 12: 10
        • Gao B.
        • Jiang B.
        • Xing W.
        • Xie Z.
        • Luo Z.
        • Zou W.
        Discovery and application of postnatal nucleus pulposus progenitors essential for intervertebral disc homeostasis and degeneration.
        Adv Sci (Weinh). 2022; e2104888
        • Zhou Z.M.
        • Bao J.P.
        • Peng X.
        • Gao J.W.
        • Vlf C.
        • Zhang C.
        • et al.
        Small extracellular vesicles from hypoxic mesenchymal stem cells alleviate intervertebral disc degeneration by delivering miR-17-5p.
        Acta Biomater. 2022; 140: 641-658
        • Wen T.
        • Wang H.
        • Li Y.
        • Lin Y.
        • Zhao S.
        • Liu J.
        • et al.
        Bone mesenchymal stem cell-derived extracellular vesicles promote the repair of intervertebral disc degeneration by transferring microRNA-199a.
        Cell Cycle. 2021; 20: 256-270
        • Liao Z.
        • Li S.
        • Lu S.
        • Liu H.
        • Li G.
        • Ma L.
        • et al.
        Metformin facilitates mesenchymal stem cell-derived extracellular nanovesicles release and optimizes therapeutic efficacy in intervertebral disc degeneration.
        Biomaterials. 2021; 274120850
        • Wang C.
        • Cui L.
        • Gu Q.
        • Guo S.
        • Zhu B.
        • Liu X.
        • et al.
        The mechanism and function of miRNA in intervertebral disc degeneration.
        Orthop Surg. 2022; 14: 463-471
        • Li Z.
        • Yu X.
        • Shen J.
        • Chan M.T.
        • Wu W.K.
        MicroRNA in intervertebral disc degeneration.
        Cell Prolif. 2015; 48: 278-283
        • Liao Z.
        • Liu H.
        • Ma L.
        • Lei J.
        • Tong B.
        • Li G.
        • et al.
        Engineering extracellular vesicles restore the impaired cellular uptake and attenuate intervertebral disc degeneration.
        ACS Nano. 2021; 15: 14709-14724
        • Rani S.
        • Ryan A.E.
        • Griffin M.D.
        • Ritter T.
        Mesenchymal stem cell-derived extracellular vesicles: toward cell-free therapeutic applications.
        Mol Ther. 2015; 23: 812-823
        • Xue J.
        • Liu Y.
        • Luo F.
        • Lu X.
        • Xu H.
        • Liu X.
        • et al.
        Circ100284, via miR-217 regulation of EZH2, is involved in the arsenite-accelerated cell cycle of human keratinocytes in carcinogenesis.
        Biochim Biophys Acta, Mol Basis Dis. 2017; 1863: 753-763
        • Jiang C.
        • Guo Q.
        • Jin Y.
        • Xu J.J.
        • Sun Z.M.
        • Zhu D.C.
        • et al.
        Inhibition of EZH2 ameliorates cartilage endplate degeneration and attenuates the progression of intervertebral disc degeneration via demethylation of Sox-9.
        EBioMedicine. 2019; 48: 619-629
        • Gong C.
        • Yao S.
        • Gomes A.R.
        • Man E.P.
        • Lee H.J.
        • Gong G.
        • et al.
        BRCA1 positively regulates FOXO3 expression by restricting FOXO3 gene methylation and epigenetic silencing through targeting EZH2 in breast cancer.
        Oncogenesis. 2016; 5: e214
        • Morris B.J.
        • Willcox D.C.
        • Donlon T.A.
        • Willcox B.J.
        FOXO3: a major gene for human longevity--A mini-review.
        Gerontology. 2015; 61: 515-525
        • Gangadaran P.
        • Rajendran R.L.
        • Lee H.W.
        • Kalimuthu S.
        • Hong C.M.
        • Jeong S.Y.
        • et al.
        Extracellular vesicles from mesenchymal stem cells activates VEGF receptors and accelerates recovery of hindlimb ischemia.
        J Contr Release. 2017; 264: 112-126
        • Wei F.
        • Ma C.
        • Zhou T.
        • Dong X.
        • Luo Q.
        • Geng L.
        • et al.
        Exosomes derived from gemcitabine-resistant cells transfer malignant phenotypic traits via delivery of miRNA-222-3p.
        Mol Cancer. 2017; 16: 132
        • Livak K.J.
        • Schmittgen T.D.
        Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.
        Methods. 2001; 25: 402-408
        • Ge J.
        • Zhou Q.
        • Niu J.
        • Wang Y.
        • Yan Q.
        • Wu C.
        • et al.
        Melatonin protects intervertebral disc from degeneration by improving cell survival and function via activation of the ERK1/2 signaling pathway.
        Oxid Med Cell Longev. 2019; 20195120275
        • Chauvin M.
        • Lebrault C.
        • Duvaldestin P.
        The neuromuscular blocking effect of vecuronium on the human diaphragm.
        Anesth Analg. 1987; 66: 117-122
        • Xiang Q.
        • Kang L.
        • Wang J.
        • Liao Z.
        • Song Y.
        • Zhao K.
        • et al.
        CircRNA-CIDN mitigated compression loading-induced damage in human nucleus pulposus cells via miR-34a-5p/SIRT1 axis.
        EBioMedicine. 2020; 53102679
        • Sun Y.
        • Shi X.
        • Peng X.
        • Li Y.
        • Ma H.
        • Li D.
        • et al.
        MicroRNA-181a exerts anti-inflammatory effects via inhibition of the ERK pathway in mice with intervertebral disc degeneration.
        J Cell Physiol. 2020; 235: 2676-2686
        • Lai A.
        • Gansau J.
        • Gullbrand S.E.
        • Crowley J.
        • Cunha C.
        • Dudli S.
        • et al.
        Development of a standardized histopathology scoring system for intervertebral disc degeneration in rat models: an initiative of the ORS spine section.
        JOR Spine. 2021; 4: e1150
        • Cheng X.
        • Zhang L.
        • Zhang K.
        • Zhang G.
        • Hu Y.
        • Sun X.
        • et al.
        Circular RNA VMA21 protects against intervertebral disc degeneration through targeting miR-200c and X linked inhibitor-of-apoptosis protein.
        Ann Rheum Dis. 2018; 77: 770-779
        • Wu J.
        • He J.
        • Tian X.
        • Li H.
        • Wen Y.
        • Shao Q.
        • et al.
        Upregulation of miRNA-9-5p promotes angiogenesis after traumatic brain injury by inhibiting ptch-1.
        Neuroscience. 2020; 440: 160-174
        • Jiang Q.
        • Geng X.
        • Warren J.
        • Eugene Paul Cosky E.
        • Kaura S.
        • Stone C.
        • et al.
        Hypoxia inducible factor-1alpha (HIF-1alpha) mediates NLRP3 inflammasome-dependent-pyroptotic and apoptotic cell death following ischemic stroke.
        Neuroscience. 2020; 448: 126-139
        • Cui S.
        • Zhang L.
        microRNA-129-5p shuttled by mesenchymal stem cell-derived extracellular vesicles alleviates intervertebral disc degeneration via blockade of LRG1-mediated p38 MAPK activation.
        J Tissue Eng. 2021; 1220417314211021679
        • DiStefano T.J.
        • Vaso K.
        • Danias G.
        • Chionuma H.N.
        • Weiser J.R.
        • Iatridis J.C.
        Extracellular vesicles as an emerging treatment option for intervertebral disc degeneration: therapeutic potential, translational pathways, and regulatory considerations.
        Adv Healthc Mater. 2022; 11e2100596
        • Mensa E.
        • Guescini M.
        • Giuliani A.
        • Bacalini M.G.
        • Ramini D.
        • Corleone G.
        • et al.
        Small extracellular vesicles deliver miR-21 and miR-217 as pro-senescence effectors to endothelial cells.
        J Extracell Vesicles. 2020; 91725285
        • Lu X.D.
        • Liu Y.R.
        • Zhang Z.Y.
        Matrilin-3 alleviates extracellular matrix degradation of nucleus pulposus cells via induction of IL-1 receptor antagonist.
        Eur Rev Med Pharmacol Sci. 2020; 24: 5231-5241
        • Cheng X.
        • Zhang G.
        • Zhang L.
        • Hu Y.
        • Zhang K.
        • Sun X.
        • et al.
        Mesenchymal stem cells deliver exogenous miR-21 via exosomes to inhibit nucleus pulposus cell apoptosis and reduce intervertebral disc degeneration.
        J Cell Mol Med. 2018; 22: 261-276
        • Chen D.L.
        • Zhang D.S.
        • Lu Y.X.
        • Chen L.Z.
        • Zeng Z.L.
        • He M.M.
        • et al.
        microRNA-217 inhibits tumor progression and metastasis by downregulating EZH2 and predicts favorable prognosis in gastric cancer.
        Oncotarget. 2015; 6: 10868-10879
        • Ma K.G.
        • Shao Z.W.
        • Yang S.H.
        • Wang J.
        • Wang B.C.
        • Xiong L.M.
        • et al.
        Autophagy is activated in compression-induced cell degeneration and is mediated by reactive oxygen species in nucleus pulposus cells exposed to compression.
        Osteoarthritis Cartilage. 2013; 21: 2030-2038
        • Alvarez-Garcia O.
        • Matsuzaki T.
        • Olmer M.
        • Miyata K.
        • Mokuda S.
        • Sakai D.
        • et al.
        FOXO are required for intervertebral disk homeostasis during aging and their deficiency promotes disk degeneration.
        Aging Cell. 2018; 17e12800
        • Wang Y.
        • Yang Y.
        • Zuo R.
        • Wu J.
        • Zhang C.
        • Li C.
        • et al.
        FOXO3 protects nucleus pulposus cells against apoptosis under nutrient deficiency via autophagy.
        Biochem Biophys Res Commun. 2020; 524: 756-763