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DJ-1-mediated p62 degradation delays intervertebral disc degeneration by inhibiting apoptosis of nucleus pulposus cells

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Abstract

Intervertebral disc degeneration (IDD) is the most important pathological basis of degenerative spinal diseases, for which effective interventions are still lacking. Oxidative stress is considered to be one of the leading pathological mechanisms contributing to IDD. However, the exact role of DJ-1 as an essential member of the antioxidant defense system in IDD is still unclear. Therefore, the aim of this study was to investigate the role played by DJ-1 in IDD and to reveal its potential molecular mechanisms. Western blot and immunohistochemical staining assays were performed to detect the expression of DJ-1 in degenerative nucleus pulposus cells (NPCs). After overexpression of DJ-1 in NPCs by lentiviral transfection, DCFH-DA and MitoSOX fluorescent probes were used to evaluate the levels of reactive oxygen species (ROS); while western blot, TUNEL staining, and Caspase-3 activity were used to assess apoptosis. Immunofluorescence staining was used to demonstrate the relationship between DJ-1 and p62. After inhibition of lysosomal degradation function with chloroquine, p62 degradation and apoptosis in DJ-1 overexpressing NPCs were further examined. In vivo, we assessed the therapeutic effect of upregulated DJ-1 on IDD by X-ray, MRI and Safranin O-Fast green staining. The protein expression of DJ-1 was significantly decreased in degenerated NPCs, accompanied by increased apoptosis. However, overexpression of DJ-1 significantly inhibited the elevated ROS levels and apoptosis in NPCs under oxidative stress. Mechanistically, our results showed that upregulation of DJ-1 promoted p62 degradation via the autophagic lysosomal pathway and that the protective effect of DJ-1 on NPCs under oxidative stress was partially mediated by promoting lysosomal pathway degradation of p62. Moreover, intradiscal injection of adeno-associated virus for overexpression of DJ-1 mitigated the progression of IDD in rats. This study reveals that DJ-1 maintains the homeostasis of NPCs by promoting the degradation of p62 through the autophagic lysosomal pathway, suggesting that DJ-1 is a promising new target for IDD intervention.

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Data availability

The datasets generated and/or analyzed during the current study are available in the manuscript and supplementary materials.

Abbreviations

IDD:

Intervertebral disc degeneration

NPCs:

Nucleus pulposus cells

ROS:

Reactive oxygen species

LBP:

Low back pain

NP:

Nucleus pulposus

AF:

Annulus fibrosus

DMEM/F12:

Dulbecco's Modified Eagle Medium: F-12

FBS:

Fetal bovine serum

TBHP:

Tert-butyl hydroperoxide

PBS:

Phosphate buffer saline

DAPI:

4′,6-Diamidino-2-phenylindole

RIPA:

Radio immunoprecipitation assay

PMSF:

Phenylmethanesulfonyl fluoride

BCA:

Bicinchoninic acid

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

PVDF:

Polyvinylidene difluoride

TBST:

Tris buffered saline tween 20

MRI:

Magnetic resonance imaging

DHI:

Disc height index

HE:

Hematoxylin–eosin

SO&FG:

Safranin O-Fast Green

DEPs:

Differentially expressed proteins

CQ:

Chloroquine

ECM:

Extracellular matrix

References

  1. (2020) Global burden of 369 diseases and injuries in 204 countries and territories, 1990–2019: a systematic analysis for the Global Burden of Disease Study 2019. Lancet (London, England) 396(10258):1204–1222

  2. Balagué F, Mannion AF, Pellisé F, Cedraschi C (2012) Non-specific low back pain. Lancet (London, England) 379(9814):482–491

    Article  PubMed  Google Scholar 

  3. Andersson GB (1999) Epidemiological features of chronic low-back pain. Lancet (London, England) 354(9178):581–585

    Article  CAS  PubMed  Google Scholar 

  4. Shirazi-Adl A, Taheri M, Urban JP (2010) Analysis of cell viability in intervertebral disc: effect of endplate permeability on cell population. J Biomech 43(7):1330–1336

    Article  CAS  PubMed  Google Scholar 

  5. Vergroesen PP, Kingma I, Emanuel KS, Hoogendoorn RJ, Welting TJ, van Royen BJ, van Dieën JH, Smit TH (2015) Mechanics and biology in intervertebral disc degeneration: a vicious circle. Osteoarthr Cartil 23(7):1057–1070

    Article  Google Scholar 

  6. Adams MA, Roughley PJ (2006) What is intervertebral disc degeneration, and what causes it? Spine 31(18):2151–2161

    Article  PubMed  Google Scholar 

  7. Mern DS, Beierfuß A, Thomé C, Hegewald AA (2014) Enhancing human nucleus pulposus cells for biological treatment approaches of degenerative intervertebral disc diseases: a systematic review. J Tissue Eng Regen Med 8(12):925–936

    Article  CAS  PubMed  Google Scholar 

  8. Newell N, Carpanen D, Evans JH, Pearcy MJ, Masouros SD (2019) Mechanical function of the nucleus pulposus of the intervertebral disc under high rates of loading. Spine 44(15):1035–1041

    Article  PubMed  Google Scholar 

  9. Vamvakas SS, Mavrogonatou E, Kletsas D (2017) Human nucleus pulposus intervertebral disc cells becoming senescent using different treatments exhibit a similar transcriptional profile of catabolic and inflammatory genes. Eur Spine J 26(8):2063–2071

    Article  PubMed  Google Scholar 

  10. Silagi ES, Shapiro IM, Risbud MV (2018) Glycosaminoglycan synthesis in the nucleus pulposus: dysregulation and the pathogenesis of disc degeneration. Matrix 71–72:368–379

    Article  Google Scholar 

  11. Lin J, Chen J, Zhang Z, Xu T, Shao Z, Wang X, Ding Y, Tian N, Jin H, Sheng S et al (2019) Luteoloside inhibits IL-1β-induced apoptosis and catabolism in nucleus pulposus cells and ameliorates intervertebral disk degeneration. Front Pharmacol 10:868

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Ding F, Shao ZW, Xiong LM (2013) Cell death in intervertebral disc degeneration. Apoptosis 18(7):777–785

    Article  PubMed  Google Scholar 

  13. Gruber HE, Hanley EN Jr (1998) Analysis of aging and degeneration of the human intervertebral disc. Comparison of surgical specimens with normal controls. Spine 23(7):751–757

    Article  CAS  PubMed  Google Scholar 

  14. Tang Z, Ju Y, Dai X, Ni N, Liu Y, Zhang D, Gao H, Sun H, Zhang J, Gu P (2021) HO-1-mediated ferroptosis as a target for protection against retinal pigment epithelium degeneration. Redox Biol 43:101971

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Kadenbach B, Ramzan R, Vogt S (2009) Degenerative diseases, oxidative stress and cytochrome c oxidase function. Trends Mol Med 15(4):139–147

    Article  CAS  PubMed  Google Scholar 

  16. Cao SS, Kaufman RJ (2014) Endoplasmic reticulum stress and oxidative stress in cell fate decision and human disease. Antioxid Redox Signal 21(3):396–413

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Navarro-Yepes J, Burns M, Anandhan A, Khalimonchuk O, del Razo LM, Quintanilla-Vega B, Pappa A, Panayiotidis MI, Franco R (2014) Oxidative stress, redox signaling, and autophagy: cell death versus survival. Antioxid Redox Signal 21(1):66–85

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Dalleau S, Baradat M, Guéraud F, Huc L (2013) Cell death and diseases related to oxidative stress: 4-hydroxynonenal (HNE) in the balance. Cell Death Differ 20(12):1615–1630

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Orrenius S, Gogvadze V, Zhivotovsky B (2007) Mitochondrial oxidative stress: implications for cell death. Annu Rev Pharmacol Toxicol 47:143–183

    Article  CAS  PubMed  Google Scholar 

  20. Song Y, Li S, Geng W, Luo R, Liu W, Tu J, Wang K, Kang L, Yin H, Wu X et al (2018) Sirtuin 3-dependent mitochondrial redox homeostasis protects against AGEs-induced intervertebral disc degeneration. Redox Biol 19:339–353

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Lin J, Zheng X, Zhang Z, Zhuge J, Shao Z, Huang C, Jin J, Chen X, Chen Y, Wu Y et al (2021) Inhibition of LRRK2 restores parkin-mediated mitophagy and attenuates intervertebral disc degeneration. Osteoarthr Cartil 29(4):579–591

    Article  Google Scholar 

  22. Xu WN, Zheng HL, Yang RZ, Liu T, Yu W, Zheng XF, Li B, Jiang SD, Jiang LS (2019) Mitochondrial NDUFA4L2 attenuates the apoptosis of nucleus pulposus cells induced by oxidative stress via the inhibition of mitophagy. Exp Mol Med 51(11):1–16

    Article  PubMed  Google Scholar 

  23. Irrcher I, Aleyasin H, Seifert EL, Hewitt SJ, Chhabra S, Phillips M, Lutz AK, Rousseaux MW, Bevilacqua L, Jahani-Asl A et al (2010) Loss of the Parkinson’s disease-linked gene DJ-1 perturbs mitochondrial dynamics. Hum Mol Genet 19(19):3734–3746

    Article  CAS  PubMed  Google Scholar 

  24. Kim RH, Smith PD, Aleyasin H, Hayley S, Mount MP, Pownall S, Wakeham A, You-Ten AJ, Kalia SK, Horne P et al (2005) Hypersensitivity of DJ-1-deficient mice to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyrindine (MPTP) and oxidative stress. Proc Natl Acad Sci USA 102(14):5215–5220

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Pfirrmann CW, Metzdorf A, Zanetti M, Hodler J, Boos N (2001) Magnetic resonance classification of lumbar intervertebral disc degeneration. Spine 26(17):1873–1878

    Article  CAS  PubMed  Google Scholar 

  26. Lin J, Zhuge J, Zheng X, Wu Y, Zhang Z, Xu T, Meftah Z, Xu H, Wu Y, Tian N et al (2020) Urolithin A-induced mitophagy suppresses apoptosis and attenuates intervertebral disc degeneration via the AMPK signaling pathway. Free Radic Biol Med 150:109–119

    Article  CAS  PubMed  Google Scholar 

  27. Han B, Zhu K, Li FC, Xiao YX, Feng J, Shi ZL, Lin M, Wang J, Chen QX (2008) A simple disc degeneration model induced by percutaneous needle puncture in the rat tail. Spine 33(18):1925–1934

    Article  PubMed  Google Scholar 

  28. Tam V, Chen P, Yee A, Solis N, Klein T, Kudelko M, Sharma R, Chan WC, Overall CM, Haglund L et al (2020) DIPPER, a spatiotemporal proteomics atlas of human intervertebral discs for exploring ageing and degeneration dynamics. eLife 9

  29. Cao G, Yang S, Cao J, Tan Z, Wu L, Dong F, Ding W, Zhang F (2022) The role of oxidative stress in intervertebral disc degeneration. Oxid Med Cell Longev 2022:2166817

    Article  PubMed  PubMed Central  Google Scholar 

  30. Billia F, Hauck L, Grothe D, Konecny F, Rao V, Kim RH, Mak TW (2013) Parkinson-susceptibility gene DJ-1/PARK7 protects the murine heart from oxidative damage in vivo. Proc Natl Acad Sci USA 110(15):6085–6090

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Gharbi N, Røise D, Førre JE, Edson AJ, Hushagen HA, Tronci V, Frøyset AK, Fladmark KE (2021) Reintroduction of DJ-1 in Müller Cells Inhibits Retinal Degeneration in the DJ-1 Deficient Retina. Antioxidants (Basel, Switzerland) 10(12)

  32. Chen Y, Lin J, Chen J, Huang C, Zhang Z, Wang J, Wang K, Wang X (2020) Mfn2 is involved in intervertebral disc degeneration through autophagy modulation. Osteoarthr Cartil 28(3):363–374

    Article  CAS  Google Scholar 

  33. Zheng G, Pan Z, Zhan Y, Tang Q, Zheng F, Zhou Y, Wu Y, Zhou Y, Chen D, Chen J et al (2019) TFEB protects nucleus pulposus cells against apoptosis and senescence via restoring autophagic flux. Osteoarthr Cartil 27(2):347–357

    Article  Google Scholar 

  34. Chan JY, Chan SH (2015) Activation of endogenous antioxidants as a common therapeutic strategy against cancer, neurodegeneration and cardiovascular diseases: a lesson learnt from DJ-1. Pharmacol Ther 156:69–74

    Article  CAS  PubMed  Google Scholar 

  35. Won KJ, Jung SH, Jung SH, Lee KP, Lee HM, Lee DY, Park ES, Kim J, Kim B (2014) DJ-1/park7 modulates vasorelaxation and blood pressure via epigenetic modification of endothelial nitric oxide synthase. Cardiovasc Res 101(3):473–481

    Article  CAS  PubMed  Google Scholar 

  36. Huang M, Chen S (2021) DJ-1 in neurodegenerative diseases: pathogenesis and clinical application. Prog Neurobiol 204:102114

    Article  CAS  PubMed  Google Scholar 

  37. Lin Z, Chen C, Yang D, Ding J, Wang G, Ren H (2021) DJ-1 inhibits microglial activation and protects dopaminergic neurons in vitro and in vivo through interacting with microglial p65. Cell Death Dis 12(8):715

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Baulac S, Lu H, Strahle J, Yang T, Goldberg MS, Shen J, Schlossmacher MG, Lemere CA, Lu Q, Xia W (2009) Increased DJ-1 expression under oxidative stress and in Alzheimer’s disease brains. Mol Neurodegener 4:12

    Article  PubMed  PubMed Central  Google Scholar 

  39. Pisani A, Martella G, Tscherter A, Costa C, Mercuri NB, Bernardi G, Shen J, Calabresi P (2006) Enhanced sensitivity of DJ-1-deficient dopaminergic neurons to energy metabolism impairment: role of Na+/K+ ATPase. Neurobiol Dis 23(1):54–60

    Article  CAS  PubMed  Google Scholar 

  40. Amatullah H, Maron-Gutierrez T, Shan Y, Gupta S, Tsoporis JN, Varkouhi AK, Teixeira Monteiro AP, He X, Yin J, Marshall JC et al (2021) Protective function of DJ-1/PARK7 in lipopolysaccharide and ventilator-induced acute lung injury. Redox Biol 38:101796

    Article  CAS  PubMed  Google Scholar 

  41. Dongworth RK, Mukherjee UA, Hall AR, Astin R, Ong SB, Yao Z, Dyson A, Szabadkai G, Davidson SM, Yellon DM et al (2014) DJ-1 protects against cell death following acute cardiac ischemia-reperfusion injury. Cell Death Dis 5(2):e1082

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Raninga PV, Di Trapani G, Vuckovic S, Tonissen KF (2016) Targeted knockdown of DJ-1 induces multiple myeloma cell death via KLF6 upregulation. Apoptosis 21(12):1422–1437

    Article  CAS  PubMed  Google Scholar 

  43. Zhang F, Yan Y, Peng W, Wang L, Wang T, Xie Z, Luo H, Zhang J, Dong W (2021) PARK7 promotes repair in early steroid-induced osteonecrosis of the femoral head by enhancing resistance to stress-induced apoptosis in bone marrow mesenchymal stem cells via regulation of the Nrf2 signaling pathway. Cell Death Dis 12(10):940

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Perlman H, Zhang X, Chen MW, Walsh K, Buttyan R (1999) An elevated bax/bcl-2 ratio corresponds with the onset of prostate epithelial cell apoptosis. Cell Death Differ 6(1):48–54

    Article  CAS  PubMed  Google Scholar 

  45. Yang B, Johnson TS, Thomas GL, Watson PF, Wagner B, Furness PN, El Nahas AM (2002) A shift in the Bax/Bcl-2 balance may activate caspase-3 and modulate apoptosis in experimental glomerulonephritis. Kidney Int 62(4):1301–1313

    Article  CAS  PubMed  Google Scholar 

  46. Klionsky DJ, Petroni G, Amaravadi RK, Baehrecke EH, Ballabio A, Boya P, Bravo-San Pedro JM, Cadwell K, Cecconi F, Choi AMK et al (2021) Autophagy in major human diseases. EMBO J 40(19):e108863

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Lee DH, Kim D, Kim ST, Jeong S, Kim JL, Shim SM, Heo AJ, Song X, Guo ZS, Bartlett DL et al (2018) PARK7 modulates autophagic proteolysis through binding to the N-terminally arginylated form of the molecular chaperone HSPA5. Autophagy 14(11):1870–1885

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Thomas KJ, McCoy MK, Blackinton J, Beilina A, van der Brug M, Sandebring A, Miller D, Maric D, Cedazo-Minguez A, Cookson MR (2011) DJ-1 acts in parallel to the PINK1/parkin pathway to control mitochondrial function and autophagy. Hum Mol Genet 20(1):40–50

    Article  CAS  PubMed  Google Scholar 

  49. Chen WT, Yang HB, Ke TW, Liao WL, Hung SY (2021) Serum DJ-1 Is a biomarker of colorectal cancer and DJ-1 activates mitophagy to promote colorectal cancer progression. Cancers 13(16)

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Acknowledgements

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Funding

This work was supported by the National Natural Science Foundation of China (Grant No. 82172480).

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Authors and Affiliations

  1. Department of Orthopaedics, Peking University Third Hospital, Haidian District, 49 North Garden Road, Beijing, 100191, China

    Jialiang Lin, Xuanqi Zheng, Qian Xiang, Yongzhao Zhao, Shuai Jiang, Zhuoran Sun, Dongwei Fan, Chuiguo Sun & Weishi Li

  2. Beijing Key Laboratory of Spinal Disease Research, Beijing, China

    Jialiang Lin, Xuanqi Zheng, Qian Xiang, Yongzhao Zhao, Shuai Jiang, Zhuoran Sun, Dongwei Fan, Chuiguo Sun & Weishi Li

  3. Engineering Research Center of Bone and Joint Precision Medicine, Ministry of Education, Beijing, China

    Jialiang Lin, Xuanqi Zheng, Qian Xiang, Yongzhao Zhao, Shuai Jiang, Zhuoran Sun, Dongwei Fan, Chuiguo Sun & Weishi Li

  4. Peking University Health Science Center, Beijing, China

    Jialiang Lin, Xuanqi Zheng, Qian Xiang & Yongzhao Zhao

  5. Trauma Medical Center, Department of Orthopedics Surgery, West China Hospital, West China Medical School, Sichuan University, Chengdu, China

    Zhencheng Xiong

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Contributions

JL: Conceptualization, Data curation, Formal Analysis, Validation, Writing – original draft; XZ: Investigation, Methodology, Data curation, Resources, Formal Analysis; ZX: Resources, Software, Visualization, Methodology; QX: Methodology, Investigation, Validation; YZ: Software, Formal Analysis; SJ: Data curation, Resources; ZS: Resources; DF: Formal Analysis, Methodology; CS: Resources; WL: Conceptualization, Funding acquisition, Supervision, Project administration, Writing – review & editing.

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Correspondence to Weishi Li.

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Human samples were used in strict accordance with The Code of Ethics of the World Medical Association (Declaration of Helsinki). All animal use in this study followed the guidelines of the Peking University Animal Care and Use Committee. Ethics approval was obtained from the Biomedical Ethics Committee of Peking University (No.LA2022421).

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Lin, J., Zheng, X., Xiong, Z. et al. DJ-1-mediated p62 degradation delays intervertebral disc degeneration by inhibiting apoptosis of nucleus pulposus cells. Apoptosis 28, 1357–1371 (2023). https://doi.org/10.1007/s10495-023-01862-0

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