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COX17 restricts renal fibrosis development by maintaining mitochondrial copper homeostasis and restoring complex IV activity

Acta Pharmacologica Sinica volume 44, pages 2091–2102 (2023)Cite this article

Abstract

Renal fibrosis relies on multiple proteins and cofactors in its gradual development. Copper is a cofactor of many enzymes involved in renal microenvironment homeostasis. We previously reported that intracellular copper imbalance occurred during renal fibrosis development and was correlated with fibrosis intensity. In this study, we investigated the molecular mechanisms of how copper affected renal fibrosis development. Unilateral ureteral obstruction (UUO) mice were used for in vivo study; rat renal tubular epithelial cells (NRK-52E) treated with TGF-β1 were adapted as an in vitro fibrotic model. We revealed that the accumulation of copper in mitochondria, rather than cytosol, was responsible for mitochondrial dysfunction, cell apoptosis and renal fibrosis in both in vivo and in vitro fibrotic models. Furthermore, we showed that mitochondrial copper overload directly disrupted the activity of respiratory chain complex IV (cytochrome c oxidase), but not complex I, II and III, which hampered respiratory chain and disrupted mitochondrial functions, eventually leading to fibrosis development. Meanwhile, we showed that COX17, the copper chaperone protein, was significantly upregulated in the mitochondria of fibrotic kidneys and NRK-52E cells. Knockdown of COX17 aggravated mitochondrial copper accumulation, inhibited complex IV activity, augmented mitochondrial dysfunction and led to cell apoptosis and renal fibrosis, whereas overexpression of COX17 could discharge copper from mitochondria and protect mitochondrial function, alleviating renal fibrosis. In conclusion, copper accumulation in mitochondria blocks complex IV activity and induces mitochondrial dysfunction. COX17 plays a pivotal role in maintaining mitochondrial copper homeostasis, restoring complex IV activity, and ameliorating renal fibrosis.

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Fig. 1: COX17 is upregulated in renal interstitial fibrosis.
Fig. 2: Downregulation of COX17 in vivo aggravates renal fibrosis.
Fig. 3: Downregulation of COX17 aggravates TGF-β1 induced-mitochondrial dysfunction and cell apoptosis.
Fig. 4: Overexpression of COX17 mitigates mitochondrial dysfunction and cell apoptosis.
Fig. 5: COX17 reduces mitochondrial copper content and restores respiratory chain complex IV activity.
Fig. 6: Alleviating mitochondrial copper overload improves mitochondrial dysfunction, cell apoptosis and renal fibrosis.
Fig. 7: Diagram depicts the role of COX17 in renal fibrosis.

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References

  1. Panizo S, Martinez-Arias L, Alonso-Montes C, Cannata P, Martin-Carro B, Fernandez-Martin JL, et al. Fibrosis in chronic kidney disease: pathogenesis and consequences. Int J Mol Sci. 2021;22:408–26.

  2. Lutsenko S. Human copper homeostasis: a network of interconnected pathways. Curr Opin Chem Biol. 2010;14:211–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Kim BE, Nevitt T, Thiele DJ. Mechanisms for copper acquisition, distribution and regulation. Nat Chem Biol. 2008;4:176–85.

    Article  CAS  PubMed  Google Scholar 

  4. Janssen R, de Brouwer B, von der Thusen JH, Wouters EFM. Copper as the most likely pathogenic divergence factor between lung fibrosis and emphysema. Med Hypotheses. 2018;120:49–54.

    Article  CAS  PubMed  Google Scholar 

  5. Yadav A, Kumar L, Misra N, Deepak U, Shiv GC. Kumar, Estimation of serum zinc, copper, and iron in the patients of oral submucous fibrosis. Natl J Maxillofac Surg. 2015;6:190–3.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Dirksen K, Fieten H. Canine copper-associated hepatitis. Vet Clin North Am Small Anim Pract. 2017;47:631–44.

    Article  PubMed  Google Scholar 

  7. Tadakamadla J, Kumar S, Gp M. Evaluation of serum copper and iron levels among oral submucous fibrosis patients. Med Oral Patol Oral Cir Bucal. 2011;16:e870–3.

    Article  PubMed  Google Scholar 

  8. Brewer GJ, Dick R, Ullenbruch MR, Jin H, Phan SH. Inhibition of key cytokines by tetrathiomolybdate in the bleomycin model of pulmonary fibrosis. J Inorg Biochem. 2004;98:2160–7.

    Article  CAS  PubMed  Google Scholar 

  9. Niu YY, Zhang YY, Zhu Z, Zhang XQ, Liu X, Zhu SY, et al. Elevated intracellular copper contributes a unique role to kidney fibrosis by lysyl oxidase mediated matrix crosslinking. Cell Death Dis. 2020;11:211.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Zhang F, Pracheil T, Thornton J, Liu Z. Adenosine Triphosphate (ATP) is a candidate signaling molecule in the mitochondria-to-nucleus retrograde response pathway. Genes (Basel). 2013;4:86–100.

    Article  PubMed  Google Scholar 

  11. Zischka H, Einer C. Mitochondrial copper homeostasis and its derailment in Wilson disease. Int J Biochem Cell Biol. 2018;102:71–5.

    Article  CAS  PubMed  Google Scholar 

  12. Kang HM, Ahn SH, Choi P, Ko YA, Han SH, Chinga F, et al. Defective fatty acid oxidation in renal tubular epithelial cells has a key role in kidney fibrosis development. Nat Med. 2015;21:37–46.

    Article  CAS  PubMed  Google Scholar 

  13. Sivitz WI, Yorek MA. Mitochondrial dysfunction in diabetes: from molecular mechanisms to functional significance and therapeutic opportunities. Antioxid Redox Signal. 2010;12:537–77.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Sharma K, Karl B, Mathew AV, Gangoiti JA, Wassel CL, Saito R, et al. Metabolomics reveals signature of mitochondrial dysfunction in diabetic kidney disease. J Am Soc Nephrol. 2013;24:1901–12.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Johnson A, DiPietro LA. Apoptosis and angiogenesis: an evolving mechanism for fibrosis. FASEB J. 2013;27:3893–901.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Livingston MJ, Ding HF, Huang S, Hill JA, Yin XM, Dong Z. Persistent activation of autophagy in kidney tubular cells promotes renal interstitial fibrosis during unilateral ureteral obstruction. Autophagy. 2016;12:976–98.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Hsu PY, Yen HH, Yang TH, Su CC. Tetrathiomolybdate, a copper chelator inhibited imiquimod-induced skin inflammation in mice. J Dermatol Sci. 2018;92:30–7.

    Article  CAS  PubMed  Google Scholar 

  18. Brewer GJ, Ullenbruch MR, Dick R, Olivarez L, Phan SH. Tetrathiomolybdate therapy protects against bleomycin-induced pulmonary fibrosis in mice. J Lab Clin Med. 2003;141:210–6.

    Article  CAS  PubMed  Google Scholar 

  19. Zhu H, Jiang W, Zhao H, He C, Tang X, Xu S, et al. PSTPIP2 inhibits cisplatin-induced acute kidney injury by suppressing apoptosis of renal tubular epithelial cells. Cell Death Dis. 2020;11:1057.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Yu Y, Wang Y, Niu Y, Fu L, Chin YE, Yu C. Leukemia inhibitory factor attenuates renal fibrosis through Stat3-miR-29c. Am J Physiol Ren Physiol. 2015;309:F595–603.

    Article  CAS  Google Scholar 

  21. Han SH, Wu MY, Nam BY, Park JT, Yoo TH, Kang SW, et al. PGC-1alpha protects from notch-induced kidney fibrosis development. J Am Soc Nephrol. 2017;28:3312–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics. 2014;30:2114–20.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Dobin A, Davis CA, Schlesinger F, Drenkow J, Zaleski C, Jha S, et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics. 2013;29:15–21.

    Article  CAS  PubMed  Google Scholar 

  24. Feng J, Meyer CA, Wang Q, Liu JS, Shirley Liu X, Zhang Y. GFOLD: a generalized fold change for ranking differentially expressed genes from RNA-seq data. Bioinformatics. 2012;28:2782–8.

    Article  CAS  PubMed  Google Scholar 

  25. Feng L, Wang J, Li H, Luo X, Li J. A novel absolute quantitative imaging strategy of iron, copper and zinc in brain tissues by Isotope Dilution Laser Ablation ICP-MS. Anal Chim Acta. 2017;984:66–75.

    Article  CAS  PubMed  Google Scholar 

  26. Li SW, Shao YZ, Zhao HJ, Wang Y, Li JL, Xing MW. Analysis of 28 trace elements in the blood and serum antioxidant status in chickens under arsenic and/or copper exposure. Environ Sci Pollut Res Int. 2017;24:27303–13.

    Article  CAS  PubMed  Google Scholar 

  27. Wang Y, Lu M, Xiong L, Fan J, Zhou Y, Li H, et al. Drp1-mediated mitochondrial fission promotes renal fibroblast activation and fibrogenesis. Cell Death Dis. 2020;11:29.

    Article  PubMed  PubMed Central  Google Scholar 

  28. Li L, Kang H, Zhang Q, D’Agati VD, Al-Awqati Q, Lin F. FoxO3 activation in hypoxic tubules prevents chronic kidney disease. J Clin Invest. 2019;129:2374–89.

    Article  PubMed  PubMed Central  Google Scholar 

  29. Jumper J, Evans R, Pritzel A, Green T, Figurnov M, Ronneberger O, et al. Highly accurate protein structure prediction with AlphaFold. Nature. 2021;596:583–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Kadenbach B, Huttemann M. The subunit composition and function of mammalian cytochrome c oxidase. Mitochondrion. 2015;24:64–76.

    Article  CAS  PubMed  Google Scholar 

  31. Van Der Spoel D, Lindahl E, Hess B, Groenhof G, Mark AE, Berendsen HJ. GROMACS: fast, flexible, and free. J Comput Chem. 2005;26:1701–18.

    Article  PubMed  Google Scholar 

  32. Schmid N, Eichenberger AP, Choutko A, Riniker S, Winger M, Mark AE, et al. Definition and testing of the GROMOS force-field versions 54A7 and 54B7. Eur Biophys J. 2011;40:843–56.

    Article  CAS  PubMed  Google Scholar 

  33. Zhang L, Wang K, Lei Y, Li Q, Nice EC, Huang C. Redox signaling: potential arbitrator of autophagy and apoptosis in therapeutic response. Free Radic Biol Med. 2015;89:452–65.

    Article  CAS  PubMed  Google Scholar 

  34. Wikstrom M, Krab K, Sharma V. Oxygen activation and energy conservation by cytochrome c oxidase. Chem Rev. 2018;118:2469–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Timon-Gomez A, Nyvltova E, Abriata LA, Vila AJ, Hosler J, Barrientos A. Mitochondrial cytochrome c oxidase biogenesis: recent developments. Semin Cell Dev Biol. 2018;76:163–78.

    Article  CAS  PubMed  Google Scholar 

  36. Nevitt T, Ohrvik H, Thiele DJ. Charting the travels of copper in eukaryotes from yeast to mammals. Biochim Biophys Acta. 2012;1823:1580–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Maxfield AB, Heaton DN, Winge DR. Cox17 is functional when tethered to the mitochondrial inner membrane. J Biol Chem. 2004;279:5072–80.

    Article  CAS  PubMed  Google Scholar 

  38. Wang B, Dong D, Kang YJ. Copper chaperone for superoxide dismutase-1 transfers copper to mitochondria but does not affect cytochrome c oxidase activity. Exp Biol Med (Maywood). 2013;238:1017–23.

    Article  PubMed  Google Scholar 

  39. Singh RP, Jeyaraju DV, Voisin V, Hurren R, Xu C, Hawley JR, et al. Disrupting mitochondrial copper distribution inhibits leukemic stem cell self-renewal. Cell Stem Cell. 2020;26:926–37.

    Article  CAS  PubMed  Google Scholar 

  40. Ge EJ, Bush AI, Casini A, Cobine PA, Cross JR, DeNicola GM, et al. Connecting copper and cancer: from transition metal signalling to metalloplasia. Nat Rev Cancer. 2022;22:102–13.

    Article  CAS  PubMed  Google Scholar 

  41. Leary SC. Redox regulation of SCO protein function: controlling copper at a mitochondrial crossroad. Antioxid Redox Signal. 2010;13:1403–16.

    Article  CAS  PubMed  Google Scholar 

  42. Sokol RJ, Devereaux MW, O’Brien K, Khandwala RA, Loehr JP. Abnormal hepatic mitochondrial respiration and cytochrome C oxidase activity in rats with long-term copper overload. Gastroenterology. 1993;105:178–87.

    Article  CAS  PubMed  Google Scholar 

  43. Roberts EA, Robinson BH, Yang S. Mitochondrial structure and function in the untreated Jackson toxic milk (tx-j) mouse, a model for Wilson disease. Mol Genet Metab. 2008;93:54–65.

    Article  CAS  PubMed  Google Scholar 

  44. Zischka H, Lichtmannegger J. Pathological mitochondrial copper overload in livers of Wilson’s disease patients and related animal models. Ann N Y Acad Sci. 2014;1315:6–15.

    Article  CAS  PubMed  Google Scholar 

  45. Tsvetkov P, Coy S, Petrova B, Dreishpoon M, Verma A, Abdusamad M, et al. Copper induces cell death by targeting lipoylated TCA cycle proteins. Science. 2022;375:1254–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Funding

This study was supported by the National Natural Science Foundation of China (No 81873609, 82170696, 82000673).

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Authors and Affiliations

  1. Department of Nephrology, Tongji Hospital, School of Medicine, Tongji University, Shanghai, 200092, China

    Sai-ya Zhu, Wen-qian Zhou, Yang-yang Niu, Chao Zheng, Xi Liu, Ying-ying Zhang & Chen Yu

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Contributions

CY, SYZ and YYN designed the study; SYZ performed the experiments; SYZ, WQZ and XL analysed and interpreted the results; SYZ wrote and edited the manuscript. WQZ and CZ assisted with the main experiments. YYZ provided essential reagents and techniques for this study and reviewed the manuscript. CY, SYZ and YYN approved the final version of the manuscript. CY conceived and supervised the study.

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Correspondence to Chen Yu.

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Zhu, Sy., Zhou, Wq., Niu, Yy. et al. COX17 restricts renal fibrosis development by maintaining mitochondrial copper homeostasis and restoring complex IV activity. Acta Pharmacol Sin 44, 2091–2102 (2023). https://doi.org/10.1038/s41401-023-01098-3

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