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Deubiquitinase OTUD6A in macrophages promotes intestinal inflammation and colitis via deubiquitination of NLRP3

Cell Death & Differentiation (2023)Cite this article

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Abstract

Inflammatory bowel disease (IBD) is a chronic inflammatory disorder of the gastrointestinal tract, which has been shown to increase the incidence of colorectal cancer. Recent studies have highlighted the role of ubiquitination, a post-translational modification, in the occurrence and development of colonic inflammation. Ovarian tumor deubiquitinase 6 A (OTUD6A) is a deubiquitinating enzyme, which regulates cell proliferation and tumorigenesis. In this study, we investigated the expression and role of OTUD6A in IBD. Wide-type or Otud6a−/− mice were used to develop dextran sodium sulfate (DSS)- or 2,6,4-trinitrobenzene sulfonic acid (TNBS)-induced colitis model, as well as azoxymethane (AOM)/DSS-induced colitis-associated cancer model. Bone marrow-derived macrophages (BMDMs) were isolated from wild-type and Otud6a−/− mice to dissect molecular mechanisms. Our data show that OTUD6A deficiency attenuated DSS or TNBS-induced colitis, as well as AOM/DSS-induced colitis-related colon cancer in vivo. Bone marrow transplantation experiments further revealed that OTUD6A in myeloid cells was responsible for exacerbation of DSS-induced colitis. Mechanistically, OTUD6A directly bound to NACHT domain of NLRP3 inflammasome and selectively cleaved K48-linked polyubiquitin chains from NLRP3 at K430 and K689 to enhance the stability of NLRP3, leading to increased IL-1β level and inflammation. Taken together, our research identifies a new function of OTUD6A in the pathogenesis of colitis by promoting NLRP3 inflammasome activation, suggesting that OTUD6A could be a potential target for the treatment of IBD.

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Fig. 1: OTUD6A deficiency attenuated colonic injury induced by colitis.
Fig. 2: OTUD6A deficiency decreases the inflammatory infiltration and production of proinflammatory cytokines.
Fig. 3: Deficiency of OTUD6A in myeloid cells alleviates DSS-induced colitis.
Fig. 4: OTUD6A deficiency suppresses AOM/DSS-induced colon cancer.
Fig. 5: Inhibition of OTUD6A blocks the activation of NLRP3 inflammasome.
Fig. 6: OTUD6A directly interacts with NLRP3.
Fig. 7: OTUD6A deubiquitinates NLRP3.
Fig. 8: OTUD6A stabilizes NLRP3 by cleaving its polyubiquitin chain at K430 and K689.

Data availability

All data needed to evaluate the conclusions in this study are presented in this manuscript or the supplementary information. The materials described in this study are either commercially available or available upon reasonable request from the corresponding authors.

References

  1. Na YR, Stakenborg M, Seok SH, Matteoli G. Macrophages in intestinal inflammation and resolution: a potential therapeutic target in IBD. Nat Rev Gastroenterol Hepatol. 2019;16:531–43.

    Article  CAS  PubMed  Google Scholar 

  2. Wallace KL, Zheng LB, Kanazawa Y, Shih DQ. Immunopathology of inflammatory bowel disease. World J Gastroenterol. 2014;20:6–21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Roda G, Ng SC, Kotze PG, Argollo M, Panaccione R, Spinelli A, et al. Crohn’s disease. Nat Rev Dis Primers 2020;6:22.

    Article  PubMed  Google Scholar 

  4. Kobayashi T, Siegmund B, Le Berre C, Wei SC, Ferrante M, Shen B, et al. Ulcerative colitis. Nat Rev Dis Primers 2020;6:74.

    Article  PubMed  Google Scholar 

  5. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2021;71:209–49.

    Article  PubMed  Google Scholar 

  6. Siegel RL, Miller KD, Sauer AG, Fedewa SA, Butterly LF, Anderson JC, et al. Colorectal cancer statistics, 2020. CA Cancer J Clin. 2020;70:145–64.

    Article  PubMed  Google Scholar 

  7. Faye AS, Holmer AK, Axelrad JE. Cancer in inflammatory bowel disease. Gastroenterol Clin North Am. 2022;51:649–66.

    Article  PubMed  Google Scholar 

  8. Song Y, Yuan M, Xu Y, Xu H. Tackling inflammatory bowel diseases: targeting proinflammatory cytokines and lymphocyte homing. Pharmaceuticals 2022;15:1080.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Ruan J, Schluter D, Naumann M, Waisman A, Wang X. Ubiquitin-modifying enzymes as regulators of colitis. Trends Mol Med. 2022;28:304–18.

    Article  CAS  PubMed  Google Scholar 

  10. Popovic D, Vucic D, Dikic I. Ubiquitination in disease pathogenesis and treatment. Nat Med. 2014;20:1242–53.

    Article  CAS  PubMed  Google Scholar 

  11. Clague MJ, Barsukov I, Coulson JM, Liu H, Rigden DJ, Urbe S. Deubiquitylases from genes to organism. Physiol Rev. 2013;93:1289–315.

    Article  CAS  PubMed  Google Scholar 

  12. Bednash JS, Mallampalli RK. Regulation of inflammasomes by ubiquitination. Cell Mol Immunol. 2016;13:722–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Lei H, Yang L, Xu HZ, Wang ZT, Li XY, Liu M, et al. Ubiquitin-specific protease 47 regulates intestinal inflammation through deubiquitination of TRAF6 in epithelial cells. Sci China-Life Sci. 2022;65:1624–35.

    Article  CAS  PubMed  Google Scholar 

  14. Wu B, Qiang LH, Zhang Y, Fu YS, Zhao MY, Lei ZH, et al. The deubiquitinase OTUD1 inhibits colonic inflammation by suppressing RIPK1-mediated NE-kappa B signaling. Cell Mol Immunol. 2022;19:276–89.

    Article  CAS  PubMed  Google Scholar 

  15. Shi L, Liu J, Peng YH, Zhang JF, Dai XP, Zhang SX, et al. Deubiquitinase OTUD6A promotes proliferation of cancer cells via regulating Drp1 stability and mitochondrial fission. Mol Oncol. 2020;14:3169–83.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Peng YH, Liu J, Wang Z, Cui CP, Zhang TT, Zhang SX, et al. Prostate-specific oncogene OTUD6A promotes prostatic tumorigenesis via deubiquitinating and stabilizing c-Myc. Cell Death Differ. 2022;29:1730–43.

    Article  CAS  PubMed  Google Scholar 

  17. Zhao Y, Huang XP, Zhu D, Wei M, Luo JC, Yu SY, et al. Deubiquitinase OTUD6A promotes breast cancer progression by increasing TopBP1 stability and rendering tumor cells resistant to DNA-damaging therapy. Cell Death Differ. 2022;29:2531–44.

    Article  CAS  PubMed  Google Scholar 

  18. Yao D, Dong M, Dai C, Wu S. Inflammation and inflammatory cytokine contribute to the initiation and development of ulcerative colitis and its associated cancer. Inflamm Bowel Dis. 2019;25:1595–602.

    Article  PubMed  Google Scholar 

  19. Zhen Y, Zhang H. NLRP3 Inflammasome and inflammatory bowel disease. Front Immunol. 2019;10:276.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Zhou LL, Liu T, Huang B, Luo M, Chen ZH, Zhao ZY, et al. Excessive deubiquitination of NLRP3-R779C variant contributes to very-early-onset inflammatory bowel disease development. J Allergy Clin Immunol. 2021;147:267–79.

    Article  CAS  PubMed  Google Scholar 

  21. Py BF, Kim MS, Vakifahmetoglu-Norberg H, Yuan JY. Deubiquitination of NLRP3 by BRCC3 Critically Regulates Inflammasome Activity. Mol Cell. 2013;49:331–8.

    Article  CAS  PubMed  Google Scholar 

  22. Ren GM, Zhang XY, Xiao Y, Zhang W, Wang Y, Ma WB, et al. ABRO1 promotes NLRP3 inflammasome activation through regulation of NLRP3 deubiquitination. Embo J. 2019;38:e100376.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Palazon-Riquelme P, Worboys JD, Green J, Valera A, Martin-Sanchez F, Pellegrini C, et al. USP7 and USP47 deubiquitinases regulate NLRP3 inflammasome activation. Embo Rep. 2018;19:e44766.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Liu T, Wang LQ, Liang PP, Wang XJ, Liu YK, Cai J, et al. USP19 suppresses inflammation and promotes M2-like macrophage polarization by manipulating NLRP3 function via autophagy. Cell Mol Immunol. 2021;18:2431–42.

    Article  CAS  PubMed  Google Scholar 

  25. Wang D, Zhang YN, Xu XM, Wu JF, Peng Y, Li J, et al. YAP promotes the activation of NLRP3 inflammasome via blocking K27-linked polyubiquitination of NLRP3. Nat Commun. 2021;12:2674.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Song H, Liu BY, Huai WW, Yu ZX, Wang WW, Zhao J, et al. The E3 ubiquitin ligase TRIM31 attenuates NLRP3 inflammasome activation by promoting proteasomal degradation of NLRP3. Nat Commun. 2016;7:13727.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Yan CY, Ouyang SH, Wang X, Wu YP, Sun WY, Duan WJ, et al. Celastrol ameliorates Propionibacterium acnes/LPS-induced liver damage and MSU-induced gouty arthritis via inhibiting K63 deubiquitination of NLRP3. Phytomedicine. 2021;80:153398.

    Article  CAS  PubMed  Google Scholar 

  28. Pu T, Liu WZ, Wu YJ, Zhao Y. A20 functions as a negative regulator in macrophage for DSS-induced colitis. Int Immunopharmacol. 2021;97:107804.

    Article  CAS  PubMed  Google Scholar 

  29. Zhou MX, He J, Shi YY, Liu XM, Luo SJ, Cheng C, et al. ABIN3 negatively regulates necroptosis-induced intestinal inflammation through recruiting A20 and restricting the ubiquitination of RIPK3 in inflammatory bowel disease. J Crohns Colitis. 2021;15:99–114.

    Article  PubMed  Google Scholar 

  30. Leszczynska K, Jakubczyk D, Gorska S. The NLRP3 inflammasome as a new target in respiratory disorders treatment. Front Immunol. 2022;13:1006654.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Zheng Y, Xu L, Dong N, Li F. NLRP3 inflammasome: The rising star in cardiovascular diseases. Front Cardiovasc Med. 2022;9:927061.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Khot M, Sood A, Tryphena KP, Khan S, Srivastava S, Singh SB, et al. NLRP3 inflammasomes: a potential target to improve mitochondrial biogenesis in Parkinson’s disease. Eur J Pharmacol. 2022;934:175300.

    Article  CAS  PubMed  Google Scholar 

  33. Cui S, Wang CH, Bai WZ, Li J, Pan Y, Huang XY, et al. CD1d1 intrinsic signaling in macrophages controls NLRP3 inflammasome expression during inflammation. Sci Adv. 2020;6:eaaz7290.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Song Y, Zhao Y, Ma Y, Wang Z, Rong L, Wang B, et al. Biological functions of NLRP3 inflammasome: A therapeutic target in inflammatory bowel disease. Cytokine Growth factor Rev. 2021;60:61–75.

    Article  CAS  PubMed  Google Scholar 

  35. Bauer C, Duewell P, Mayer C, Lehr HA, Fitzgerald KA, Dauer M, et al. Colitis induced in mice with dextran sulfate sodium (DSS) is mediated by the NLRP3 inflammasome. Gut. 2010;59:1192–9.

    Article  CAS  PubMed  Google Scholar 

  36. Li H, Guan Y, Liang B, Ding P, Hou X, Wei W, et al. Therapeutic potential of MCC950, a specific inhibitor of NLRP3 inflammasome. Eur J Pharmacol. 2022;928:175091.

    Article  CAS  PubMed  Google Scholar 

  37. Juliana C, Fernandes-Alnemri T, Kang S, Farias A, Qin F, Alnemri ES. Non-transcriptional priming and deubiquitination regulate NLRP3 inflammasome activation. J Biol Chem. 2012;287:36617–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Yu P, Zhang X, Liu N, Tang L, Peng C, Chen X. Pyroptosis: mechanisms and diseases. Signal Transduct Target Ther. 2021;6:128.

    Article  PubMed  PubMed Central  Google Scholar 

  39. Xu T, Yu WW, Fang H, Wang Z, Chi ZX, Guo XC, et al. Ubiquitination of NLRP3 by gp78/Insig-1 restrains NLRP3 inflammasome activation. Cell Death Differ. 2022;29:1582–95.

    Article  CAS  PubMed  Google Scholar 

  40. Song H, Zhao CY, Yu ZX, Li QZ, Yan RZ, Qin Y, et al. UAF1 deubiquitinase complexes facilitate NLRP3 inflammasome activation by promoting NLRP3 expression. Nat Commun. 2020;11:6042.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Tang J, Tu S, Lin GX, Guo H, Yan CK, Liu QJ, et al. Sequential ubiquitination of NLRP3 by RNF125 and Cbl-b limits inflammasome activation and endotoxemia. J Exp Med. 2020;217:e20182091.

    Article  PubMed  PubMed Central  Google Scholar 

  42. Liu X, Luo W, Chen J, Hu C, Mutsinze RN, Wang X, et al. USP25 deficiency exacerbates acute pancreatitis via up-regulating TBK1-NF-kappaB signaling in macrophages. Cell Mol Gastroenterol Hepatol. 2022;14:1103–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Wirtz S, Neufert C, Weigmann B, Neurath MF. Chemically induced mouse models of intestinal inflammation. Nat Protoc. 2007;2:541–6.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We graciously thank Fuping You (Peking University, Beijing, China) for sharing Otud6a−/− mice. This study was supported by the National Natural Science Foundation of China (81970338 and 82170373 to YW), Medical and Health Research Project of Zhejiang Province (2023XY164 to LH), and Key Scientific Project of Wenzhou City (ZY2021021 to YW).

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Authors and Affiliations

  1. Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang, China

    Xin Liu, Yi Fang, Chenghong Hu, Lingxi Zhang, Bo Jin, Wu Luo & Yi Wang

  2. Affiliated Yongkang First People’s Hospital and School of Pharmacy, Hangzhou Medical College, Hangzhou, Zhejiang, China

    Xin Liu, Xinting Lv & Guang Liang

  3. Department of Pathology, the Affiliated Quzhou Hospital of Wenzhou Medical University, Quzhou, Zhejiang, China

    Guorong Chen

  4. Department of Gastroenterology, the Affiliated Xiangshan Hospital of Wenzhou Medical University, Ningbo, Zhejiang, China

    Lijiang Huang & Yi Wang

  5. School of Pharmacy, Hangzhou Normal University, Hangzhou, Zhejiang, China

    Yi Wang

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Contributions

XLiu, GL and YW contributed to the literature search and study design. XLiu, YF, XLv, CH, LZ and BJ performed the experiments and analyzed the data. GC, LH and WL provided technical help. XLiu, GL and YW participated in the drafting of the article. All authors agree to be accountable for all aspects of work ensuring integrity and accuracy.

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Correspondence to Yi Wang.

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The authors declare no competing interests.

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The study about the human colonic biopsy samples was approved by the Human Ethical Committee of the Quzhou People’s Hospital (Approval document #2022–0037). All animal studies were approved by the Institutional Animal Policy and Welfare Committee of Wenzhou Medical University (Approval document #wydw2022-0202).

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Liu, X., Fang, Y., Lv, X. et al. Deubiquitinase OTUD6A in macrophages promotes intestinal inflammation and colitis via deubiquitination of NLRP3. Cell Death Differ (2023). https://doi.org/10.1038/s41418-023-01148-7

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