Abstract
The receptor-interacting serine/threonine-protein kinases RIPK1 and RIPK3 play important roles in necroptosis that are closely linked to the inflammatory response. Although the activation of necroptosis is well characterized, the mechanism that tunes down necroptosis is largely unknown. Here we find that Parkin (also known as PARK2), an E3 ubiquitin ligase implicated in Parkinson’s disease and as a tumour suppressor, regulates necroptosis and inflammation by regulating necrosome formation. Parkin prevents the formation of the RIPK1−RIPK3 complex by promoting polyubiquitination of RIPK3. Parkin is phosphorylated and activated by the cellular energy sensor AMP-activated protein kinase (AMPK). Parkin deficiency potentiates the RIPK1−RIPK3 interaction, RIPK3 phosphorylation and necroptosis. Parkin deficiency enhances inflammation and inflammation-associated tumorigenesis. These findings demonstrate that the AMPK−Parkin axis negatively regulates necroptosis by inhibiting RIPK1−RIPK3 complex formation; this regulation may serve as an important mechanism to fine-tune necroptosis and inflammation.
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References
Long, J. S. & Ryan, K. M. New frontiers in promoting tumour cell death: targeting apoptosis, necroptosis and autophagy. Oncogene 31, 5045–5060 (2012).
Su, Z., Yang, Z., Xu, Y., Chen, Y. & Yu, Q. Apoptosis, autophagy, necroptosis, and cancer metastasis. Mol. Cancer 14, 48 (2015).
Lalaoui, N., Lindqvist, L. M., Sandow, J. J. & Ekert, P. G. The molecular relationships between apoptosis, autophagy and necroptosis. Semin. Cell Dev. Biol. 39, 63–69 (2015).
Radogna, F., Dicato, M. & Diederich, M. Cancer-type-specific crosstalk between autophagy, necroptosis and apoptosis as a pharmacological target. Biochem. Pharmacol. 94, 1–11 (2015).
Tanaka, T. et al. A novel inflammation-related mouse colon carcinogenesis model induced by azoxymethane and dextran sodium sulfate. Cancer Sci. 94, 965–973 (2003).
Neufert, C., Becker, C. & Neurath, M. F. An inducible mouse model of colon carcinogenesis for the analysis of sporadic and inflammation-driven tumor progression. Nat. Protoc. 2, 1998–2004 (2007).
Welz, P. S. et al. FADD prevents RIP3-mediated epithelial cell necrosis and chronic intestinal inflammation. Nature 477, 330–334 (2011).
Rubin, D. C., Shaker, A. & Levin, M. S. Chronic intestinal inflammation: inflammatory bowel disease and colitis-associated colon cancer. Front. Immunol. 3, 107 (2012).
Grivennikov, S. I. Inflammation and colorectal cancer: colitis-associated neoplasia. Semin. Immunopathol. 35, 229–244 (2013).
Kim, E. R. & Chang, D. K. Colorectal cancer in inflammatory bowel disease: the risk, pathogenesis, prevention and diagnosis. World J. Gastroenterol. 20, 9872–9881 (2014).
Pasparakis, M. & Vandenabeele, P. Necroptosis and its role in inflammation. Nature 517, 311–320 (2015).
Seifert, L. et al. The necrosome promotes pancreatic oncogenesis via CXCL1 and Mincle-induced immune suppression. Nature 532, 245–249 (2016).
Cho, Y. S. et al. Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation. Cell 137, 1112–1123 (2009).
Sun, L. et al. Mixed lineage kinase domain-like protein mediates necrosis signaling downstream of RIP3 kinase. Cell 148, 213–227 (2012).
Wu, X. N. et al. Distinct roles of RIP1−RIP3 hetero- and RIP3−RIP3 homo-interaction in mediating necroptosis. Cell Death Differ. 21, 1709–1720 (2014).
Davies, S. P., Carling, D. & Hardie, D. G. Tissue distribution of the AMP-activated protein kinase, and lack of activation by cyclic-AMP-dependent protein kinase, studied using a specific and sensitive peptide assay. Eur. J. Biochem. 186, 123–128 (1989).
Kemp, B. E. et al. AMP-activated protein kinase, super metabolic regulator. Biochem. Soc. Trans. 31, 162–168 (2003).
Shimura, H. et al. Familial Parkinson disease gene product, parkin, is a ubiquitin-protein ligase. Nat. Genet. 25, 302–305 (2000).
Yamamoto, A. et al. Parkin phosphorylation and modulation of its E3 ubiquitin ligase activity. J. Biol. Chem. 280, 3390–3399 (2005).
Moore, D. J., West, A. B., Dikeman, D. A., Dawson, V. L. & Dawson, T. M. Parkin mediates the degradation-independent ubiquitination of Hsp70. J. Neurochem. 105, 1806–1819 (2008).
Chen, D. et al. Parkin mono-ubiquitinates Bcl-2 and regulates autophagy. J. Biol. Chem. 285, 38214–38223 (2010).
Iguchi, M. et al. Parkin-catalyzed ubiquitin-ester transfer is triggered by PINK1-dependent phosphorylation. J. Biol .Chem. 288, 22019–22032 (2013).
Kane, L. A. et al. PINK1 phosphorylates ubiquitin to activate Parkin E3 ubiquitin ligase activity. J. Cell Biol. 205, 143–153 (2014).
Koyano, F. et al. Ubiquitin is phosphorylated by PINK1 to activate parkin. Nature 510, 162–166 (2014).
Picchio, M. C. et al. Alterations of the tumor suppressor gene Parkin in non-small cell lung cancer. Clin. Cancer Res. 10, 2720–2724 (2004).
Fujiwara, M. et al. Parkin as a tumor suppressor gene for hepatocellular carcinoma. Oncogene 27, 6002–6011 (2008).
Poulogiannis, G. et al. PARK2 deletions occur frequently in sporadic colorectal cancer and accelerate adenoma development in Apc mutant mice. Proc. Natl Acad. Sci. USA 107, 15145–15150 (2010).
Veeriah, S. et al. Somatic mutations of the Parkinson’s disease-associated gene PARK2 in glioblastoma and other human malignancies. Nat. Genet. 42, 77–82 (2010).
Lee, S. B. et al. Parkin regulates mitosis and genomic stability through Cdc20/Cdh1. Mol. Cell 60, 21–34 (2015).
Lee, S. et al. Multiple-level validation identifies PARK2 in the development of lung cancer and chronic obstructive pulmonary disease. Oncotarget 7, 44211–44223 (2016).
Thomas, M. Inflammation in Parkinson’s Disease (Springer, 2014).
Noble, C. L. et al. Regional variation in gene expression in the healthy colon is dysregulated in ulcerative colitis. Gut 57, 1398–1405 (2008).
Li, J. et al. The RIP1/RIP3 necrosome forms a functional amyloid signaling complex required for programmed necrosis. Cell 150, 339–350 (2012).
Deng, M. et al. Deubiquitination and activation of AMPK by USP10. Mol. Cell 61, 614–624 (2016).
Gonzalez-Juarbe, N. et al. Pore-forming toxins induce macrophage necroptosis during acute bacterial pneumonia. PLoS Pathog. 11, e1005337 (2015).
Wagener, C., Stocking, C. & Müller, O. Cancer Signaling: From Molecular Biology to Targeted Therapy (Wiley-Blackwell, 2016).
Gwinn, D. M. et al. AMPK phosphorylation of raptor mediates a metabolic checkpoint. Mol. Cell 30, 214–226 (2008).
Canto, C. et al. AMPK regulates energy expenditure by modulating NAD+ metabolism and SIRT1 activity. Nature 458, 1056–1060 (2009).
Mandal, P. et al. RIP3 induces apoptosis independent of pronecrotic kinase activity. Mol. Cell 56, 481–495 (2014).
Yang, X. S. et al. Hypoxia-inducible factor-1 alpha is involved in RIP-induced necroptosis caused by in vitro and in vivo ischemic brain injury. Sci. Rep. 7, 5818 (2017).
Los, M. et al. Activation and caspase-mediated inhibition of PARP: a molecular switch between fibroblast necrosis and apoptosis in death receptor signaling. Mol. Biol. Cell 13, 978–988 (2002).
Eguchi, Y., Shimizu, S. & Tsujimoto, Y. Intracellular ATP levels determine cell death fate by apoptosis or necrosis. Cancer Res. 57, 1835–1840 (1997).
Tsujimoto, Y. Apoptosis and necrosis: intracellular ATP level as a determinant for cell death modes. Cell Death Differ. 4, 429–434 (1997).
Nikoletopoulou, V., Markaki, M., Palikaras, K. & Tavernarakis, N. Crosstalk between apoptosis, necrosis and autophagy. Biochim. Biophys. Acta 1833, 3448–3459 (2013).
Rothfuss, O. et al. Parkin protects mitochondrial genome integrity and supports mitochondrial DNA repair. Hum. Mol. Genet. 18, 3832–3850 (2009).
Omoto, S. et al. Suppression of RIPK3-dependent necroptosis by human cytomegalovirus. J. Biol. Chem. 290, 11635–11648 (2015).
Acknowledgements
We thank all members of the Lou lab for their critical discussion of this work. This work was supported by NIH grant numbers CA203561 and CA224921 to Z.L. and was also supported by grant number ENP-RES20180401-02 to S.B.L.
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S.B.L. and J.J.K. designed and performed most of the experiments, analysed data and had a lead author role in the preparation of the manuscript. S.B.L., J.J.K., A.A., K.A. and P.Y. performed the animal experiments. F.C.F. and W.S. provided the S65-ubiquitin homemade antibody. S.-A.H, Y.F., S.S.K., S.-Y.P., Q.L., J.O.C., S.I.C., S.N., A.A., K.A. and S.-Y.T. helped with the immunohistochemical analysis of human colon and inflammation-related small bowel tissue. S.N. participated in the proofreading of this manuscript. L.-S.G. provided reagents for experiments. J.-S.Z. participated in the data analysis and manuscript writing, and provided technical assistance and materials. Z.L. conceived and supervised the project.
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Parkin and PINK1 shRNAs
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Lee, S.B., Kim, J.J., Han, SA. et al. The AMPK–Parkin axis negatively regulates necroptosis and tumorigenesis by inhibiting the necrosome. Nat Cell Biol 21, 940–951 (2019). https://doi.org/10.1038/s41556-019-0356-8
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DOI: https://doi.org/10.1038/s41556-019-0356-8
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