The AMPK–Parkin axis negatively regulates necroptosis and tumorigenesis by inhibiting the necrosome

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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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Fig. 1: Parkin KO mice have increased inflammation and spontaneous tumour formation.
Fig. 2: Parkin negatively regulates RIPK1−RIPK3 interaction.
Fig. 3: Parkin promotes K33-linked polyubiquitination of RIPK3.
Fig. 4: Parkin-mediated RIPK3 ubiquitination is important for RIPK3 inhibition.
Fig. 5: Parkin is activated by AMPK-dependent phosphorylation at S9 during necroptosis.
Fig. 6: RIPK3 inhibition suppresses intestinal inflammation and colitis-associated tumorigenesis in an AOM−DSS mouse model.
Fig. 7: AMPK activation inhibits colitis-associated cancer in a Parkin-dependent manner.

Data availability

Source data for Figs. 1b–f, 2d,h,j, 3g, 4a,c,g, 5a,j, 6b,c,e,h–j and 7b–d,f and Supplementary Figs. 1a,b,e,f, 4e–g, 5d, 6c–f and 7a,b,d–f,h,i are provided in Supplementary Table 1. All data supporting the findings of this study are available from the corresponding author on reasonable request.

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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.

Author information

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.

Correspondence to Jin-San Zhang or Zhenkun Lou.

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Supplementary information

Supplementary Information

Supplementary Figures 1−8 and Supplementary table titles/legends.

Reporting Summary

Supplementary Table 1

Statistics source data

Supplementary Table 2

Antibodies

Supplementary Table 3

Parkin and PINK1 shRNAs

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