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CaMKII is a RIP3 substrate mediating ischemia- and oxidative stress–induced myocardial necroptosis

Nature Medicine volume 22, pages 175–182 (2016)Cite this article

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Abstract

Regulated necrosis (necroptosis) and apoptosis are crucially involved in severe cardiac pathological conditions, including myocardial infarction, ischemia-reperfusion injury and heart failure. Whereas apoptotic signaling is well defined, the mechanisms that underlie cardiomyocyte necroptosis remain elusive. Here we show that receptor-interacting protein 3 (RIP3) triggers myocardial necroptosis, in addition to apoptosis and inflammation, through activation of Ca2+-calmodulin–dependent protein kinase (CaMKII) rather than through the well-established RIP3 partners RIP1 and MLKL. In mice, RIP3 deficiency or CaMKII inhibition ameliorates myocardial necroptosis and heart failure induced by ischemia-reperfusion or by doxorubicin treatment. RIP3-induced activation of CaMKII, via phosphorylation or oxidation or both, triggers opening of the mitochondrial permeability transition pore and myocardial necroptosis. These findings identify CaMKII as a new RIP3 substrate and delineate a RIP3-CaMKII-mPTP myocardial necroptosis pathway, a promising target for the treatment of ischemia- and oxidative stress–induced myocardial damage and heart failure.

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Figure 1: RIP3 deficiency protects mouse hearts from I/R-induced necrosis and heart failure.
Figure 2: Dox-induced myocardial necrosis, cardiomyopathy and mortality are alleviated in Ripk3−/− mice.
Figure 3: RIP3-induced cardiomyocyte necroptosis is independent of RIP1 and MLKL.
Figure 4: RIP3-induced CaMKII activation is essential for cardiomyocyte necroptosis.
Figure 5: CaMKII is activated by RIP3 via both phosphorylation and oxidation.
Figure 6: RIP3-mediated cardiomyocyte apoptosis and necroptosis contribute to I/R-induced myocardial injury.

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Acknowledgements

We thank X.-D. Wang for Ripk3−/− mice, M. Anderson for CaMKII plasmids, and H. Cheng and I.C. Bruce for comments on the manuscript. This work was supported by the National Basic Research Program of China (2013CB531200 (Y.Z.), 2012CB518000 (R.-P.X.) and 2012CB944500 (C.-M.C.)), the National Natural Science Foundation of China (81170100 (Y.Z.), 81370234 (Y.Z.), 81130073 (R.-P.X.), 81370233 (C.-M.C.), 91439107 (C.-M.C.), 81270190 (F.L.) and 31221002 (R.-P.X.)), National Science and Technology Major Projects for Major New Drugs Innovation and Development (2013ZX09508104) (R.-P.X.) and the Specialized Research Fund for the Doctoral Program of Higher Education (20110001120119) (Y.Z.).

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  1. Ting Zhang and Yan Zhang: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Molecular Medicine, Peking University, Beijing, China

    Ting Zhang, Yan Zhang, Mingyao Cui, Li Jin, Yimei Wang, Fengxiang Lv, Yuli Liu, Wen Zheng, Haibao Shang, Jun Zhang, Mao Zhang, Hongkun Wu, Jiaojiao Guo, Xiuqin Zhang, Xinli Hu, Chun-Mei Cao & Rui-Ping Xiao

  2. State Key Laboratory of Biomembrane and Membrane Biotechnology, Peking University, Beijing, China

    Ting Zhang, Yan Zhang, Mingyao Cui, Li Jin, Yimei Wang, Fengxiang Lv, Yuli Liu, Wen Zheng, Haibao Shang, Jun Zhang, Mao Zhang, Hongkun Wu, Jiaojiao Guo, Xiuqin Zhang, Xinli Hu, Chun-Mei Cao & Rui-Ping Xiao

  3. Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China

    Rui-Ping Xiao

  4. Beijing City Key Laboratory of Cardiometabolic Molecular Medicine, Peking University, Beijing, China

    Rui-Ping Xiao

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Contributions

T.Z. and Y.Z. generated the initial idea and conducted key experiments. Y.Z. and R.-P.X. proposed the hypothesis, designed the study, supervised the experiments and data analysis, wrote the manuscript and interpreted results. T.Z., Y.Z., M.C., L.J., Y.W., F.L., Y.L., W.Z., H.S., J.Z., M.Z., H.W., J.G., X.Z., X.H. and C.-M.C. performed the experiments.

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Correspondence to Yan Zhang or Rui-Ping Xiao.

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Zhang, T., Zhang, Y., Cui, M. et al. CaMKII is a RIP3 substrate mediating ischemia- and oxidative stress–induced myocardial necroptosis. Nat Med 22, 175–182 (2016). https://doi.org/10.1038/nm.4017

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