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Tumor necrosis factor-α confers cardioprotection through ectopic expression of keratins K8 and K18

Abstract

Tumor necrosis factor-α (TNF-α), one of the major stress-induced proinflammatory cytokines, is upregulated in the heart after tissue injury1,2, and its sustained expression can contribute to the development of heart failure1,3,4. Whether TNF-α also exerts cytoprotective effects in heart failure is not known. Here we provide evidence for a cardioprotective function of TNF-α in a genetic heart failure model, desmin-deficient mice. The cardioprotective effects of TNF-α are a consequence of nuclear factor-κB (NF-κB)-mediated ectopic expression in cardiomyocytes of keratin 8 (K8) and keratin 18 (K18), two epithelial-specific intermediate filament proteins5,6. In cardiomyocytes, K8 and K18 (K8/K18) formed an alternative cytoskeletal network that localized mainly at intercalated discs (IDs) and conferred cardioprotection by maintaining normal ID structure and mitochondrial integrity and function. Ectopic induction of K8/K18 expression in cardiomyocytes also occurred in other genetic and experimental models of heart failure. Loss of the K8/K18 network resulted in a maladaptive cardiac phenotype following transverse aortic constriction. In human failing myocardium, where TNF-α expression is upregulated2, K8/K18 were also ectopically expressed and localized primarily at IDs, which did not contain detectable amounts of desmin. Thus, TNF-α– and NF-κB-mediated formation of an alternative, stress-induced intermediate filament cytoskeleton has cardioprotective function in mice and potentially in humans.

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Figure 1: TNF-α overexpression rescues myocardial degeneration, ultrastructural abnormalities, mitochondrial defects and cardiac dysfunction caused by desmin deficiency.
Figure 2: K8 and K18 are ectopically expressed in TNF-α–overexpressing cardiomyocytes.
Figure 3: Ectopic expression of K8/K18 confers cardioprotection in TnfMyh6;Des−/− mice through maintenance of mitochondrial structure and function and of intercalated disc integrity.
Figure 4: Ectopic induction of K8 and K18 in cardiomyocytes is a common response in several genetic and experimental cardiomyopathy models, as well as in human heart failure.

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Acknowledgements

We thank W. Franke and Y. Dörflinger (Helmholtz Group for Cell Biology, German Cancer Research Center, Heidelberg) for extensive technical support with EM and immunogold labeling, for providing EM-related materials, antibodies and human tissue samples, and for very valuable comments. We especially thank L. Scorrano (University of Padova) for his help and support on the study of mitochondrial function and for providing the consumables for mitochondrial function experiments for these experiments. We thank L. Staloch, C. Weinheimer and A. Kovachs for the TAC surgeries and echocarardiographic measurements of the TACed mice. We also thank W.K. Jones (Department of Pharmacology and Cell Biophysics, University of Cincinnati) for kindly providing the IκBα-3M mice, M. Gerstenlauer for technical assistance with IKKMyHC mice experiments and J. D. Molkentin (Howard Hughes Medical Institute and Cincinnati Children's Hospital Medical Center) for providing samples of MHC-Cn and Csrp3−/− mice and mice subjected to TAC or MI. We also thank P. Politis and N. Athanassiadis (Biomedical Research Foundation, Academy of Athens) for providing materials related to the luciferase reporter assays and human samples, respectively. Our sincere thanks to I. Kostavasili for her long-term technical assistance throughout this project. This work was supported by the Greek Secretariat of Research and Development grants (PENED 01ED371, PEP-ATT-39, ESPA SYNERGASIA 09SYN-21-965) and grant of “Excellence II” 5342 to Y.C., a US National Institutes of Health R01 HL 111094 grant to D.L.M. and a fellowship (Heracleitus II) by the European Union and Greek national funds through the Operational Program “Education and Lifelong Learning” of the National Strategic Reference Framework to S.P.

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Authors

Contributions

Y.C. and S.P. conceived the project, designed research and analyzed all the data. S.P. performed the research. S.R. helped with the EM experiments. M.E.S. helped with the mitochondrial functional experiments. T.G.S. helped with the experiments on several models of heart failure. C.H.D. and A.V. helped with echocardiography. H.J.M. contributed with analyses of the IKKMyHC mice. L.K. provided the human samples. D.L.M. provided the original TnfMyh6 mice, helped with TAC experiments and data interpretation. S.P. and Y.C. wrote the paper, and Y.C. supervised all aspects of the work. All co-authors approved the submitted manuscript.

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Correspondence to Yassemi Capetanaki.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–12 & Supplementary Tables 1–3 (PDF 27007 kb)

Dataset 1

GO analysis of the mostly altered genes in TnfMyh6Des−/− myocardium. (XLS 71 kb)

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Papathanasiou, S., Rickelt, S., Soriano, M. et al. Tumor necrosis factor-α confers cardioprotection through ectopic expression of keratins K8 and K18. Nat Med 21, 1076–1084 (2015). https://doi.org/10.1038/nm.3925

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