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Engraftment of connexin 43-expressing cells prevents post-infarct arrhythmia

Nature volume 450, pages 819–824 (2007)Cite this article

Abstract

Ventricular tachyarrhythmias are the main cause of sudden death in patients after myocardial infarction. Here we show that transplantation of embryonic cardiomyocytes (eCMs) in myocardial infarcts protects against the induction of ventricular tachycardia (VT) in mice. Engraftment of eCMs, but not skeletal myoblasts (SMs), bone marrow cells or cardiac myofibroblasts, markedly decreased the incidence of VT induced by in vivo pacing. eCM engraftment results in improved electrical coupling between the surrounding myocardium and the infarct region, and Ca2+ signals from engrafted eCMs expressing a genetically encoded Ca2+ indicator could be entrained during sinoatrial cardiac activation in vivo. eCM grafts also increased conduction velocity and decreased the incidence of conduction block within the infarct. VT protection is critically dependent on expression of the gap-junction protein connexin 43 (Cx43; also known as Gja1): SMs genetically engineered to express Cx43 conferred a similar protection to that of eCMs against induced VT. Thus, engraftment of Cx43-expressing myocytes has the potential to reduce life-threatening post-infarct arrhythmias through the augmentation of intercellular coupling, suggesting autologous strategies for cardiac cell-based therapy.

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Figure 1: VT in control, sham-injected and SM-engrafted hearts.
Figure 2: VT protection in eCM-engrafted hearts.
Figure 3: Entrainment of engrafted eCMs in vivo.
Figure 4: eCM engraftment improves conduction and prevents re-entry in Langendorff-perfused hearts.
Figure 5: VT protection in Cx43-expressing SM-engrafted hearts.

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Acknowledgements

We thank D. Fuerst for providing the anti-nebulin antibody; L. Field for providing the transgenic α-MHC–EGFP mouse line; W. Bloch for advice on immunostaining; C. Schaffer and N. Nishimura for technical advice; R. Gilmour Jr and N. Otani for comments on the manuscript; C. Fuegemann for the preparation of cardiac myofibroblasts; and H. Begerau, M. Czechowski, B. Eixmann, F. Holst, H. Doerr, K. Granitza and C. Russell for technical help. This study was supported by grants from the Deutsche Forschungsgemeinschaft (to W.R. and B.K.F.), the Federal Ministry of Education and Research, Germany (to T.L.), the European Commission (to B.K.F.), BONFOR (to J.W.S., A.H.) and the National Institutes of Health (to Y.N.T., M.I.K. and G.S.).

Author Contributions W.R., T.L. and P.S. contributed equally to this work. W.R. performed microsurgery, left-ventricular catheterization and analysis of data. T.L. supervised and analysed in vivo electrophysiology. P.S. performed in vivo imaging experiments, analysis and immunohistochemistry. Y.N.T. was involved in the in vivo imaging experiments and immunohistochemistry. B.-R.C. was involved in optical imaging and analysis of data. M.B. was involved in morphometry, immunohistochemistry and establishment of cardiac fibroblasts. R.D. was involved in the analysis of in vivo imaging experiments. U.B. was involved in the preparation of skeletal myoblasts, and in morphometry and immunohistochemistry. S.-M.H. was involved in the optical-imaging experiments and analysis of data. T.B. was involved in microsurgery, morphometry, left ventricular catheterization and immunohistochemistry. J.V.M. was involved in mouse breeding, immunohistochemistry and western blotting of tissues from the Cx43-expressing mouse. A.H. was involved in the generation of the lentivirus constructs. S.R. was involved in mouse breeding and in vivo imaging experiments. B.D. was involved in generation of the Cx43-expressing mouse model. B.G. was involved in Langendorff perfusion and optical-imaging experiments. A.P. supervised lentiviral work. A.W. supervised the microsurgery. G.S. was involved in the optical-imaging experiments, their analysis and writing of the manuscript. J.W.S. performed electrophysiological experiments in vivo and analysed data. M.I.K. designed experiments, analysed in vivo imaging experiments and wrote the manuscript. B.K.F. initiated the study, designed experiments and wrote the manuscript.

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  1. Wilhelm Roell, Thorsten Lewalter and Philipp Sasse: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Physiology I, Life and Brain Center,,

    Wilhelm Roell, Philipp Sasse, Martin Breitbach, Ulrich M. Becher, Toktam Bostani, Jan W. Schrickel & Bernd K. Fleischmann

  2. Department of Cardiac Surgery,,

    Wilhelm Roell, Toktam Bostani & Armin Welz

  3. Department of Internal Medicine II,,

    Thorsten Lewalter, Ulrich M. Becher & Jan W. Schrickel

  4. Institute of Genetics,,

    Julia von Maltzahn, Britta Eiberger & Klaus Willecke

  5. Institute of Pharmacology, University of Bonn, Bonn 53105, Germany ,

    Andreas Hofmann & Alexander Pfeifer

  6. Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853-6401, USA,

    Yvonne N. Tallini, Robert Doran, Shaun Reining & Michael I. Kotlikoff

  7. Cardiovascular Research Center, Rhode Island Hospital and Brown Medical School, Providence, Rhode Island 02903, USA ,

    Bum-Rak Choi

  8. Department of Cell Biology and Physiology, University of Pittsburgh, School of Medicine, Pittsburgh, Pennsylvania 15261, USA,

    Seong-Min Hwang, Bethann Gabris & Guy Salama

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Correspondence to Michael I. Kotlikoff or Bernd K. Fleischmann.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-4 with Legends, Supplementary Table 1 and Supplementary Methods with references (PDF 5367 kb)

Supplementary Video 1

This file contains Supplementary Video 1 which shows in vivo monitoring of functional engraftment of GCaMP2+ embryonic cardiomyocytes in the infarct. Ca2+ signaling is detected within a section of infarcted left ventricle during sinus rhythm (heart rate: 490 bpm) in an anesthetized and ventilated mouse with open chest. Engrafted cells are coupled (2:1) to the native myocardium. GCaMP2 fluorescence only increases in case of coupling, the paradox movement of the infarcted area does not alter the fluorescence intensity. Recorded at 112 fps and displayed at 11.2 fps. (MOV 2570 kb)

Supplementary Video 2

This file contains Supplementary Video 2 which shows optical voltage recordings in infarcted, sham injected mouse. Di-4-ANEPPS fluorescence signals during S1-S2 stimulation protocol in an isolated heart. The first 2 waves are produced by stimulations at 150 ms, followed by early activation at 80 ms cycle length (white dot indicates S2 start). Note the movement of the voltage wave around the infarct area and the induction of prominent circular waves by S2 stimulation (see also Fig. 4d). Recorded at 1 kHz and displayed at 66 fps. (MOV 3690 kb)

Supplementary Video 3

This file contains Supplementary Video 3 which shows that engraftment of embryonic cardiomyocytes results in conduction of voltage wave into the infarct. Same stimulation protocol as in Supplementary Video 2. Note partial invasion of infarct by voltage wave during S2 stimulations and failure of S2 protocol (white dot) to produce boundary effects. Engraftment was confirmed by visualization of fluorescence of engrafted cells. Recorded at 1 kHz and displayed at 66 fps. (MOV 3448 kb)

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Roell, W., Lewalter, T., Sasse, P. et al. Engraftment of connexin 43-expressing cells prevents post-infarct arrhythmia. Nature 450, 819–824 (2007). https://doi.org/10.1038/nature06321

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