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Connective tissue growth factor: a crucial cytokine-mediating cardiac fibrosis in ongoing enterovirus myocarditis

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Abstract

Dilated cardiomyopathy (DCM) as a consequence of viral myocarditis is a worldwide cause of morbidity and death. The deposition of matrix proteins, such as collagen, in the course of ongoing viral myocarditis results in cardiac remodeling and finally in cardiac fibrosis, the hallmark of DCM. To identify mediators of virus-induced cardiac fibrosis, microarray analysis was conducted in a murine model of chronic coxsackievirus B3 (CVB3) myocarditis. By this attempt, we identified connective tissue growth factor (CTGF) as a novel factor highly expressed in infected hearts. Further investigations by quantitative reverse transcription polymerase chain reaction and Western blot analysis confirmed a strong induction of cardiac CTGF expression in the course of CVB3 myocarditis. By in situ hybridization and immunohistochemistry, basal CTGF messenger ribonucleic acid (mRNA) and protein expression were confined in noninfected control hearts mainly to endothelial cells, whereas in CVB3-infected hearts, also numerous fibroblasts were found to express CTGF. Regulation of CTGF is known to be basically mediated by transforming growth factor (TGF)-β. In the course of CVB3 myocarditis, CTGF upregulation coincided with increased cardiac TGF-β and procollagen type I mRNA expression, preceding the formation of fibrotic lesions. In in vitro experiments, we found that downregulation of CVB3 replication by means of small interfering RNAs (siRNAs) reverses the upregulation of CTGF mRNA expression. In contrast, downregulation of CTGF by siRNA molecules did not significantly reduce viral load, indicating that CTGF is not essential for CVB3 life cycle. The significantly enhanced transcript levels of TGF-β, CTGF, and procollagen type I in cultivated CVB3-infected primary cardiac fibroblasts substantiate the role of fibroblasts as a relevant cell population in cardiac remodeling processes. We conclude that CTGF is a crucial molecule in the development of fibrosis in ongoing enteroviral myocarditis. Thus, downregulation of cardiac CTGF expression may open novel therapeutic approaches counteracting the development of cardiac fibrosis and subsequent heart muscle dysfunction.

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Acknowledgments

This study was supported by the Deutsche Forschungsgemeinschaft, SFB/TR 19-04, the fortüne program of the Medical Faculty of Tübingen (1271-0), and in part by the “Fondazione Cassa di Risparmio of Trieste,” by the “Fondazione Benefica Katleen-Foreman Casali of Trieste.” G. Grassi is supported by the program “Rientro cervelli” art. 1 DM n.13, MIUR (Ministero dell’Istruzione, dell’Università e della Ricerca Scientifica.” The support of S. Berchtold, Institute for Medical Microbiology and of Sandra Bundschuh, Department of Molecular Pathology, University Hospital Tübingen, is acknowledged. There are no potential conflicts of interest anywhere, relating to this article.

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Authors and Affiliations

  1. Department of Molecular Pathology, Institute for Pathology, University Hospital Tübingen, Liebermeisterstr.8, 72076, Tübingen, Germany

    Christine Lang, Martina Sauter, Gudrun Szalay, Giorgia Racchi, Gabriele Grassi, Reinhard Kandolf & Karin Klingel

  2. Institute of Physiology, University of Tübingen, Tübingen, Germany

    Christine Lang & Florian Lang

  3. Department of Internal Medicine, University Hospital of Trieste, Cattinara, Strada di Fiume 447, 34149, Trieste, Italy

    Gabriele Grassi

  4. Istituto di Fisiologia Clinica, Area della Ricerca di Pisa, Pisa, Italy

    Giuseppe Rainaldi & Alberto Mercatanti

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  1. Christine Lang
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Correspondence to Karin Klingel.

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Lang, C., Sauter, M., Szalay, G. et al. Connective tissue growth factor: a crucial cytokine-mediating cardiac fibrosis in ongoing enterovirus myocarditis. J Mol Med 86, 49–60 (2008). https://doi.org/10.1007/s00109-007-0249-3

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  • DOI: https://doi.org/10.1007/s00109-007-0249-3

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