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Methylation determines fibroblast activation and fibrogenesis in the kidney

Nature Medicine volume 16, pages 544–550 (2010)Cite this article

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Abstract

Fibrogenesis is a pathological wound repair process that fails to cease, even when the initial insult has been removed. Fibroblasts are principal mediators of fibrosis, and fibroblasts from fibrotic tissues fail to return to their quiescent stage, including when cultured in vitro. In a search for underlying molecular mechanisms, we hypothesized that this perpetuation of fibrogenesis is caused by epigenetic modifications. We demonstrate here that hypermethylation of RASAL1, encoding an inhibitor of the Ras oncoprotein, is associated with the perpetuation of fibroblast activation and fibrogenesis in the kidney. RASAL1 hypermethylation is mediated by the methyltransferase Dnmt1 in renal fibrogenesis, and kidney fibrosis is ameliorated in Dnmt1+/− heterozygous mice. These studies demonstrate that epigenetic modifications may provide a molecular basis for perpetuated fibroblast activation and fibrogenesis in the kidney.

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Figure 1: 5′-azacytidine ameliorates experimental renal fibrosis.
Figure 2: RASAL1 hypermethylation in fibrotic kidney fibroblasts.
Figure 3: Absence of Rasal1 hypermethylation in kidneys that do not become fibrotic upon injury.
Figure 4: Rasal1 silencing causes renal fibroblast activation via Ras hyperactivity.
Figure 5: Dnmts in the mouse model of folic acid–induced nephropathy.
Figure 6: TGF-β1–induced methylation of Rasal1 in kidney fibroblasts is mediated by Dnmt1.

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Acknowledgements

This study was partially funded by research grants R01DK081576 (M.Z.), R03DK081687 (M.Z.), R01DK30932 (D.J.S.), R01DK55001 (R.K.), R01CA125550 (R.K.), Mentored Clinical Scientist Development Awards 1K08 CA129204 (E.M.Z.) and K08 DK074558 (M.Z.) from the National Institutes of Health, the American Society of Nephrology Carl W. Gottschalk Scholar Grant (M.Z.), American Heart Association Scientist Development Grant SDG0735602T, Deutsche Forschungsgemeinschaft-Stipendium BE4211/1-1 (W.B.), Champalimaud Metastasis Research Program (R.K.) and research funds from the Beth Israel Deaconess Medical Center for the Division of Matrix Biology. We thank A. Lau and T. Zhong Hu for technical assistance. FSP-1–specific antibodies were a gift from E. Neilson (Vanderbilt University).

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Authors and Affiliations

  1. Division of Matrix Biology, Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts, USA

    Wibke Bechtel, Scott McGoohan, Elisabeth M Zeisberg, Raghu Kalluri & Michael Zeisberg

  2. Department of Nephrology and Rheumatology, Georg-August-University Medical Center, Göttingen, Germany

    Gerhard A Müller

  3. Section for Transplantation Immunology and Immunohematology, Center for Medical Research, University Medical Clinic, Tübingen, Germany

    Hubert Kalbacher & Claudia A Müller

  4. Renal Section, Boston University Medical Center, Boston, Massachusetts, USA

    David J Salant

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Contributions

W.B. performed and designed experiments, analyzed data and edited the manuscript. S.M. performed experiments and analyzed data. E.M.Z. advised, performed experiments, analyzed data and edited the manuscript. G.A.M. and C.A.M. characterized and provided human fibroblasts. H.K. generated Rasal1 antibodies. D.J.S. provided nephrotoxic serum and edited the manuscript. R.K. advised and edited the manuscript. M.Z. designed, performed and supervised experiments, analyzed data and wrote the manuscript.

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Correspondence to Michael Zeisberg.

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Bechtel, W., McGoohan, S., Zeisberg, E. et al. Methylation determines fibroblast activation and fibrogenesis in the kidney. Nat Med 16, 544–550 (2010). https://doi.org/10.1038/nm.2135

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