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Secreted Frizzled-related protein 2 is a procollagen C proteinase enhancer with a role in fibrosis associated with myocardial infarction

Nature Cell Biology volume 11, pages 46–55 (2009)Cite this article

Abstract

Secreted Frizzled-related proteins (sFRPs) have emerged as key regulators of a wide range of developmental and disease processes. Most of the known functions of mammalian sFRPs have been attributed to their ability to antagonize Wnt signalling. Recently however, Xenopus laevis and zebrafish sFRP, Sizzled, was shown to function as an antagonist of Chordin processing by Tolloid-like metalloproteinases. This has led to the proposal that sFRPs may function as evolutionarily conserved antagonists of chordinase activities of this class of proteinases. In contrast to this proposal, we show here that the mammalian sFRP, sFRP2, does not affect Chordin processing, but instead, can serve as a direct enhancer of procollagen C proteinase activity of Tolloid-like metalloproteinases. We also show that the level of fibrosis, in which procollagen processing by Tolloid-like proteinases has a rate-limiting role, is markedly reduced in Sfrp2-null mice subjected to myocardial infarction. Importantly, this reduced level of fibrosis is accompanied by significantly improved cardiac function. This study thus uncovers a function for sFRP2 and a potential therapeutic application for sFRP2 antagonism in controlling fibrosis in the infarcted heart.

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Figure 1: sFRP2 enhances cleavage of procollagen, but not Chordin, by BMP1 and mTLL1.
Figure 2: sFRP2 binds BMP1 with a KD in the physiological range, predominantly through its Fzl domain.
Figure 3: sFRP2 and Szl bind Tolloid-like proteinases through non-protease domain sequences and BMP1/procollagen binding is enhanced by sFRP2.
Figure 4: Reduced processing of type I procollagen and deposition of collagen into ECM by Sfrp2−/− fibroblasts.
Figure 5: Induction of Sfrp2 and Bmp1 during the fibrosis phase in the infarcted heart.
Figure 6: Reduced fibrosis in the infarcted Sfrp2-deficient heart.
Figure 7: Improved cardiac function of the infarcted Sfrp2-null heart.

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Acknowledgements

We would like to thank Kuangfu Hsiao, Patricia Cobo and Guy Hoffman for technical assistance, and Tony Brown (Cornell) for providing information regarding qPCR primers for Wnt genes. K.K. and M.L. would like to acknowledge Rosemary Kraemer's teaching of technical procedures for the mouse myocardial infarction model. Work in the laboratory of A.K. is supported by funding from the German Research Foundation (DFG) for the Cluster of Excellence REBIRTH (from Regenerative Biology of Reconstructive Therapy), and by the European Union FP6 contract 'Heart Repair' (LSHM-CT-2005-018630). C.T.B. is an Established Investigator of the American Heart Association and is supported by the Snart Cardiovascular Fund. This work was supported in part by grants from the NIH (C.T.B, D.S.G, T.N.S).

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Author notes

  1. Koichi Kobayashi, Min Luo and Yue Zhang: These authors contributed equally to this work.

  2. Daniel S. Greenspan and Thomas N. Sato: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cell and Developmental Biology, Weill Medical College of Cornell University, New York, 1006S, NY, USA

    Koichi Kobayashi, Min Luo, David C. Wilkes, Chikaomi Yamada, Ting-Chun Liu, Craig T. Basson & Thomas N. Sato

  2. Greenberg Division of Cardiology and Department of Medicine, Weill Medical College of Cornell University, New York, 1006S, NY, USA

    David C. Wilkes & Craig T. Basson

  3. Program in Cellular and Molecular Biology, University of Wisconsin, Madison, 53706, Wisconsin, USA

    Yue Zhang & Daniel S. Greenspan

  4. the Departments of Pathology and Laboratory Medicine and Pharmacology, University of Wisconsin, Madison, Wisconsin 53706, USA.,

    Yue Zhang, Gaoxiang Ge, Guorui Huang & Daniel S. Greenspan

  5. Institut für Molekularbiologie, OE5250, Medizinische Hochschule Hannover, Hannover, D-30625, Germany

    Thomas Grieskamp & Andreas Kispert

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Contributions

K.K., M.L., and Y.Z. conducted experiments and contributed to interpretation of data and writing of the manuscript; D.C.W. performed and analysed echocardiographic studies with K.K and C.Y.; C.T.B. contributed to the interpretation of echocardiographic and cardiac physiology data; T.G. conducted all in situ hybridization experiments; A.K. provided unpublished Sfrp2 genomic DNA clones, contributed to the interpretation of in situ hybridization data and writing of the manuscript; G.G. conducted the BIAcore and pulldown assays shown in Fig. 2; C.Y. conducted some of the MI operations; T.-C.L. conducted qRT–PCR experiments; G.H. participated in constructing DNA expression vectors; D.S.G. designed and oversaw biochemical and cell biology experiments and contributed to the interpretation of data and writing of the manuscript; T.N.S. oversaw the project, designed experiments, interpreted data and wrote the manuscript.

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Correspondence to Daniel S. Greenspan or Thomas N. Sato.

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Kobayashi, K., Luo, M., Zhang, Y. et al. Secreted Frizzled-related protein 2 is a procollagen C proteinase enhancer with a role in fibrosis associated with myocardial infarction. Nat Cell Biol 11, 46–55 (2009). https://doi.org/10.1038/ncb1811

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