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.
Similar content being viewed by others
References
Rattner, A. et al. A family of secreted proteins contains homology to the cysteine-rich ligand-binding domain of frizzled receptors. Proc. Natl Acad. Sci. USA 94, 2859–2863 (1997).
Mayr, T. et al. Fritz: a secreted frizzled-related protein that inhibits Wnt activity. Mech. Dev. 63, 109–125 (1997).
Finch, P. W. et al. Purification and molecular cloning of a secreted, Frizzled-related antagonist of Wnt action. Proc. Natl Acad. Sci. USA 94, 6770–6775 (1997).
Melkonyan, H. S. et al. SARPs: a family of secreted apoptosis-related proteins. Proc. Natl Acad. Sci. USA 94, 13636–13641 (1997).
Salic, A. N., Kroll, K. L., Evans, L. M. & Kirschner, M. W. Sizzled: a secreted Xwnt8 antagonist expressed in the ventral marginal zone of Xenopus embryos. Development 124, 4739–4748 (1997).
Bodine, P. V. et al. The Wnt antagonist secreted frizzled-related protein-1 is a negative regulator of trabecular bone formation in adult mice. Mol. Endocrinol. 18, 1222–1237 (2004).
Barandon, L. et al. Involvement of FrzA/sFRP-1 and the Wnt/frizzled pathway in ischemic preconditioning. Circ. Res. 96, 1299–1306 (2005).
Satoh, W., Gotoh, T., Tsunematsu, Y., Aizawa, S. & Shimono, A. Sfrp1 and Sfrp2 regulate anteroposterior axis elongation and somite segmentation during mouse embryogenesis. Development 133, 989–999 (2006).
Suzuki, H. et al. Epigenetic inactivation of SFRP genes allows constitutive WNT signaling in colorectal cancer. Nature Genet. 36, 417–422 (2004).
Zou, H. et al. Aberrant methylation of secreted frizzled-related protein genes in esophageal adenocarcinoma and Barrett's esophagus. Int. J. Cancer 116, 584–91 (2005).
Lee, J. L., Lin, C. T., Chueh, L. L. & Chang, C. J. Autocrine/paracrine secreted Frizzled-related protein 2 induces cellular resistance to apoptosis: a possible mechanism of mammary tumorigenesis. J. Biol. Chem. 279, 14602–14609 (2004).
Zi, X. et al. Expression of Frzb/secreted Frizzled-related protein 3, a secreted Wnt antagonist, in human androgen-independent prostate cancer PC-3 cells suppresses tumor growth and cellular invasiveness. Cancer Res. 65, 9762–9770 (2005).
Dufourcq, P. et al. FrzA, a secreted frizzled related protein, induced angiogenic response. Circulation 106, 3097–3103 (2002).
Oshima, T. et al. Myeloma cells suppress bone formation by secreting a soluble Wnt inhibitor, sFRP-2. Blood 106, 3160–3165 (2005).
Kim, A. S., Anderson, S. A., Rubenstein, J. L., Lowenstein, D. H. & Pleasure, S. J. Pax-6 regulates expression of SFRP-2 and Wnt-7b in the developing CNS. J. Neurosci. 21, RC132 (2001).
Barandon, L. et al. Reduction of infarct size and prevention of cardiac rupture in transgenic mice overexpressing FrzA. Circulation 108, 2282–2289 (2003).
Roth, W. et al. Secreted Frizzled-related proteins inhibit motility and promote growth of human malignant glioma cells. Oncogene 19, 4210–4220 (2000).
Lee, C. S., Buttitta, L. A., May, N. R., Kispert, A. & Fan, C. M. SHH-N upregulates Sfrp2 to mediate its competitive interaction with WNT1 and WNT4 in the somitic mesoderm. Development 127, 109–118 (2000).
Kim, B. M., Buchner, G., Miletich, I., Sharpe, P. T. & Shivdasani, R. A. The stomach mesenchymal transcription factor Barx1 specifies gastric epithelial identity through inhibition of transient Wnt signaling. Dev. Cell 8, 611–622 (2005).
Polesskaya, A., Seale, P. & Rudnicki, M. A. Wnt signaling induces the myogenic specification of resident CD45+ adult stem cells during muscle regeneration. Cell 113, 841–852 (2003).
Lei, Q. et al. Wnt signaling inhibitors regulate the transcriptional response to morphogenetic Shh–Gli signaling in the neural tube. Dev. Cell 11, 325–337 (2006).
Mirotsou, M. et al. Secreted frizzled related protein 2 (Sfrp2) is the key Akt-mesenchymal stem cell-released paracrine factor mediating myocardial survival and repair. Proc. Natl Acad. Sci. USA 104, 1643–1648 (2007).
Lee, H. X., Ambrosio, A. L., Reversade, B. & De Robertis, E. M. Embryonic dorsal-ventral signaling: secreted frizzled-related proteins as inhibitors of tolloid proteinases. Cell 124, 147–159 (2006).
Muraoka, O. et al. Sizzled controls dorsoventral polarity by repressing cleavage of the Chordin protein. Nature Cell Biol. 8, 329–338 (2006).
De Robertis, E. M. & Kuroda, H. Dorsal-ventral patterning and neural induction in Xenopus embryos. Annu. Rev. Cell Dev. Biol. 20, 285–308 (2004).
Schier, A. F. & Talbot, W. S. Molecular genetics of axis formation in zebrafish. Annu. Rev. Genet. 39, 561–613 (2005).
Hopkins, D. R., Keles, S. & Greenspan, D. S. The bone morphogenetic protein 1/Tolloid-like metalloproteinases. Matrix Biol. 26, 508–523 (2007).
Ge, G. & Greenspan, D. S. BMP1 controls TGFβ1 activation via cleavage of latent TGFβ-binding protein. J. Cell Biol. 175, 111–120 (2006).
Kawano, Y. & Kypta, R. Secreted antagonists of the Wnt signalling pathway. J. Cell Sci. 116, 2627–2634 (2003).
Banyai, L. & Patthy, L. The NTR module: domains of netrins, secreted frizzled related proteins, and type I procollagen C-proteinase enhancer protein are homologous with tissue inhibitors of metalloproteases. Protein Sci. 8, 1636–1642 (1999).
Ge, G., Fernandez, C. A., Moses, M. A. & Greenspan, D. S. Bone morphogenetic protein 1 processes prolactin to a 17-kDa antiangiogenic factor. Proc. Natl Acad. Sci. USA 104, 10010–10015 (2007).
Cleutjens, J. P., Blankesteijn, W. M., Daemen, M. J. & Smits, J. F. The infarcted myocardium: simply dead tissue, or a lively target for therapeutic interventions. Cardiovasc. Res. 44, 232–241 (1999).
Swynghedauw, B. Molecular mechanisms of myocardial remodeling. Physiol. Rev. 79, 215–262 (1999).
Woessner, J. F., Jr. The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. Arch. Biochem. Biophys. 93, 440–447 (1961).
Berg, R. A. Determination of 3- and 4-hydroxyproline. Methods Enzymol. 82, 372–98 (1982).
Wodarz, A. & Nusse, R. Mechanisms of Wnt signaling in development. Annu. Rev. Cell Dev. Biol. 14, 59–88 (1998).
Kessler, E. & Adar, R. Type I procollagen C-proteinase from mouse fibroblasts. Purification and demonstration of a 55-kDa enhancer glycoprotein. Eur. J. Biochem. 186, 115–121 (1989).
Takahara, K. et al. Type I procollagen COOH-terminal proteinase enhancer protein: identification, primary structure, and chromosomal localization of the cognate human gene (PCOLCE). J. Biol. Chem. 269, 26280–26285 (1994).
Moali, C. et al. Substrate-specific modulation of a multisubstrate proteinase. C-terminal processing of fibrillar procollagens is the only BMP-1-dependent activity to be enhanced by PCPE-1. J. Biol. Chem. 280, 24188–24194 (2005).
Ge, G., Zhang, Y., Steiglitz, B. M. & Greenspan, D. S. Mammalian tolloid-like 1 binds procollagen C-proteinase enhancer protein 1 and differs from bone morphogenetic protein 1 in the functional roles of homologous protein domains. J. Biol. Chem. 281, 10786–19798 (2006).
Garrigue-Antar, L., Francois, V. & Kadler, K. E. Deletion of epidermal growth factor-like domains converts mammalian tolloid into a chordinase and effective procollagen C-proteinase. J. Biol. Chem. 279, 49835–49841 (2004).
Martyn, U. & Schulte-Merker, S. The ventralized ogon mutant phenotype is caused by a mutation in the zebrafish homologue of Sizzled, a secreted Frizzled-related protein. Dev. Biol. 260, 58–67 (2003).
Yabe, T. et al. Ogon/Secreted Frizzled functions as a negative feedback regulator of Bmp signaling. Development 130, 2705–2716 (2003).
Sun, Y. & Weber, K. T. Infarct scar: a dynamic tissue. Cardiovasc. Res. 46, 250–256 (2000).
Jugdutt, B. I. Ventricular remodeling after infarction and the extracellular collagen matrix: when is enough enough? Circulation 108, 1395–1403 (2003).
Jugdutt, B. I. Remodeling of the myocardium and potential targets in the collagen degradation and synthesis pathways. Curr. Drug Targets Cardiovasc. Haematol. Disord. 3, 1–30 (2003).
Scott, I. C. et al. Mammalian BMP-1/Tolloid-related metalloproteinases, including novel family member mammalian Tolloid-like 2, have differential enzymatic activities and distributions of expression relevant to patterning and skeletogenesis. Dev. Biol. 213, 283–300 (1999).
Pappano, W. N., Steiglitz, B. M., Scott, I. C., Keene, D. R. & Greenspan, D. S. Use of Bmp1/Tll1 doubly homozygous null mice and proteomics to identify and validate in vivo substrates of bone morphogenetic protein 1/tolloid-like metalloproteinases. Mol. Cell Biol. 23, 4428–4438 (2003).
Fisher, L. W., Stubbs, J. T., 3rd & Young, M. F. Antisera and cDNA probes to human and certain animal model bone matrix noncollagenous proteins. Acta Orthop. Scand. Suppl. 266, 61–65 (1995).
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).
Author information
Authors and Affiliations
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.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Information
Supplementary Information (PDF 886 kb)
Supplementary Information
Supplementary movie 1 (MOV 3823 kb)
Supplementary Information
Supplementary Movie 2 (MOV 4063 kb)
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ncb1811
- Springer Nature Limited
This article is cited by
-
Transcriptome analysis of cardiac endothelial cells after myocardial infarction reveals temporal changes and long-term deficits
Scientific Reports (2024)
-
Identification of PCPE-2 as the endogenous specific inhibitor of human BMP-1/tolloid-like proteinases
Nature Communications (2023)
-
Wnt Signaling in Heart Development and Regeneration
Current Cardiology Reports (2022)
-
PMCA4 inhibition does not affect cardiac remodelling following myocardial infarction, but may reduce susceptibility to arrhythmia
Scientific Reports (2021)
-
Exendin-4 Attenuates Remodeling in the Remote Myocardium of Rats After an Acute Myocardial Infarction by Activating β-Arrestin-2, Protein Phosphatase 2A, and Glycogen Synthase Kinase-3 and Inhibiting β-Catenin
Cardiovascular Drugs and Therapy (2021)