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Autocrine WNT2 signaling in fibroblasts promotes colorectal cancer progression

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Abstract

The canonical WNT signaling pathway is crucial for intestinal stem cell renewal and aberrant WNT signaling is an early event in colorectal cancer (CRC) development. Here, we show for the first time that WNT2 is one of the most significantly induced genes in CRC stroma as compared to normal stroma. The impact of stromal WNT2 on carcinoma formation or progression was not addressed so far. Canonical WNT/β-catenin signaling was assessed using a 7TGP-reporter construct. Furthermore, effects of WNT2 on fibroblast migration and invasion were determined using siRNA-mediated gene silencing. Tumor cell invasion was studied using organotypic raft cultures and in vivo significance was assessed via a xenograft mouse model. We identified cancer-associated fibroblasts (CAFs) as the main source of WNT2. CAF-derived WNT2 activated canonical signaling in adenomatous polyposis coli/β-catenin wild-type colon cancer cells in a paracrine fashion, whereas no hyperactivation was detectable in cell lines harboring mutations in the adenomatous polyposis coli/β-catenin pathway. Furthermore, WNT2 activated autocrine canonical WNT signaling in primary fibroblasts, which was associated with a pro-migratory and pro-invasive phenotype. We identified FZD8 as the putative WNT2 receptor in CAFs. Three-dimensional organotypic co-culture assays revealed that WNT2-mediated fibroblast motility and extracellular matrix remodeling enhanced cancer cell invasion of cell lines even harboring mutations in the adenomatous polyposis coli/β-catenin pathway. Thus, suggesting a tumor-promoting influence on a broad range of CRC. In line, WNT2 also promotes tumor growth, invasion and metastasis in vivo. Moreover, high WNT2 expression is associated with poor prognosis in human CRC. The identification of the pro-malignant function of stromal derived WNT2 in CRC classifies WNT2 and its receptor as promising stromal targets to confine cancer progression in combination with conventional or targeted therapies.

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Acknowledgements

HD was supported by the Herzfelder Family Foundation and the Niederösterreichische Forschungs-und Bildungsges.m.b.H. AW was a recipient of a DOC Fellowship (24032) of the Austrian Academy of Sciences. RM was supported by the Austrian Science Fund (FWF, SFB F28 & F47). RE was supported by FWF (P25925-B20, P26908-B20, DK Inflammation and Immunity) and the Comprehensive Cancer Center (CCC) Vienna Research Grant.

Author contributions

NK and HD conceived and designed the research. NK, JS, CU, HN, AR, MS, DU, AW, MA, IC, TS, BP and JuSch performed the research. NK and HD analyzed the data and wrote the manuscript. MH, RE, XH, RM and WS were involved in data analysis and manuscript preparation. All authors reviewed and approved the manuscript for publication.

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Author notes

  1. H Nivarthi

    Present address: 10Current address: CeMM, Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, Vienna A-1090, Austria.,

  2. M Scherzer

    Present address: 11Current address: Karolinska Institutet, Solnavägen 1, Solna 171 77, Sweden.,

  3. N Kramer: This work contains parts of the PhD thesis of NK.

Authors and Affiliations

  1. Institute of Medical Genetics, Medical University of Vienna, Vienna, Austria

    N Kramer, J Schmöllerl, C Unger, D Unterleuthner, M Scherzer, A Riedl, T Schwarz, B Prieler, M Hengstschläger & H Dolznig

  2. Ludwig Boltzmann Institute for Cancer Research, Vienna, Austria

    J Schmöllerl, H Nivarthi & R Moriggl

  3. Boehringer Ingelheim RCV GmbH & Co KG, Vienna, Austria

    A Rudisch & W Sommergruber

  4. Max F. Perutz Laboratories, Medical University of Vienna, Vienna, Austria

    M Artaker

  5. Institute of Cancer Research, Medical University of Vienna, Vienna, Austria

    I Crncec & R Eferl

  6. Oncotest GmbH, Freiburg, Germany

    D Lenhardt & J Schüler

  7. Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA

    X Han

  8. Institute of Animal Breeding and Genetics, University of Veterinary Medicine Vienna, Vienna, Austria

    R Moriggl

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  1. N Kramer
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Correspondence to H Dolznig.

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Kramer, N., Schmöllerl, J., Unger, C. et al. Autocrine WNT2 signaling in fibroblasts promotes colorectal cancer progression. Oncogene 36, 5460–5472 (2017). https://doi.org/10.1038/onc.2017.144

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