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Epithelium-derived Indian Hedgehog restricts stromal expression of ErbB family members that drive colonic tumor cell proliferation

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Abstract

Indian Hedgehog (Ihh) is a morphogen expressed by epithelial cells in the small intestine and colon that signals in a paracrine manner to gp38+ stromal cells. The loss of Ihh signaling results in increased epithelial proliferation, lengthening and multiplication of intestinal crypts and the activation of a stromal cell immune response. How Ihh controls epithelial proliferation through the stroma and how it affects colorectal cancer development remains poorly defined. To study the influence of Ihh signaling on the earliest stage of colorectal carcinogenesis, we used a well characterized mouse model in which both alleles of the Adenoma Polyposis Coli (Apc) gene could be inducibly deleted, leading to instant transformation of the colonic epithelium to an adenomatous phenotype. Concurrent deletion of Ihh from the adenomatous colonic epithelium of Apc inducible double mutant mice resulted in a remarkable increase in the hyperproliferative epithelial phenotype and increased accumulation of Lgr5+ stem cells. Transcriptional profiling of sorted colonic gp38+ fibroblasts showed upregulation of three ErbB pathway ligands (EREG, BTC, and NRG1) in Apc−/−Ihh−/− double mutant mice. We found that recombinant EREG, BTC, and NRG1 but not Lgr5 ligand R-Spondin promoted growth and proliferation of Apc double mutant colonic organoids. Thus, the loss of Ihh enhances Apc-driven colonic adenomagenesis via upregulation of ErbB pathway family members in colonic stromal cells. Our findings highlight the critical role of epithelium-derived Indian Hedgehog as a stromal tumor suppressor in the intestine.

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Fig. 1: Ihh acts to restrict hyperproliferation driven by the loss of Apc.
Fig. 2: Ihh acts to restrict expression of Lgr5+ stem cells upon loss of Apc and promotes differentiation.
Fig. 3: Ihh restricts activation of the WNT pathway downstream of the loss of Apc.
Fig. 4: Ihh restricts expression of ErbB family members in fibroblasts.
Fig. 5: Ihh suppresses expression of ErbB family members in fibroblasts.
Fig. 6: NRG1, EREG, and BTC enhance Apc mutant organoid growth.

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Acknowledgements

We thank our colleagues Berend Hooibrink and Toni van Capel from the Department of Experimental Immunology at the Amsterdam UMC, location AMC, for support with cell sorting, and the AMC Animal Research Facility (ARIA) for help with mouse experiments. We thank S.M. van Neerven and prof. dr. L. Vermeulen for providing Lgr5-EGFR-IRES-creERT2-Apcfl/fl colon organoids. This work was financially supported by a VICI grant (016140605) from the Netherlands Organization for Scientific Research (NWO) to GRB.

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Authors and Affiliations

  1. Tytgat Institute for Liver and Intestinal Research and Department of Gastroenterology & Hepatology, Amsterdam Gastroenterology and Metabolism, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands

    Florien Westendorp, Olga N. Karpus, Pim J. Koelink, Jacqueline L. M. Vermeulen, Sander Meisner, Nikè V. J. A. Büller, Manon E. Wildenberg, Vanesa Muncan & Gijs R. van den Brink

  2. Department of Oncogenomics, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands

    Jan Koster

  3. Roche Innovation Center Basel, F. Hoffmann-La Roche AG, Basel, Switzerland

    Gijs R. van den Brink

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  1. Florien Westendorp
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Contributions

Conceptualization and Methodology, BFW, ONK, VM, and GRB; Investigation, BFW, ONK, PJK, SM, and JLMV; Formal analysis, BFW, ONK, PJK, and VM; Resources, JK; Visualization, BFW, ONK, and VM; Writing—original draft, BFW and VM; Writing—review & editing, BFW, ONK, PJK, NVJAB, VM, MEW, GRB; Funding acquisition, GRB; Supervision, VM and GRB.

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Correspondence to Florien Westendorp.

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GRB is employee of Roche Innovation Center Basel, F. Hoffmann-La Roche AG, Basel, Switzerland.

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Westendorp, F., Karpus, O.N., Koelink, P.J. et al. Epithelium-derived Indian Hedgehog restricts stromal expression of ErbB family members that drive colonic tumor cell proliferation. Oncogene 40, 1628–1643 (2021). https://doi.org/10.1038/s41388-020-01633-0

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