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Bidirectional tumor/stroma crosstalk promotes metastasis in mesenchymal colorectal cancer

Oncogene volume 39, pages 2453–2466 (2020)Cite this article

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Abstract

Patients with the mesenchymal subtype colorectal cancer (CRC) have a poor prognosis, in particular patients with stroma-rich tumors and aberrant SMAD4 expression. We hypothesized that interactions between SMAD4-deficient CRC cells and cancer-associated fibroblasts provide a biological explanation. In transwell invasion assays, fibroblasts increased the invasive capacity of SMAD4-deficient HT29 CRC cells, but not isogenic SMAD4-proficient HT29 cells. A TGF-β/BMP-specific array showed BMP2 upregulation by fibroblasts upon stimulation with conditioned medium from SMAD4-deficient CRC cells, while also stimulating their invasion. In a mouse model for experimental liver metastasis, the co-injection of fibroblasts increased metastasis formation of SMAD4-deficient CRC cells (p = 0.02) but not that of SMAD4-proficient CRC cells. Significantly less metastases were seen in mice co-injected with BMP2 knocked-down fibroblasts. Fibroblast BMP2 expression seemed to be regulated by TRAIL, a factor overexpressed in SMAD4-deficient CRC cells. In a cohort of 146 stage III CRC patients, we showed that patients with a combination of high stromal BMP2 expression and the loss of tumor SMAD4 expression had a significantly poorer overall survival (HR 2.88, p = 0.04). Our results suggest the existence of a reciprocal loop in which TRAIL from SMAD4-deficient CRC cells induces BMP2 in fibroblasts, which enhances CRC invasiveness and metastasis.

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Fig. 1: SMAD4-deficient HT29 cells show increased invasiveness in the presence of fibroblasts.
Fig. 2: Increased SMAD4-deficient HT29 invasion toward fibroblasts is BMP signaling dependent.
Fig. 3: Fibroblast-derived BMP2 increased liver metastasis formation in mice.
Fig. 4: TRAIL, a cytokine upregulated in SMAD4-deficient CRC cell lines, regulates BMP2 expression in fibroblasts.
Fig. 5: The combination of stromal high BMP2 expression and epithelial SMAD4 loss predicts poor prognosis.

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Author notes

  1. These authors contributed equally: Sarah Ouahoud, Philip W. Voorneveld, Lukas J. A. C. Hawinkels, James C. H. Hardwick

Authors and Affiliations

  1. Department of Gastroenterology & Hepatology, Leiden University Medical Center, Leiden, The Netherlands

    Sarah Ouahoud, Philip W. Voorneveld, Lennart R. A. van der Burg, Eveline S. M. de Jonge-Muller, Mark J. A. Schoonderwoerd, Madelon Paauwe, Thijs de Vos, Sophie de Wit, Lukas J.A.C. Hawinkels & James C. H. Hardwick

  2. Department of Surgery, Leiden University Medical Center, Leiden, The Netherlands

    Gabi W. van Pelt & Wilma E. Mesker

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Ouahoud, S., Voorneveld, P.W., van der Burg, L.R.A. et al. Bidirectional tumor/stroma crosstalk promotes metastasis in mesenchymal colorectal cancer. Oncogene 39, 2453–2466 (2020). https://doi.org/10.1038/s41388-020-1157-z

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