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Dysregulated CRTC1 activity is a novel component of PGE2 signaling that contributes to colon cancer growth

Oncogene volume 35, pages 2602–2614 (2016)Cite this article

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Abstract

First identified as a dedicated CREB (cAMP response element-binding protein) co-activator, CRTC1 (CREB-regulated transcription co-activator 1) has been widely implicated in various neuronal functions because of its predominant expression in the brain. However, recent evidences converge to indicate that CRTC1 is aberrantly activated in an expanding number of adult malignancies. In this study, we provide strong evidences of enhanced CRTC1 protein content and transcriptional activity in mouse models of sporadic (APCmin/+ mice) or colitis-associated colon cancer azoxymethane/dextran sulfate sodium (AOM/DSS-treated mice), and in human colorectal tumors specimens compared with adjacent normal mucosa. Among signals that could trigger CRTC1 activation during colonic carcinogenesis, we demonstrate that treatment with cyclooxygenase 2 (COX2) inhibitors reduced nuclear CRTC1 active form levels in colonic tumors of APCmin/+ or AOM/DSS mice. In accordance, prostaglandins E2 (PGE2) exposure to human colon cancer cell lines promoted CRTC1 dephosphorylation and parallel nuclear translocation, resulting in enhanced CRTC1 transcriptional activity, through EP1 and EP2 receptors signaling and consecutive calcineurin and protein kinase A activation. In vitro CRTC1 loss of function in colon cancer cell lines was associated with reduced viability and cell division rate as well as enhanced chemotherapy-induced apoptosis on PGE2 treatment. Conversely, CRTC1 stable overexpression significantly increased colonic xenografts tumor growth, therefore demonstrating the role of CRTC1 signaling in colon cancer progression. Identification of the transcriptional program triggered by enhanced CRTC1 expression during colonic carcinogenesis, revealed some notable pro-tumorigenic CRTC1 target genes including NR4A2, COX2, amphiregulin (AREG) and IL-6. Finally, we demonstrate that COX2, AREG and IL-6 promoter activities triggered by CRTC1 are dependent on functional AP1 and CREB transcriptional partners. Overall, our study establishes CRTC1 as new mediator of PGE2 signaling, unravels the importance of its dysregulation in colon cancer and strengthens its use as a bona fide cancer marker.

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Acknowledgements

We gratefully acknowledge the Cancéropôle IdF funding source that made Yoann Schumacher PhD work possible. Otherwise, the studies were supported by grants from the Paris Descartes University, INSERM and the ARC research foundation against cancer. The authors are indebted to the Cochin Institute Transcriptomic Facility for technical help. The authors also thank (i) J Mathieu (Institut Cochin, Inserm U1016, Paris, France) for providing mouse BM-derived macrophage-conditioned media, (ii) Y Attie and D Vignjevic for providing human cancer associated fibroblasts conditioned media (Institut Curie, UMR144, Paris, France) and (ii) Ono Pharmaceutical Co. Ltd (Osaka, Japan) for providing PGE2 receptor subtype agonists. The authors are grateful to AF. Burnol (Institut Cochin, Inserm U1016, Paris, France), J Lebihan and M Legall (Centre de Recherche sur l'Inflammation Paris Montmartre, Inserm UMRS 1149, Paris, France), M Moldes (INSERM UMRS 938, Paris, France) and S Thenet (Centre de Recherche des Cordeliers, UMRS 1138, Paris, France) for critical reading of the manuscript.

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Authors and Affiliations

  1. Inserm U1016, Institut Cochin, Paris, France

    Y Schumacher, S Ourabah, F Baraille, R Dentin, C Postic & S Guilmeau

  2. CNRS UMR 8104, Paris, France

    Y Schumacher, S Ourabah, F Baraille, R Dentin, C Postic & S Guilmeau

  3. Université Paris Descartes, Paris, France

    Y Schumacher, S Ourabah, F Baraille, R Dentin, C Postic & S Guilmeau

  4. Université Paris Diderot, Paris, France

    Y Schumacher

  5. Gastroenterology and Digestive Oncology Unit, Avicenne Hospital, HUPSSD, APHP, Université Paris 13, Bobigny, France

    T Aparicio

  6. Pathology Unit, Avicenne Hospital, HUPSSD, APHP, Université Paris 13, Bobigny, France

    A Martin

  7. Digestive Surgery Unit, Avicenne Hospital, HUPSSD, APHP, Université Paris 13, Bobigny, France

    P Wind

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Schumacher, Y., Aparicio, T., Ourabah, S. et al. Dysregulated CRTC1 activity is a novel component of PGE2 signaling that contributes to colon cancer growth. Oncogene 35, 2602–2614 (2016). https://doi.org/10.1038/onc.2015.283

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