Abstract
In multicellular organisms, a tight control of cell death is required to ensure normal development and tissue homeostasis. Improper function of apoptotic or survival pathways can not only affect developmental programs but also favor cancer progression. Here we describe a novel apoptotic signaling pathway involving the transmembrane receptor Kremen1 and its ligand, the Wnt-antagonist Dickkopf1. Using a whole embryo culture system, we first show that Dickkopf1 treatment promotes cell survival in a mouse model exhibiting increased apoptosis in the developing neural plate. Remarkably, this effect was not recapitulated by chemical Wnt inhibition. We then show that Dickkopf1 receptor Kremen1 is a bona fide dependence receptor, triggering cell death unless bound to its ligand. We performed Wnt-activity assays to demonstrate that the pro-apoptotic and anti-Wnt functions mediated by Kremen1 are strictly independent. Furthermore, we combined phylogenetic and mutagenesis approaches to identify a specific motif in the cytoplasmic tail of Kremen1, which is (i) specifically conserved in the lineage of placental mammals and (ii) strictly required for apoptosis induction. Finally, we show that somatic mutations of kremen1 found in human cancers can affect its pro-apoptotic activity, supporting a tumor suppressor function. Our findings thus reveal a new Wnt-independent function for Kremen1 and Dickkopf1 in the regulation of cell survival with potential implications in cancer therapies.
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Abbreviations
- CNS:
-
central nervous system
- COSMIC:
-
catalogue of somatic mutations in cancer
- DCC:
-
deleted in colorectal cancer
- Dkk:
-
Dickkopf
- ECD:
-
extracellular domain
- GFP:
-
green fluorescent protein
- HA:
-
hemagglutinin
- HEK:
-
human embryonic kidney
- ICD:
-
intracellular domain
- Krm:
-
Kringle-coding gene marking the eye and the nose
- LRP:
-
low-density lipoprotein receptor-related protein
- TCGA:
-
the cancer genome atlas
- TUNEL:
-
terminal deoxynucleotidyl transferase dUTP nick end labeling
- Wnt:
-
wingless-related integration site
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Acknowledgements
We are grateful to Thierry Galli, Anne Camus, Stéphane Nedelec, Eva Coppola and Sonia Garel for helpful discussions and critical reading of the manuscript. We wish to thank Anne Camus and Yoko Arai for their help in implementing whole embryo cultures, Christine Vesque and Sylvie Schneider-Maunoury for providing zebrafish cDNA, De-Li Shi for providing xenopus cDNA, Thierry Galli for providing HEK cells, Eve Gazave for her help regarding pharmacological agents, Anne-Laure Todeschini for her help with luciferase assays, Fabien Fauchereau for TCGA data mining, Lisa Vigier and Annie Dutriaux for technical assistance, Betty Freret-Hodara for helpful comments, Elenat Marot, Natalia Maties and Sébastien Gravat for maintaining mouse colonies, as well as the ImagoSeine imaging facility (member of the France BioImaging infrastructure, supported by the French National Research Agency ANR-10-INSB-04 ‘Investments for the future’). FC is an Inserm researcher. AP is a CNRS investigator and member Team of the Ecole des Neurosciences de Paris (ENP). This work was supported by grants from the Agence Nationale de la Recherche (ANR-2011-BSV4-023-01), the Fondation pour la Recherche Médicale (FRM) (INE20060306503 and Equipe FRM DEQ20130326521), Ville de Paris (2006 ASES 102) and the Association pour la Recherche sur le Cancer (ARC) (SFI 2011 1203674) to AP and Comité de Paris de la Ligue contre le cancer (RS14/75-7 and RS15/75-46) to FC.
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Causeret, F., Sumia, I. & Pierani, A. Kremen1 and Dickkopf1 control cell survival in a Wnt-independent manner. Cell Death Differ 23, 323–332 (2016). https://doi.org/10.1038/cdd.2015.100
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DOI: https://doi.org/10.1038/cdd.2015.100
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