Genetically engineered mouse models (GEMMs) have dramatically improved our understanding of tumor evolution and therapeutic resistance. However, sequential genetic manipulation of gene expression and targeting of the host is almost impossible using conventional Cre-loxP–based models. We have developed an inducible dual-recombinase system by combining flippase-FRT (Flp-FRT) and Cre-loxP recombination technologies to improve GEMMs of pancreatic cancer. This enables investigation of multistep carcinogenesis, genetic manipulation of tumor subpopulations (such as cancer stem cells), selective targeting of the tumor microenvironment and genetic validation of therapeutic targets in autochthonous tumors on a genome-wide scale. As a proof of concept, we performed tumor cell–autonomous and nonautonomous targeting, recapitulated hallmarks of human multistep carcinogenesis, validated genetic therapy by 3-phosphoinositide-dependent protein kinase inactivation as well as cancer cell depletion and show that mast cells in the tumor microenvironment, which had been thought to be key oncogenic players, are dispensable for tumor formation.
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The authors thank A. Berns (Netherlands Cancer Institute), S. Dymecki (Harvard Medical School), T. Jacks (Massachusetts Institute of Technology), L. Luo (Stanford University), J. Martinez-Barbera (University College London) and D. Tuveson (Cold Spring Harbor Laboratory) for providing transgenic animals, C. Wright (Vanderbilt University) for the mouse Pdx1 promoter construct, P. Soriano (Mount Sinai School of Medicine) for the Flp-o expression vector and the R26 targeting vector, T. Schmidt and M. Bewerunge-Hudler (DKFZ Microarray Core Facility) for mRNA analyses, and J. Götzfried, U. Götz and S. Jaeckel for technical assistance. This work was supported by funding from Deutsche Forschungsgemeinschaft (DFG SA 1374/4-1 to D.S. and SFB824, TP C9 to G.S. and D.S.), the Helmholtz Alliance Preclinical Comprehensive Cancer Center (to H.-R.R., R.R., R.M.S. and D.S.), the German Cancer Consortium (DKTK) (to R.R., R.M.S. and D.S.), the Wilhelm-Sander Foundation (2012.084.1 to G.S.), the Spanish Ministerio de Economía y Competitividad subprograma Ramón y Cajal (I.V.), the European Union (ERC Advanced Grant No.233074 to H.-R.R.), and the National Cancer Institute USA (R01 CA138265 to D.G.K. and CA155620 to A.M.L.).
The authors declare no competing financial interests.
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Schönhuber, N., Seidler, B., Schuck, K. et al. A next-generation dual-recombinase system for time- and host-specific targeting of pancreatic cancer. Nat Med 20, 1340–1347 (2014). https://doi.org/10.1038/nm.3646
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