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A next-generation dual-recombinase system for time- and host-specific targeting of pancreatic cancer

An Erratum to this article was published on 07 April 2015

This article has been updated

Abstract

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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Figure 1: Pdx1-Flp–activated expression of oncogenic KrasG12D induces premalignant PanIN and PDAC.
Figure 2: Secondary genetic manipulation of established KrasG12D-induced PanIN lesions and PDAC cells in the Pdx1-Flp lineage.
Figure 3: Validation of therapeutic targets in vivo by Cre-induced time-specific Pdpk1 inactivation or DTA-mediated tumor cell depletion.
Figure 4: Mast cells are dispensable for PDAC development.
Figure 5: Applications of DRS.

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  • 31 October 2014

     In the version of this article initially published online, affiliation 12 was missing for Roland Rad, Roland M. Schmid and Dieter Saur. The list should have read: “Roland Rad1,5,12,13, Roland M Schmid1,12,13 & Dieter Saur1,12,13.” The error has been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

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.).

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B.S. and D.S. designed research; N.S., B.S., K.S., C.V., C.S., M.Z., S.E., M.C.P., P.E., S.K., R.B., F.Y., A.S., I.V., R.R., G.S. and D.S., performed research; T.B.F., A.M.L., C.-L.L., E.J.M., D.G.K., A.S., D.R.A., I.V., A.B., A.K., A.E.S., H.-R.R., R.R. and R.M.S. contributed new reagents/analytic tools; N.S., B.S., K.S., C.V., C.S., M.Z., S.E., M.C.P., P.E., S.K., R.B., F.Y., I.V., R.R., G.S. and D.S. analyzed data; and B.S. and D.S. wrote the paper. N.S., B.S., K.S. and C.V. contributed equally to this manuscript.

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Correspondence to Dieter Saur.

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Supplementary Figures 1–11, Supplementary Table 1 and Supplementary Results (PDF 8865 kb)

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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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