Rapid target gene validation in complex cancer mouse models using re-derived embryonic stem cells
Article first published online: 15 JAN 2014
© 2014 The Authors.
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
EMBO Molecular Medicine
Volume 6, Issue 2, pages 212–225, February 2014
How to Cite
Huijbers, I. J., Bin Ali, R., Pritchard, C., Cozijnsen, M., Kwon, M.-C., Proost, N., Song, J.-Y., de Vries, H., Badhai, J., Sutherland, K., Krimpenfort, P., Michalak, E. M., Jonkers, J. and Berns, A. (2014), Rapid target gene validation in complex cancer mouse models using re-derived embryonic stem cells. EMBO Mol Med, 6: 212–225. doi: 10.1002/emmm.201303297
- Issue published online: 7 FEB 2014
- Article first published online: 15 JAN 2014
- Manuscript Revised: 22 OCT 2013
- Manuscript Accepted: 22 OCT 2013
- Manuscript Received: 18 JUL 2013
- Dutch Cancer Society
- National Roadmap
- EuroSyStem and EurocanPlatform
- embryonic stem cells;
- mouse models;
Human cancers modeled in Genetically Engineered Mouse Models (GEMMs) can provide important mechanistic insights into the molecular basis of tumor development and enable testing of new intervention strategies. The inherent complexity of these models, with often multiple modified tumor suppressor genes and oncogenes, has hampered their use as preclinical models for validating cancer genes and drug targets. In our newly developed approach for the fast generation of tumor cohorts we have overcome this obstacle, as exemplified for three GEMMs; two lung cancer models and one mesothelioma model. Three elements are central for this system; (i) The efficient derivation of authentic Embryonic Stem Cells (ESCs) from established GEMMs, (ii) the routine introduction of transgenes of choice in these GEMM-ESCs by Flp recombinase-mediated integration and (iii) the direct use of the chimeric animals in tumor cohorts. By applying stringent quality controls, the GEMM-ESC approach proofs to be a reliable and effective method to speed up cancer gene assessment and target validation. As proof-of-principle, we demonstrate that MycL1 is a key driver gene in Small Cell Lung Cancer.
The GEMM-ESC approach describes and validates an improved method for generating mouse cancer models directly from embryonic stem cells. This technology speeds up the generation/modification of mouse models, while minimizing cost and breeding efforts.
- ESCs cultured in 2i medium show improved pluripotency and display equal genomic stability as ESCs cultured under classic conditions.
- Chimeric mice generated from GEMM-ESCs are equally susceptible to tumors as compared to the parental strain, provided the chimeras display coat color contribution upwards of 70%.
- Experimental cohorts are generated on-demand in less than 4 months without the need for any breeding.
- Controlled single copy integration of a transgene in the Col1a1 locus allows for robust reporter and oncogene expression in a mouse model for small cell lung cancer.
- Archived ESCs with complex genetic traits can serve as an important resource for the generation of new mouse models of cancer or for the validation of candidate cancer genes.