Rapid target gene validation in complex cancer mouse models using re‐derived embryonic stem cells

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 gene...

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Bibliographic Details
Published in:EMBO molecular medicine 2014-02, Vol.6 (2), p.212-225
Main Authors: Huijbers, Ivo J, Bin Ali, Rahmen, Pritchard, Colin, Cozijnsen, Miranda, Kwon, Min‐Chul, Proost, Natalie, Song, Ji‐Ying, Vries, Hilda, Badhai, Jitendra, Sutherland, Kate, Krimpenfort, Paul, Michalak, Ewa M, Jonkers, Jos, Berns, Anton
Format: Article
Language:English
Subjects:
Animal models
Animals
cancer
Carcinogenesis - metabolism
Carcinogenesis - pathology
Cell Proliferation
Cells, Cultured
Chimera
chimeras
Clone Cells
Cloning
Disease Models, Animal
DNA Nucleotidyltransferases - metabolism
Embryo cells
embryonic stem cells
Embryonic Stem Cells - cytology
Embryonic Stem Cells - metabolism
Experiments
FLP recombinase
Gene Transfer Techniques
Genes
Genes, Reporter
Genetic engineering
Genomic Instability
Genomics
Genotype
Germ Cells - metabolism
Humans
Luciferases - metabolism
Lung cancer
Lung Neoplasms - metabolism
Lung Neoplasms - pathology
Medical research
Mesothelioma
Mice
Mice, Inbred C57BL
mouse models
Mutation
MycL1
Oncogenes
Phenotype
Pluripotent Stem Cells - metabolism
Quality Control
Reproducibility of Results
Small cell lung carcinoma
Small Cell Lung Carcinoma - metabolism
Small Cell Lung Carcinoma - pathology
Stem cell transplantation
Stem cells
Transgenes
Tumor suppressor genes
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Summary: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. Synopsis 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. 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.
ISSN:1757-4676
1757-4684
DOI:10.1002/emmm.201303297