Sequential gene targeting to make chimeric tumor models with de novo chromosomal abnormalities
Jennifer S Chambers1, Tomoyuki Tanaka, Tim Brend
1Authors' Affiliations: MRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford, Oxford; Leeds Institute of Molecular Medicine, Wellcome Trust Brenner Building, St. James's University Hospital, University of Leeds, Leeds, United Kingdom; and Molecular Cytogenetics Group, Spanish National Cancer Research Center (CNIO), Melchor Fernandez Almagro, Madrid, Spain.
This study introduces a novel gene targeting method in embryonic stem cells to create cancer models in mice. This approach accelerates research by reducing mouse numbers and enabling the study of genetic interactions in cancer development.
Area of Science:
- Genetics
- Cancer Biology
- Genomic Medicine
Background:
- Chromosomal abnormalities are hallmarks of various cancers, including leukemia, lymphoma, sarcomas, and epithelial tumors.
- Next-generation sequencing has identified numerous genetic alterations, including chromosomal abnormalities and mutations, in cancer genomes.
- Current mouse models often require multiple genetic modifications, posing limitations for studying complex genetic interactions in cancer development.
Purpose of the Study:
- To develop an accelerated and efficient technology for generating genetically defined cancer models in mice.
- To overcome the limitations of traditional mouse models in assessing the interrelationship of genetic abnormalities and mutations in cancer.
- To provide a flexible platform for investigating the role of chromosomal abnormalities and mutations in cell-specific cancer initiation and progression.
Main Methods:
- Sequential gene targeting of embryonic stem cells to introduce multiple genetic alterations.
- Generation of chimeric offspring from genetically modified progenitor cells.
- Germline transmission of sequentially targeted alleles to confirm genome integrity.
- Fluorescence tagging of cancer-initiating cells for visualization and tracking.
Main Results:
- Demonstrated that MLL-ENL fusion is sufficient to induce lethal leukocytosis in chimeric mice.
- Validated the technology through germline transmission, confirming the integrity of the targeted genome.
- Showcased the ability to generate progenitor cells carrying all necessary genetic changes for cell-specific cancer models.
- Highlighted the reduction in mouse numbers required, aligning with the 3Rs principles (Replacement, Reduction, Refinement).
Conclusions:
- Sequential gene targeting in embryonic stem cells offers an accelerated platform for creating complex cancer models.
- This technology facilitates the study of gene interactions and the role of chromosomal abnormalities in cancer.
- The method reduces animal usage and provides valuable tools for cancer research, including the identification of cancer-initiating cells.
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