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Updated: May 1, 2026
Identification and Characterization of Metastatic Factors by Gene Transfer into the Novel RIP-Tag; RIP-tva Murine Model
Published on: October 16, 2017
Development of genetically flexible mouse models of sarcoma using RCAS-TVA mediated gene delivery
Leah Kabaroff1, Amar Gupta2, Serena Menezes1
1Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
Abstract:
Sarcomas are a heterogeneous group of mesenchymal malignancies and unfortunately there are limited functional genomics platforms to assess the molecular pathways contributing to sarcomagenesis. Thus, novel model systems are needed to validate which genes should be targeted for therapeutic intervention. We hypothesized that delivery of oncogenes into mouse skeletal muscle using a retroviral (RCAS-TVA) system would result in sarcomagenesis. We also sought to determine if the cell type transformed (mesenchymal progenitors vs. terminally differentiated tissues) would influence sarcoma biology. Cells transduced with RCAS vectors directing the expression of oncoproteins KrasG12D, c-Myc and/or Igf2 were injected into the hindlimbs of mice that expressed the retroviral TVA receptor in neural/mesenchymal progenitors, skeletal/cardiac muscle or ubiquitously (N-tva, AKE and BKE strains respectively). Disrupting the G1 checkpoint CDKN2 (p16/p19-/-) resulted in sarcoma in 30% of p16/p19-/- xN-tva mice with a median latency of 23 weeks (range 8-40 weeks). A similar incidence occurred in p16/p19-/- xBKE mice (32%), however, a shorter median latency (10.4 weeks) was observed. p16/p19-/- xAKE mice also developed sarcomas (24% incidence; median 9 weeks) yet 31% of mice also developed lung sarcomas. Gene-anchored PCR demonstrated retroviral DNA integration in 86% of N-tva, 93% of BKE and 88% of AKE tumors. KrasG12D was the most frequent oncogene isolated. Oncogene delivery by the RCAS-TVA system can generate sarcomas in mice with a defective cell cycle checkpoint. Sarcoma biology differed between the different RCAS models we created, likely due to the cell population being transformed. This genetically flexible system will be a valuable tool for sarcoma research.
Insights
Creating new mouse models for sarcoma research is crucial. The RCAS-TVA retroviral system effectively generates sarcomas by delivering oncogenes, aiding in the study of cancer development and potential therapies.
Area of Science:
- Oncology
- Genetics
- Molecular Biology
Background:
- Sarcomas are rare cancers with limited functional genomics platforms for studying their development.
- Novel model systems are needed to identify therapeutic targets for sarcomagenesis.
Purpose of the Study:
- To investigate the utility of the RCAS-TVA retroviral system for inducing sarcomagenesis in mice.
- To determine how the cell type targeted for transformation influences sarcoma biology.
Main Methods:
- Oncogenes (KrasG12D, c-Myc, Igf2) were delivered via RCAS vectors into mice with varying TVA receptor expression.
- The G1 cell cycle checkpoint was disrupted by deleting CDKN2 (p16/p19-/-).
- Tumor incidence, latency, and oncogene presence were analyzed in different mouse strains.
Main Results:
- Sarcomas developed in mice with a defective G1 checkpoint (p16/p19-/-) following oncogene delivery.
- Sarcoma incidence ranged from 24-32% across different mouse strains (N-tva, AKE, BKE).
- The cell type transformed influenced sarcoma biology, with the AKE model also yielding lung sarcomas.
Conclusions:
- The RCAS-TVA system, combined with cell cycle checkpoint disruption, can successfully generate sarcomas in mice.
- This system provides a flexible platform for studying sarcoma development and evaluating therapeutic strategies.
- Targeting specific cell populations influences sarcoma characteristics, offering insights into disease heterogeneity.
