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Isolated limb perfusion based anti-p21ras gene therapy in a rat rhabdomyosarcoma
Boudewijn Van Etten1, Sandra T Van Tiel, Gisela Ambagtsheer
1Erasmus University Medical Centre-Daniel den Hoed Cancer Centre, Department of Surgical Oncology, PO Box 520,1 3008 AE Rotterdam, The Netherlands.
Background:
Inhibition of ras oncogene is a promising new strategy. Gene therapy against ras proved successful in human and murine tumour cell lines. Previously we demonstrated effective targeted transfection of tumour in a rat model by using an isolated limb perfusion (ILP) for the delivery of adenoviral vectors.
Materials And Methods:
This study explores the anti-tumour activity of an adenoviral construct encoding an intracellular single-chain antibody (scFv) against p21ras (Y28). In order to determine the influence of the ras status on the efficacy of the scFv, we used a wild-type rat rhabdomyosarcoma and its ras-oncogene transfectant, for in vitro studies. In vivo we used the ILP delivery method to study anti-tumour activity on established limb tumours.
Results:
In vitro studies demonstrated an inhibition of growth caused by the Y28 construct. No significant difference between transfected and wild-type cell lines could be demonstrated. Upon ILP, homogeneous transduction was observed in 5% of tumour cells. Perfusion with the Y28 construct, however, did not result in any additional anti-tumour activity compared to controls.
Conclusion:
Despite in vitro activity and in vivo transfection, no significant tumour response could be detected using anti-p21ras gene therapy in this ILP-tumour model.
Insights
Gene therapy targeting the ras oncogene showed in vitro growth inhibition but failed to demonstrate significant anti-tumour activity in vivo. Adenoviral vector delivery via isolated limb perfusion (ILP) did not improve outcomes for ras-targeted gene therapy.
Area of Science:
- Oncology
- Gene Therapy
- Molecular Biology
Background:
- Ras oncogene inhibition is a promising cancer therapy strategy.
- Gene therapy targeting ras has shown success in preclinical models.
- Isolated limb perfusion (ILP) is an effective method for targeted tumor transfection.
Purpose of the Study:
- To evaluate the anti-tumour activity of an adenoviral construct (Y28) encoding a single-chain antibody (scFv) against p21ras.
- To assess the influence of ras oncogene status on therapeutic efficacy.
- To investigate the in vivo anti-tumour effects using ILP delivery.
Main Methods:
- In vitro studies using wild-type and ras-oncogene-transfected rat rhabdomyosarcoma cell lines.
- In vivo studies utilizing ILP for adenoviral vector delivery to established limb tumors.
- Assessment of growth inhibition and tumour response.
Main Results:
- The Y28 construct inhibited cell growth in vitro, with no significant difference between wild-type and transfected cells.
- In vivo ILP achieved homogeneous transduction in 5% of tumor cells.
- No significant anti-tumour activity was observed with Y28 construct perfusion compared to controls.
Conclusions:
- Despite in vitro efficacy and successful in vivo transfection, anti-p21ras gene therapy via ILP did not yield significant tumour response.
- The study highlights limitations of this specific gene therapy approach in the tested model.
- Further research may be needed to optimize ras-targeted gene therapy strategies.
