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Updated: Oct 13, 2026
An Orthotopic Bladder Cancer Model for Gene Delivery Studies
Published on: December 1, 2013
In vivo adenovirus-mediated suicide gene therapy of orthotopic bladder cancer
M A Sutton1, C T Freund, S A Berkman
1Scott Department of Urology, Baylor College of Medicine, Houston, Texas 77030, USA.
Abstract:
These studies were undertaken to determine the feasibility, safety, and efficacy of suicide gene therapy using adenoviral-mediated herpes simplex virus thymidine kinase (ADV/RSV-tk) and the prodrug ganciclovir (GCV) in an orthotopic murine bladder cancer model. We utilized a replication-defective adenoviral construct containing the beta-galactosidase gene as a control and the herpes simplex virus thymidine kinase gene as the therapeutic vector under the transcription control of the Rous sarcoma virus long terminal repeat promoter. Intravesically created, orthotopic bladder tumors were established in syngeneic C3H/He female mice. India ink injection and beta-galactosidase studies were performed to determine if transurethral administration, direct tumor injection, or the combination was the most efficient route of virus administration. Optimal dosing of ADV/RSV-tk was determined by direct tumor injection with increasing viral doses and treatment with GCV. Treatment efficacy, long-term survival, and toxicity were determined in separate but similar controlled experiments. Growth curve studies demonstrated reliable tumor formation by 14 days. Direct transvesical tumor injection resulted in the best distribution and intratumor gene expression as measured by X-gal staining. Dose-ranging experiments demonstrated an optimal viral dose of 5 x 10(8) plaque-forming units and a greater than twofold reduction in tumor growth for the animals treated with ADV/RSV-tk compared to controls. Efficacy studies demonstrated a greater than threefold reduction in tumor growth. No clinical or gross pathologic toxicity was detected. Long-term survival results suggested a survival benefit for the treatment animals compared to controls. We conclude that ADV/RSV-tk in combination with GCV provides effective therapy for orthotopic murine bladder cancer by significantly inhibiting tumor growth with limited toxicity to the host. These data provide further support for testing this suicide gene therapy strategy in human Phase I trials.
Insights
This study shows that suicide gene therapy using adenoviral-mediated herpes simplex virus thymidine kinase (ADV/RSV-tk) and ganciclovir (GCV) effectively treats bladder cancer in mice. The therapy significantly reduced tumor growth with minimal toxicity, supporting human trials.
Area of Science:
- Oncology
- Gene Therapy
- Virology
Background:
- Bladder cancer remains a significant health concern, necessitating novel therapeutic strategies.
- Suicide gene therapy offers a targeted approach to cancer treatment.
- Adenoviral vectors have shown promise for gene delivery in cancer models.
Purpose of the Study:
- To evaluate the feasibility, safety, and efficacy of adenoviral-mediated suicide gene therapy.
- To assess the combination of herpes simplex virus thymidine kinase (ADV/RSV-tk) and ganciclovir (GCV) in an orthotopic murine bladder cancer model.
- To determine the optimal administration route and dosage for the therapeutic vector.
Main Methods:
- Orthotopic bladder tumors were established in syngeneic C3H/He mice.
- Replication-defective adenoviral vectors encoding HSV-tk or beta-galactosidase were used.
- Transurethral, direct tumor injection, or combination routes were compared for virus delivery.
- Dose-ranging studies determined optimal viral load, followed by efficacy and toxicity assessments.
Main Results:
- Direct transvesical tumor injection yielded the best viral distribution and gene expression.
- An optimal viral dose of 5 x 10(8) plaque-forming units was identified.
- ADV/RSV-tk plus GCV treatment resulted in a greater than threefold reduction in tumor growth.
- No significant clinical or pathological toxicity was observed, with a suggested survival benefit.
Conclusions:
- Adenoviral-mediated suicide gene therapy with ADV/RSV-tk and GCV is a feasible and effective treatment for orthotopic murine bladder cancer.
- The therapy demonstrates significant tumor growth inhibition with a favorable safety profile.
- These findings support the advancement of this suicide gene therapy strategy for human clinical trials.
