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Published on: October 6, 2022
Targeting of adenovirus to human renal cell carcinoma cells
Wim Jongmans1, Kelly van den Oudenalder, Dorien M Tiemessen
1Department of Experimental Urology, Nijmegen Center for Molecular Life Sciences, University Medical Center Nijmegen, Nijmegen, The Netherlands.
Objectives:
The use of recombinant adenoviruses in cancer gene therapy is limited by the widespread expression of the coxsackievirus and adenovirus receptor on normal human cells. Targeting adenoviral vectors to renal cell carcinoma (RCC) cells may improve their potential in cancer gene therapy of patients with metastatic RCC. The G250 protein, also known as the carbonic anhydrase IX protein, is membranously expressed in all cases of clear cell RCC, and clinical studies have demonstrated exceptional tumor targeting with a G250 monoclonal antibody.
Methods:
A recombinant bispecific single-chain antibody directed against the RCC-associated G250 protein and the adenovirus fiber knob domain was constructed and used to retarget recombinant adenovirus expressing the green fluorescent protein under control of the cytomegalovirus promoter. G250-specific adenoviral transduction of cells was examined by flow cytometric analysis of green fluorescent protein expression.
Results:
G250-positive RCC cells displayed enhanced susceptibility for transduction by the green fluorescent protein recombinant adenovirus complexed with the G250-directed bispecific single-chain antibody when compared with native adenovirus. This enhanced transduction was restricted to G250-positive RCC cells and could be abolished completely in the presence of excess G250 protein.
Conclusions:
The results of this study demonstrate the feasibility of immunologic retargeting of adenovirus to RCC cells with the highly tumor-specific G250 protein as the target. This strategy may provide the possibility of improving cancer gene therapy for patients with RCC.
Insights
This study retargeted adenoviruses using a bispecific antibody to target G250-positive renal cell carcinoma (RCC) cells. This approach enhances gene therapy delivery specifically to cancer cells, improving treatment potential for metastatic RCC patients.
Area of Science:
- Oncolytic virotherapy
- Gene therapy vectors
- Immunology
Background:
- Adenoviral vectors for cancer gene therapy are limited by coxsackievirus and adenovirus receptor expression on normal cells.
- Targeting adenoviral vectors to renal cell carcinoma (RCC) can improve gene therapy efficacy.
- G250 (carbonic anhydrase IX) is highly expressed on clear cell RCC, showing promise for tumor targeting.
Purpose of the Study:
- To retarget adenoviral vectors to RCC cells using the G250 tumor-specific antigen.
- To evaluate the efficacy of a bispecific antibody in directing adenoviral transduction to G250-positive RCC cells.
Main Methods:
- Constructed a bispecific single-chain antibody targeting G250 and the adenovirus fiber knob.
- Used the antibody to retarget adenovirus expressing green fluorescent protein (GFP).
- Assessed G250-specific adenoviral transduction via GFP expression using flow cytometry.
Main Results:
- G250-positive RCC cells showed significantly enhanced transduction with retargeted adenovirus compared to native adenovirus.
- Enhanced transduction was specific to G250-positive cells.
- Transduction was completely blocked by excess G250 protein, confirming specificity.
Conclusions:
- Demonstrated the feasibility of immunologic retargeting of adenovirus to RCC using the G250 protein.
- This strategy holds potential for improving cancer gene therapy outcomes in RCC patients.
