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Combining high selectivity of replication with fiber chimerism for effective adenoviral oncolysis of CAR-negative melanoma cells

Gene Therapy volume 11, pages 1694–1702 (2004)Cite this article

Abstract

Oncolytic adenoviruses constitute a new and promising tool for cancer treatment that has been rapidly translated into clinical trials. However, minimal or absent expression of the adenovirus serotype 5 (Ad5) receptor CAR (coxsackievirus and adenovirus receptor) on cancer cells represents a major limitation for Ad5-based oncolysis. Here, we report on the resistance of CAR-negative primary melanoma cells to cell killing by wild-type Ad5 (Ad5wt) even after high titer infection, thus underlining the need for tropism-modification of oncolytic adenoviruses. We engineered a new generation of oncolytic adenoviruses that exhibit both efficient target cell infection by swapping Ad5 fiber domains with those of Ad serotype 3, which binds to a receptor distinct from CAR, and targeted virus replication. Fiber chimerism resulted in efficient cytopathicity to primary melanoma cells, which was at least 104-fold increased relative to Ad5wt. Since viral infectivity mediated by such modified viral capsids was not cell type-specific, it was pivotal to carefully restrict adenoviral replication to target cells. Towards this end, we replaced both E1A and E4 promoters of fiber chimeric viruses by tyrosinase enhancer/promoter constructs. The resulting viruses showed melanoma-specific expression of E1A and E4 and combined efficient virus replication and cell killing in melanoma cell lines and primary melanoma cells with a remarkable specificity profile that implements strong attenuation in nonmelanoma cells, including normal fibroblasts and keratinocytes.

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Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft (grant NE832/1 to DMN, and Graduiertenkolleg 592 to SS), the Deutsche Krebshilfe (grant 10-2186-Ne 1 to DMN) and the National Cancer Institute (grants R01 CA83821, P50 CA83591, R01 CA93796, and R01 CA94084). We are grateful to Dr D Dieckmann, Dr IJ Fidler, Dr F Noya, Dr J Price, L Rivera, Dr J Schlom, and Dr T Strong for cell lines and primary cells, to Dr J Chrobozcek for providing plasmid pBR.Ad3Fib, Dr Ruben Hernandez-Alcoceba for plasmid pUC19-E4P-, Dr Gary Ketner for the E4 antibody, and to T Uil for plasmid pNEB.PK.SnaBI.

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Author notes

  1. V Krasnykh

    Present address: Department of Experimental Diagnostic Imaging, MD Anderson Cancer Center, University of Texas, Houston, TX, USA

Authors and Affiliations

  1. Division of Human Gene Therapy, Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA

    AA Rivera, J Davydova, M Wang, V Krasnykh, M Yamamoto, DT Curiel & DM Nettelbeck

  2. Division of Human Gene Therapy, Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA

    AA Rivera, J Davydova, M Wang, V Krasnykh, M Yamamoto, DT Curiel & DM Nettelbeck

  3. Division of Human Gene Therapy, Department of Surgery, University of Alabama at Birmingham, Birmingham, AL, USA

    AA Rivera, J Davydova, M Wang, V Krasnykh, M Yamamoto, DT Curiel & DM Nettelbeck

  4. The Gene Therapy Center, University of Alabama at Birmingham, Birmingham, AL, USA

    AA Rivera, J Davydova, M Wang, V Krasnykh, M Yamamoto, DT Curiel & DM Nettelbeck

  5. Department of Dermatology, University Medical Center Erlangen, Erlangen, Germany

    S Schierer & DM Nettelbeck

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Rivera, A., Davydova, J., Schierer, S. et al. Combining high selectivity of replication with fiber chimerism for effective adenoviral oncolysis of CAR-negative melanoma cells. Gene Ther 11, 1694–1702 (2004). https://doi.org/10.1038/sj.gt.3302346

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  • DOI: https://doi.org/10.1038/sj.gt.3302346

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