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Directed evolution of adeno-associated virus for glioma cell transduction

  • Laboratory Investigation - Human/Animal Tissue
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Abstract

Glioblastoma multiforme (GBM) is a serious form of brain cancer for which there is currently no effective treatment. Alternative strategies such as adeno-associated virus (AAV) vector mediated-genetic modification of brain tumor cells with genes encoding anti-tumor proteins have shown promising results in preclinical models of GBM, although the transduction efficiency of these tumors is often low. As higher transduction efficiency of tumor cells should lead to enhanced therapeutic efficacy, a means to rapidly engineer AAV vectors with improved transduction efficiency for individual tumors is an attractive strategy. Here we tested the possibility of identifying high-efficiency AAV vectors for human U87 glioma cells by selection in culture of a newly constructed chimeric AAV capsid library generated by DNA shuffling of six different AAV cap genes (AAV1, AAV2, AAV5, AAVrh.8, AAV9, AAVrh.10). After seven rounds of selection, we obtained a chimeric AAV capsid that transduces U87 cells at high efficiency (97% at a dose of 104 genome copies/cell), and at low doses it was 1.45–1.6-fold better than AAV2, which proved to be the most efficient parental capsid. Interestingly, the new AAV capsid displayed robust gene delivery properties to all glioma cells tested (including primary glioma cells) with relative fluorescence indices ranging from 1- to 14-fold higher than AAV2. The selected vector should be useful for in vitro glioma research when efficient transduction of several cell lines is required, and provides proof-of-concept that an AAV library can be used to generate AAV vectors with enhanced transduction efficiency of glioma cells.

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Acknowledgments

This work was supported by a Young Investigator Award from the Alliance for Cancer Gene Therapy (MSE), NIH T32CA073479 (CM), and NIH P50CA86355 (MSE). We would like to acknowledge Dr. Rakesh Jain and the Edwin L. Steele Laboratory at MGH for support during this study. We would like to thank the MGH Nucleic Acid Quantitation Core facility supported by NINDS grant #P30NS4577 for the use of the thermal cyclers for quantitative PCR, and Drs. Johan Skog and Robert Carter for kindly providing the primary glioblastoma cells. We thank Johanna Niers for help with cell culture. We would like to thank Dr. Xandra O. Breakefield for advice and critical reading of the manuscript.

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Author notes

  1. Miguel Sena-Esteves

    Present address: Department of Neurology and Gene Therapy Center, University of Massachusetts Medical School, 381 Plantation Street, Suite 250, Worcester, MA, 01605, USA

Authors and Affiliations

  1. Department of Neurology, Massachusetts General Hospital, and Neuroscience Program, Harvard Medical School, Boston, MA, USA

    Casey A. Maguire, Davide Gianni, Lev A. Shaket & Miguel Sena-Esteves

  2. Imperial College, National Heart and Lung Institute, London, UK

    Davide Gianni

  3. Department of Neuroscience and Pharmacology, Rudolf Magnus Institute of Neuroscience, UMC Utrecht, Utrecht, The Netherlands

    Dimphna H. Meijer

  4. Department of Neurosurgery, Massachusetts General Hospital, Boston, MA, USA

    Hiroaki Wakimoto & Samuel D. Rabkin

  5. University of Massachusetts Medical School, and Gene Therapy Center, Worcester, MA, USA

    Guangping Gao

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  1. Casey A. Maguire
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  2. Davide Gianni
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Correspondence to Miguel Sena-Esteves.

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Maguire, C.A., Gianni, D., Meijer, D.H. et al. Directed evolution of adeno-associated virus for glioma cell transduction. J Neurooncol 96, 337–347 (2010). https://doi.org/10.1007/s11060-009-9972-7

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  • DOI: https://doi.org/10.1007/s11060-009-9972-7

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