Journal of Neuro-Oncology

, Volume 96, Issue 3, pp 337–347

Directed evolution of adeno-associated virus for glioma cell transduction

  • Casey A. Maguire
  • Davide Gianni
  • Dimphna H. Meijer
  • Lev A. Shaket
  • Hiroaki Wakimoto
  • Samuel D. Rabkin
  • Guangping Gao
  • Miguel Sena-Esteves
Laboratory Investigation - Human/Animal Tissue

DOI: 10.1007/s11060-009-9972-7

Cite this article as:
Maguire, C.A., Gianni, D., Meijer, D.H. et al. J Neurooncol (2010) 96: 337. doi:10.1007/s11060-009-9972-7

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.

Keywords

Adeno-associated virus (AAV)LibraryBrain tumorGlioblastomaGene therapyGene transfer

Supplementary material

11060_2009_9972_MOESM2_ESM.pdf (210 kb)
(PDF 210 kb)

Copyright information

© Springer Science+Business Media, LLC. 2009

Authors and Affiliations

  • Casey A. Maguire
    • 1
  • Davide Gianni
    • 1
    • 2
  • Dimphna H. Meijer
    • 3
  • Lev A. Shaket
    • 1
  • Hiroaki Wakimoto
    • 4
  • Samuel D. Rabkin
    • 4
  • Guangping Gao
    • 5
  • Miguel Sena-Esteves
    • 1
    • 6
  1. 1.Department of Neurology, Massachusetts General Hospital, and Neuroscience ProgramHarvard Medical SchoolBostonUSA
  2. 2.Imperial College, National Heart and Lung InstituteLondonUK
  3. 3.Department of Neuroscience and PharmacologyRudolf Magnus Institute of Neuroscience, UMC UtrechtUtrechtThe Netherlands
  4. 4.Department of NeurosurgeryMassachusetts General HospitalBostonUSA
  5. 5.University of Massachusetts Medical School, and Gene Therapy CenterWorcesterUSA
  6. 6.Department of Neurology and Gene Therapy CenterUniversity of Massachusetts Medical SchoolWorcesterUSA