Short Communication
Gene Therapy (2009) 16, 1042–1049; doi:10.1038/gt.2009.54; published online 4 June 2009
Efficient nonviral Sleeping Beauty transposon-based TCR gene transfer to peripheral blood lymphocytes confers antigen-specific antitumor reactivity
P D Peng1, C J Cohen1, S Yang1, C Hsu1, S Jones1, Y Zhao1, Z Zheng1, S A Rosenberg1 and R A Morgan1
1Surgery Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA
Correspondence: Dr RA Morgan, National Cancer Institute, 10 Center Drive, Building 10, CRC 3W-3864, Bethesda, MD, 20892, USA. E-mail: rmorgan@mail.nih.gov
Received 31 July 2008; Revised 12 November 2008; Accepted 10 December 2008; Published online 4 June 2009.
Abstract
Genetically engineered lymphocytes hold promise for the treatment of genetic disease, viral infections and cancer. However, current methods for genetic transduction of peripheral blood lymphocytes rely on viral vectors, which are hindered by production and safety-related problems. In this study, we demonstrated an efficient novel nonviral platform for gene transfer to lymphocytes. The Sleeping Beauty transposon-mediated approach allowed for long-term stable expression of transgenes at 
50% efficiency. Utilizing transposon constructs expressing tumor antigen-specific T-cell receptor genes targeting p53 and MART-1, we demonstrated sustained expression and functional reactivity of transposon-engineered lymphocytes on encountering target antigen presented on tumor cells. We found that transposon- and retroviral-modified lymphocytes had comparable transgene expression and phenotypic function. These results demonstrate the promise of nonviral ex vivo genetic modification of autologous lymphocytes for the treatment of cancer and immunologic disease.
Keywords:
Sleeping Beauty transposon, TCR gene therapy, adoptive immunotherapy, nonviral vector, T-cell gene transfer
