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Nat Med. 2012 May;18(5):807-815. doi: 10.1038/nm.2700.

Editing T cell specificity towards leukemia by zinc finger nucleases and lentiviral gene transfer.

Author information

1
Experimental Hematology Unit, Division of Regenerative Medicine, Gene Therapy and Stem Cells, Program of Immunology, Gene Therapy and Bio-Immunotherapy of Cancer, Leukemia Unit, San Raffaele Scientific Institute, Milan, Italy.
2
Vita Salute San Raffaele University, Milan, Italy.
3
San Raffaele Telethon Institute for Gene Therapy, Division of Regenerative Medicine, Gene Therapy and Stem Cells, San Raffaele Scientific Institute, Milan, Italy.
4
Sangamo BioSciences Inc., Richmond, USA.
5
Fred Hutchinson Cancer Research Center and University of Washington, Seattle, USA.
6
Experimental Immunology Unit, Division of Immunology, Transplantation and Infectious Diseases, Program of Immunology, Gene Therapy and Bio-Immunotherapy of Cancer, San Raffaele Scientific Institute, Milan, Italy.
7
Department of Pathology, Leukemia Unit, San Raffaele Scientific Institute, Milan, Italy.
8
Hematology Clinical Unit, Division of Regenerative Medicine, Gene Therapy and Stem Cells, San Raffaele Scientific Institute, Milan, Italy.
#
Contributed equally

Abstract

The transfer of high-avidity T cell receptor (TCR) genes isolated from rare tumor-specific lymphocytes into polyclonal T cells is an attractive cancer immunotherapy strategy. However, TCR gene transfer results in competition for surface expression and inappropriate pairing between the exogenous and endogenous TCR chains, resulting in suboptimal activity and potentially harmful unpredicted antigen specificities of the resultant TCRs. We designed zinc-finger nucleases (ZFNs) that promoted the disruption of endogenous TCR β- and α-chain genes. Lymphocytes treated with ZFNs lacked surface expression of CD3-TCR and expanded with the addition of interleukin-7 (IL-7) and IL-15. After lentiviral transfer of a TCR specific for the Wilms tumor 1 (WT1) antigen, these TCR-edited cells expressed the new TCR at high levels, were easily expanded to near purity and were superior at specific antigen recognition compared to donor-matched, unedited TCR-transferred cells. In contrast to unedited TCR-transferred cells, the TCR-edited lymphocytes did not mediate off-target reactivity while maintaining their anti-tumor activity in vivo, thus showing that complete editing of T cell specificity generates tumor-specific lymphocytes with improved biosafety profiles.

PMID:
22466705
PMCID:
PMC5019824
DOI:
10.1038/nm.2700
[Indexed for MEDLINE]
Free PMC Article

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