title: Editing T cell specificity towards leukemia by zinc finger nucleases and lentiviral gene transfer.
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.
authors.affiliation: 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.|Vita Salute San Raffaele University, Milan, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Provasi
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy, Division of Regenerative Medicine, Gene Therapy and Stem Cells, San Raffaele Scientific Institute, Milan, Italy.|Vita Salute San Raffaele University, Milan, Italy.
authors.forename: Pietro
authors.initials: P
authors.lastname: Genovese
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy, Division of Regenerative Medicine, Gene Therapy and Stem Cells, San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Angelo
authors.initials: A
authors.lastname: Lombardo
authors.affiliation: 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.
authors.forename: Zulma
authors.initials: Z
authors.lastname: Magnani
authors.affiliation: Sangamo BioSciences Inc., Richmond, USA.
authors.forename: Pei-Qi
authors.initials: PQ
authors.lastname: Liu
authors.affiliation: Sangamo BioSciences Inc., Richmond, USA.
authors.forename: Andreas
authors.initials: A
authors.lastname: Reik
authors.affiliation: Sangamo BioSciences Inc., Richmond, USA.
authors.forename: Victoria
authors.initials: V
authors.lastname: Chu
authors.affiliation: Sangamo BioSciences Inc., Richmond, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Paschon
authors.affiliation: Sangamo BioSciences Inc., Richmond, USA.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Fred Hutchinson Cancer Research Center and University of Washington, Seattle, USA.
authors.forename: Jurgen
authors.initials: J
authors.lastname: Kuball
authors.affiliation: 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.
authors.forename: Barbara
authors.initials: B
authors.lastname: Camisa
authors.affiliation: 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.
authors.forename: Attilio
authors.initials: A
authors.lastname: Bondanza
authors.affiliation: 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.
authors.forename: Giulia
authors.initials: G
authors.lastname: Casorati
authors.affiliation: Department of Pathology, Leukemia Unit, San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Maurilio
authors.initials: M
authors.lastname: Ponzoni
authors.affiliation: Hematology Clinical Unit, Division of Regenerative Medicine, Gene Therapy and Stem Cells, San Raffaele Scientific Institute, Milan, Italy.|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.
authors.forename: Fabio
authors.initials: F
authors.lastname: Ciceri
authors.affiliation: Vita Salute San Raffaele University, Milan, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Bordignon
authors.affiliation: Fred Hutchinson Cancer Research Center and University of Washington, Seattle, USA.
authors.forename: Philip D
authors.initials: PD
authors.lastname: Greenberg
authors.affiliation: Sangamo BioSciences Inc., Richmond, USA.
authors.forename: Michael C
authors.initials: MC
authors.lastname: Holmes
authors.affiliation: Sangamo BioSciences Inc., Richmond, USA.
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy, Division of Regenerative Medicine, Gene Therapy and Stem Cells, San Raffaele Scientific Institute, Milan, Italy.|Vita Salute San Raffaele University, Milan, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Naldini
authors.affiliation: 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.|Vita Salute San Raffaele University, Milan, Italy.|Hematology Clinical Unit, Division of Regenerative Medicine, Gene Therapy and Stem Cells, San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Bonini
mesh_terms: D001483:Base Sequence; D015252:Deoxyribonucleases, Type II Site-Specific / Q000494:pharmacology*; D018014:Gene Transfer Techniques; D006801:Humans; D019169:Jurkat Cells; D016086:Lentivirus / Q000235:genetics*; D007938:Leukemia / Q000276:immunology*; D008969:Molecular Sequence Data; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D037182:T-Cell Antigen Receptor Specificity; D013601:T-Lymphocytes / Q000276:immunology; D025721:WT1 Proteins / Q000235:genetics; D016335:Zinc Fingers
chemical_list: D011948:Receptors, Antigen, T-Cell; D025721:WT1 Proteins; D015252:Deoxyribonucleases, Type II Site-Specific