title: The Tol2 transposon system mediates the genetic engineering of T-cells with CD19-specific chimeric antigen receptors for B-cell malignancies.
abstract: Engineered T-cell therapy using a CD19-specific chimeric antigen receptor (CD19-CAR) is a promising strategy for the treatment of advanced B-cell malignancies. Gene transfer of CARs to T-cells has widely relied on retroviral vectors, but transposon-based gene transfer has recently emerged as a suitable nonviral method to mediate stable transgene expression. The advantages of transposon vectors compared with viral vectors include their simplicity and cost-effectiveness. We used the Tol2 transposon system to stably transfer CD19-CAR into human T-cells. Normal human peripheral blood lymphocytes were co-nucleofected with the Tol2 transposon donor plasmid carrying CD19-CAR and the transposase expression plasmid and were selectively propagated on NIH3T3 cells expressing human CD19. Expanded CD3(+) T-cells with stable and high-level transgene expression (~95%) produced interferon-γ upon stimulation with CD19 and specifically lysed Raji cells, a CD19(+) human B-cell lymphoma cell line. Adoptive transfer of these T-cells suppressed tumor progression in Raji tumor-bearing Rag2(-/-)γc(-/-) immunodeficient mice compared with control mice. These results demonstrate that the Tol2 transposon system could be used to express CD19-CAR in genetically engineered T-cells for the treatment of refractory B-cell malignancies.
authors.affiliation: 1] Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan [2] Division of Immuno-Gene and Cell Therapy (Takara Bio), Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Tsukahara
authors.affiliation: Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Iwase
authors.affiliation: Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Shizuoka, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Kawakami
authors.affiliation: Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Shizuoka, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Iwasaki
authors.affiliation: 1] Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan [2] Division of Hematology, Department of Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: C
authors.initials: C
authors.lastname: Yamamoto
authors.affiliation: 1] Division of Immuno-Gene and Cell Therapy (Takara Bio), Jichi Medical University, Shimotsuke, Tochigi, Japan [2] Division of Hematology, Department of Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ohmine
authors.affiliation: 1] Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan [2] Division of Immuno-Gene and Cell Therapy (Takara Bio), Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Uchibori
authors.affiliation: Division of Immuno-Gene and Cell Therapy (Takara Bio), Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Teruya
authors.affiliation: Division of Immuno-Gene and Cell Therapy (Takara Bio), Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Ido
authors.affiliation: 1] Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan [2] Department of Obstetrics and Gynecology, Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Saga
authors.affiliation: Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Urabe
authors.affiliation: Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Mizukami
authors.affiliation: Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Kume
authors.affiliation: Human Gene Sciences Center, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Nakamura
authors.affiliation: Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
authors.forename: R
authors.initials: R
authors.lastname: Brentjens
authors.affiliation: 1] Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan [2] Division of Immuno-Gene and Cell Therapy (Takara Bio), Jichi Medical University, Shimotsuke, Tochigi, Japan [3] Division of Hematology, Department of Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ozawa
mesh_terms: D000818:Animals; D018941:Antigens, CD19 / Q000276:immunology*; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D004251:DNA Transposable Elements; D005818:Genetic Engineering; D015316:Genetic Therapy; D006801:Humans; D016219:Immunotherapy, Adoptive; D016393:Lymphoma, B-Cell / Q000628:therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D009368:Neoplasm Transplantation; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D018941:Antigens, CD19; D004251:DNA Transposable Elements; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins