title: Immunological quality and performance of tumor vessel-targeting CAR-T cells prepared by mRNA-EP for clinical research.
abstract: We previously reported that tumor vessel-redirected T cells, which were genetically engineered with chimeric antigen receptor (CAR) specific for vascular endothelial growth factor receptor 2 (VEGFR2), demonstrated significant antitumor effects in various murine solid tumor models. In the present study, we prepared anti-VEGFR2 CAR-T cells by CAR-coding mRNA electroporation (mRNA-EP) and analyzed their immunological characteristics and functions for use in clinical research. The expression of anti-VEGFR2 CAR on murine and human T cells was detected with approximately 100% efficiency for a few days, after peaking 6-12 hours after mRNA-EP. Triple transfer of murine anti-VEGFR2 CAR-T cells into B16BL6 tumor-bearing mice demonstrated an antitumor effect comparable to that for the single transfer of CAR-T cells engineered with retroviral vector. The mRNA-EP did not cause any damage or defects to human T-cell characteristics, as determined by viability, growth, and phenotypic parameters. Additionally, two kinds of human anti-VEGFR2 CAR-T cells, which expressed different CAR construction, differentiated to effector phase with cytokine secretion and cytotoxic activity in antigen-specific manner. These results indicate that our anti-VEGFR2 CAR-T cells prepared by mRNA-EP have the potential in terms of quality and performance to offer the prospect of safety and efficacy in clinical research as cellular medicine.
authors.affiliation: Laboratory of Biotechnology and Therapeutics, Graduate School of Pharmaceutical Sciences, Osaka University , Osaka, Japan.
authors.forename: Kanako
authors.initials: K
authors.lastname: Inoo
authors.affiliation: Laboratory of Biotechnology and Therapeutics, Graduate School of Pharmaceutical Sciences, Osaka University , Osaka, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Inagaki
authors.affiliation: Laboratory of Biotechnology and Therapeutics, Graduate School of Pharmaceutical Sciences, Osaka University , Osaka, Japan.
authors.forename: Kento
authors.initials: K
authors.lastname: Fujiwara
authors.affiliation: MEDINET Co., Ltd. , Kanagawa, Japan.
authors.forename: Shigemi
authors.initials: S
authors.lastname: Sasawatari
authors.affiliation: MEDINET Co., Ltd., Kanagawa, Japan; Seta Clinic Group, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kamigaki
authors.affiliation: Laboratory of Biotechnology and Therapeutics, Graduate School of Pharmaceutical Sciences, Osaka University , Osaka, Japan.
authors.forename: Shinsaku
authors.initials: S
authors.lastname: Nakagawa
authors.affiliation: Laboratory of Biotechnology and Therapeutics, Graduate School of Pharmaceutical Sciences, Osaka University , Osaka, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Okada