title: Mesothelin-targeting chimeric antigen receptor-modified T cells by piggyBac transposon system suppress the growth of bile duct carcinoma.
abstract: Chimeric antigen receptor modified T cell-based immunotherapy is revolutionizing the field of cancer treatment. However, its potential in treating bile duct carcinoma has not been fully explored. Herein, we developed the second-generation mesothelin-targeting chimeric antigen receptor-modified T cells with the 4-1BB co-stimulatory module by the piggyBac transposon system. Mesothelin-targeting chimeric antigen receptor was expressed by 66.0% of mesothelin-targeting chimeric antigen receptor-modified T cells post electrophoretic transfection and stimulation with K562-meso cells; the expressions of activation markers were tested by flow cytometry assay and showed greater activation of mesothelin-targeting chimeric antigen receptor-modified T cells than control T cells (CD107α: 71.9% vs 48.6%; CD27: 92.1% vs 61.8%; CD137: 55.5% vs 8.4%; CD28: 98.0% vs 82.1%; CD134: 37.5% vs 10.4%). Furthermore, mesothelin-targeting chimeric antigen receptor-modified T cells exerted cytotoxicity toward mesothelin-expressing EH-CA1b and EH-CA1a cells in an effector-to-target ratio-dependent manner, while leaving mesothelin-negative GSC-SD and EH-GB1 cells and normal liver L02 cells almost unharmed. Mesothelin-targeting chimeric antigen receptor-modified T cells secreted cytokines at higher levels when co-cultured with mesothelin-positive EH-CA1a and EH-CA1b cells than with mesothelin-negative GSC-SD and EH-GB1 cells. Enhanced cytotoxicity and cytokine secretion of mesothelin-targeting chimeric antigen receptor-modified T cells compared to control T cells were also observed when co-cultured with 293-meso cells (interferon γ: 85.1% ± 1.47% vs 8.3% ± 2.50%, p = 0.000; tumor necrosis factor α: 90.9% ± 4.67% vs 18.5% ± 3.62%, p = 0.0004; interleukin 2: 60.8% ± 2.00% vs 15.6% ± 2.06%, p = 0.002; interleukin 6: 6.4% ± 2.95% vs 1.7% ± 0.63%, p = 0.055). In addition, mesothelin-targeting chimeric antigen receptor-modified T cells showed greater inhibitory and proliferative capability than control T cells within EH-CA1a cell xenografts. This study shows the potential of mesothelin-targeting chimeric antigen receptor-modified T cells in treating bile duct carcinoma.
authors.affiliation: 1 Xinyuan Institute of Medicine and Biotechnology, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China.
authors.forename: Jie-Ying
authors.initials: JY
authors.lastname: Xu
authors.affiliation: 2 Laboratory of Gene and Viral Therapy, Eastern Hepatobiliary Surgery Hospital, The Second Military Medical University of Chinese PLA, Shanghai, China.
authors.forename: Zhen-Long
authors.initials: ZL
authors.lastname: Ye
authors.affiliation: 1 Xinyuan Institute of Medicine and Biotechnology, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China.
authors.forename: Du-Qing
authors.initials: DQ
authors.lastname: Jiang
authors.affiliation: 1 Xinyuan Institute of Medicine and Biotechnology, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China.
authors.forename: Jiang-Chuan
authors.initials: JC
authors.lastname: He
authors.affiliation: 3 Department of Biotherapy, Eastern Hepatobiliary Surgery Hospital, The Second Military Medical University of Chinese PLA, Shanghai, China.
authors.forename: Yong-Mei
authors.initials: YM
authors.lastname: Ding
authors.affiliation: 2 Laboratory of Gene and Viral Therapy, Eastern Hepatobiliary Surgery Hospital, The Second Military Medical University of Chinese PLA, Shanghai, China.
authors.forename: Lin-Fang
authors.initials: LF
authors.lastname: Li
authors.affiliation: 2 Laboratory of Gene and Viral Therapy, Eastern Hepatobiliary Surgery Hospital, The Second Military Medical University of Chinese PLA, Shanghai, China.
authors.forename: Sai-Qun
authors.initials: SQ
authors.lastname: Lv
authors.affiliation: 2 Laboratory of Gene and Viral Therapy, Eastern Hepatobiliary Surgery Hospital, The Second Military Medical University of Chinese PLA, Shanghai, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: 2 Laboratory of Gene and Viral Therapy, Eastern Hepatobiliary Surgery Hospital, The Second Military Medical University of Chinese PLA, Shanghai, China.
authors.forename: Hua-Jun
authors.initials: HJ
authors.lastname: Jin
authors.affiliation: 1 Xinyuan Institute of Medicine and Biotechnology, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China.|2 Laboratory of Gene and Viral Therapy, Eastern Hepatobiliary Surgery Hospital, The Second Military Medical University of Chinese PLA, Shanghai, China.|3 Department of Biotherapy, Eastern Hepatobiliary Surgery Hospital, The Second Military Medical University of Chinese PLA, Shanghai, China.
authors.forename: Qi-Jun
authors.initials: QJ
authors.lastname: Qian
mesh_terms: D000818:Animals; D001650:Bile Duct Neoplasms / Q000473:pathology / Q000628:therapy*; D002478:Cells, Cultured; D004251:DNA Transposable Elements; D058851:GPI-Linked Proteins / Q000276:immunology*; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D000090204:Mesothelin; D051379:Mice; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D011993:Recombinant Fusion Proteins; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D004251:DNA Transposable Elements; D058851:GPI-Linked Proteins; C000715528:Msln protein, mouse; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D000090204:Mesothelin
keywords: Chimeric antigen receptor; bile duct carcinoma; immunotherapy; mesothelin; piggyBac