Regeneration of T cells from human-induced pluripotent stem cells for CAR-T cell medicated immunotherapy

作者全名："Chen, Yanyan; Huang, Pufeng; Niu, Mengda; Tian, Chuanhuizi; Zhang, Tingting; Peng, Zhiping"

作者地址："[Chen, Yanyan; Huang, Pufeng; Niu, Mengda; Tian, Chuanhuizi; Zhang, Tingting; Peng, Zhiping] Chongqing Med Univ, Sch Basic Med, Dept Radiat Med, Chongqing, Peoples R China"

通信作者："Peng, ZP (通讯作者)，Chongqing Med Univ, Sch Basic Med, Dept Radiat Med, Chongqing, Peoples R China."

来源：FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY

ESI学科分类：Multidisciplinary

WOS号：WOS:001013690800001

JCR分区：Q1

影响因子：4.3

年份：2023

卷号：11

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：immunotherapy; cell differentiation; human induced pluripotent stem cells; lymphomagenesis; hiPS-CAR-T cells

摘要："Background: Chimeric antigen receptor (CAR) T cell treatment involves in vitro production of T cells from patient blood with synthetic receptors specific to a cancer antigen. They circumvent the major histocompatibility complex to recognize the tumor antigen, reducing hematologic malignancy remission rates by 80%. Considering the efficacy of CAR-T treatment, the present work aimed at generating functional clusters of differentiation (CD)8 + T cells from human induced pluripotent stem cells (hiPSC) and to generate hiPS-CAR-T cells with high antigen-specific cytotoxicity.Methods: The Alkaline phosphatase assay and MycoEasy rapid mycoplasma detection kit was implemented for detection of hiPSCs and mycoplasma, respectively. The CD34(+) HSPCs were harvested in AggreWellTM 400 using a 37-micron reversible strainer. Likewise, the lymphoid progenitor and CD4(+)CD8(+) DP T cells were also harvested. The Cell Counting Kit-8 (CCK-8) assay was used to mark cytotoxicity and ELISA was used to detect IFN-? secretion. Further, flow cytometry and transwell chambers were used to assess cell cycle, and migration and invasion. Finally, the in vivo antitumor effects of the CAR-T cells were evaluated using experimental animals (mice).Results: Results revealed that a serum-free, feeder layer-free differentiation system significantly yielded hiPSC-based T cell immunotherapy with interleukin-2, interleukin-15, and activators at the differentiation stage to promote the maturation of these cells into human induced pluripotent stem (hiPS)-T cells. The infection of hiPSCs with the CD19 CAR lentivirus resulted in the production of the hiPSC-CAR-T cells. We validated the function of hiPS-CAR-T cells in vivo and in vitro experimentation which revealed no significant differences in cell morphology and function between hiPSC-derived hiPS-CAR-T cells and peripheral blood-derived CAR-T cells.Conclusion: This study developed a culture method that is efficient and clinically useful to make functional CD8(+) T cells from hiPSC and to get hiPS-CAR-T cells with high antigen-specific cytotoxicity that are not very different from CAR T cells found in peripheral blood. As a result, our findings may open the way for the clinical use of hiPSC to create functional CD8(+) T and hiPS-CAR-T cells cells for use in cell-based cancer therapy."

基金机构：Natural Science Foundation of China [81771871]

基金资助正文："Funding This work was supported by the Natural Science Foundation of China under grant number 81771871, the visualization and biological characterization of CAR-T lymphocytes constructed by CRISPR/Cas9-mediated genome editing and iPSCs directional differentiation."