Sytsma Brandon J, Allain Vincent, Bourke Struan, Faizee Fairuz, Fathi Mohsen, Berdeaux Rebecca, Ferreira Leonardo M R, Brewer W Jared, Li Lian, Pan Fong L, Rothrock Allison G, Nyberg William A, Li Zhongmei, Wilson Leah H, Eyquem Justin, Pawell Ryan S
Indee Labs, Berkeley, CA, USA.
Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA.
bioRxiv. 2024 Jul 13:2024.06.25.600671. doi: 10.1101/2024.06.25.600671.
Adoptive chimeric antigen receptor T-cell (CAR-T) therapy is transformative and approved for hematologic malignancies. It is also being developed for the treatment of solid tumors, autoimmune disorders, heart disease, and aging. Despite unprecedented clinical outcomes, CAR-T and other engineered cell therapies face a variety of manufacturing and safety challenges. Traditional methods, such as lentivirus transduction and electroporation, result in random integration or cause significant cellular damage, which can limit the safety and efficacy of engineered cell therapies. We present hydroporation as a gentle and effective alternative for intracellular delivery. Hydroporation resulted in 1.7- to 2-fold higher CAR-T yields compared to electroporation with superior cell viability and recovery. Hydroporated cells exhibited rapid proliferation, robust target cell lysis, and increased pro-inflammatory and regulatory cytokine secretion in addition to improved CAR-T yield by day 5 post-transfection. We demonstrate that scaled-up hydroporation can process 5 x 10 cells in less than 10 s, showcasing the platform as a viable solution for high-yield CAR-T manufacturing with the potential for improved therapeutic outcomes.
过继性嵌合抗原受体T细胞（CAR-T）疗法具有变革性，已被批准用于治疗血液系统恶性肿瘤。它也正在被开发用于治疗实体瘤、自身免疫性疾病、心脏病和衰老。尽管取得了前所未有的临床疗效，但CAR-T和其他工程细胞疗法面临着各种制造和安全挑战。传统方法，如慢病毒转导和电穿孔，会导致随机整合或造成显著的细胞损伤，这可能会限制工程细胞疗法的安全性和有效性。我们提出水穿孔法作为一种温和且有效的细胞内递送替代方法。与电穿孔相比，水穿孔法产生的CAR-T产量高出1.7至2倍，且细胞活力和恢复情况更佳。除了在转染后第5天提高CAR-T产量外，经水穿孔处理的细胞还表现出快速增殖、强大的靶细胞裂解能力以及促炎和调节性细胞因子分泌增加。我们证明，扩大规模的水穿孔法可在不到10秒的时间内处理5×10个细胞，这表明该平台是一种可行的高产CAR-T制造解决方案，具有改善治疗效果的潜力。
