Microfluidic vortex shedding: a low-cost, high efficiency method for genetic modification to support cell engineering for cell-based immunotherapies

个性化的基因修饰细胞免疫疗法是存在的，但制造尚未优化，以增加这些拯救生命的疗法在有需要的患者中的广泛可获得性。这项建议的重点是通过微流控涡流剥离(μVS)的持续发展来满足这一需求，微流控涡流剥离是一种安全和快速的方法，可以对患者来源的免疫细胞进行基因修饰。这项提案的长期目标是将μVS整合到基于细胞的癌症免疫疗法的生产流程中。这份合同提案的目的是证明μVS技术在产生具有代表性的嵌合抗原受体T细胞(CAR-T)和T细胞受体T细胞(TCR-T)方面的可行性。研究和开发的目标是：(1)展示μVS用临床相关的CAR和TCRs构建的原代T细胞的技术性能，以及(2)在基于细胞的检测中证明μVS产生的转染性T细胞的功能性和安全性。在这些目标成功完成之前，CAR-T和TCR-T细胞将使用患者来源的T细胞进行工程设计，并将展示商业规模的加工和浓缩足够的转基因活细胞用于临床应用。

Personalized gene-modified cell immunotherapies exist, but manufacturing has yet to be optimized to increase the broad availability of these life-saving therapies to patients in need. This proposal is focused on meeting this need with the continued development of microfluidic vortex shedding (μVS), a safe and rapid approach to genetically modify patient-derived immune cells. The long term objective of this proposal is to integrate μVS into the manufacturing workflow of cell-based cancer immunotherapies. The goal of this contract proposal to demonstrate the feasibility of μVS technology in generating representative Chimeric Antigen Receptor T cells (CAR-T) and T Cell Receptor T cells (TCR-T) cells. The research and development objectives are to: (1) demonstrate the technical performance of μVS transfection of primary T cells with clinically relevant CAR and TCR constructs, and (2) demonstrate the functionality and safety of transfected T cells generated by μVS in cell based assays. Pending the successful completion of these objectives, CAR-T and TCR-T cells will be engineered using patient-derived T cells, and commercial-scale processing and enrichment of sufficient genetically modified viable cells for clinical applications will be demonstrated.