Adaptable and scalable electroporation for cellular therapy

项目摘要 细胞疗法已经证明了治疗各种遗传性和获得性疾病的潜力。 疾病，但第一代批准的细胞疗法依赖于病毒载体的细胞 重新编程病毒载体被认为是发展这些药物的瓶颈。 治疗和其他递送方法，如电穿孔，正在临床上研究。到 为了解决现有电穿孔技术的局限性，CyteQuest正在开发一种简单， 可扩展的电穿孔平台，用于优化转染参数并递送货物 以高通量有效地和可再现地进行。CyteQuest的平台由一个平面流 具有薄板几何形状的芯片，其确保细胞经受均匀的电场， 可重复的电穿孔。流动池的几何形状允许无缝缩放到高- 用于临床规模递送的体积系统。本提案的主要目标是：（1） 展示临床相关货物的高效递送，包括CRISPR/Cas9 核糖核蛋白复合物与原代人T细胞的结合，以及（2）构建原型大- 体积电穿孔流动系统与所需的体积吞吐量兼容， 临床环境中的细胞疗法。CyteQuest将证明优化的 小体积系统的电穿孔参数可以直接用于大体积系统中， 该系统的生物分子递送效率和细胞活力的变化小于5%。的 高容量电穿孔流动系统可以在成本有效的方法中制造 和健壮。CyteQuest的最终目标是将这个新平台与 CRISPR/Cas9技术作为过继细胞疗法的治疗应用。

Project Summary Cellular therapies have demonstrated the potential to treat a variety of inherited and acquired diseases, but the first generation of approved cell therapies rely on viral vectors for cellular reprogramming. Viral vectors are recognized as a bottleneck in the development of these therapies and other delivery methods, like electroporation, are being clinically investigated. To address limitations in existing electroporation technology, CyteQuest is developing a simple, scalable electroporation platform to optimize transfection parameters and deliver cargo efficiently and reproducibly at high throughput. CyteQuest’s platform consists of a planar flow chip with a thin slab geometry that ensures that cells are subject to a uniform electrical field for reproducible electroporation. The flow cell geometry allows for seamless scaling to a high- volume system for delivery at clinical scale. The key objectives of this proposal are to: (1) demonstrate high-efficiency delivery of clinically relevant cargo including CRISPR/Cas9 ribonucleoprotein complexes to primary human T cells, and (2) construct a prototype large- volume electroporation flow system compatible with the volumetric throughput required for cellular therapy in a clinical setting. CyteQuest will demonstrate that the optimized electroporation parameters of the small volume system can be used directly in the high-volume system with less than 5% variation in the biomolecular delivery efficiency and cell viability. The high-volume electroporation flow system can be manufactured in a process that is cost-effective and robust. CyteQuest ultimately aims to use this novel platform in conjunction with CRISPR/Cas9 technology as a therapeutic application for adoptive cell therapy.