Adaptable and scalable electroporation for cellular therapy

用于细胞治疗的适应性和可扩展的电穿孔

基本信息

批准号：
10545845
负责人：
HAROLD G CRAIGHEAD
金额：
$ 27.52万
依托单位：
CYTEQUEST, INC.
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10545845
关键词：
Address Adoptive Cell Transfers Benchmarking Budgets CRISPR/Cas technology Cell Survival Cell Therapy Cell physiology Cells Cellular biology Clinical Complex DNA DNA delivery Devices Disease Electroporation Ensure Environment Generations Genes Geometry Green Fluorescent Proteins Human Inherited Jurkat Cells Knock-out Liquid substance Messenger RNA Methods Modification Nucleic Acids Performance Phase Process Production Proteins RNA Reproducibility Research Ribonucleoproteins Sampling System T-Lymphocyte Technology Therapeutic Thinness Transfection Transposase Vaccination Validation Variant Viral Viral Vector chimeric antigen receptor chimeric antigen receptor T cells clinically relevant clinically significant cost effective design drug discovery electric field experience fluid flow knock-down mRNA Expression mRNA delivery novel plasmid DNA programmed cell death protein 1 prototype receptor stem cell therapy therapy development transgene expression transposon/insertion element trend

项目摘要

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.

项目概要细胞疗法已证明具有治疗多种遗传性和获得性遗传性疾病的潜力疾病，但第一代批准的细胞疗法依赖于病毒载体来治疗细胞病毒载体被认为是这些发展的瓶颈。疗法和其他递送方法（例如电穿孔）正在临床研究中。为了解决现有电穿孔技术的局限性，CyteQuest 正在开发一种简单的、可扩展的电穿孔平台，用于优化转染参数并输送货物 CyteQuest 的平台由平面流组成，可在高通量下高效且可重复地进行。芯片具有薄板几何形状，可确保细胞受到均匀的电场作用可重复的电穿孔几何形状允许无缝缩放至高通量。该提案的主要目标是：（1）展示临床相关货物（包括 CRISPR/Cas9）的高效递送核糖核蛋白复合物与原代人 T 细胞，以及 (2) 构建原型大体积电穿孔流系统与所需的体积通量兼容 CyteQuest 将在临床环境中证明细胞疗法的优化。小容量系统的电穿孔参数可直接用于大容量系统系统的生物分子递送效率和细胞活力的变化小于 5%。大容量电穿孔流路系统的制造过程具有成本效益 CyteQuest 最终目标是结合使用这个新颖的平台。 CRISPR/Cas9 技术作为过继细胞疗法的治疗应用。