Efficient and scalable microfluidic gene delivery to hematopoietic stem cells to treat blood diseases

摘要 将基因组修饰分子运送到细胞进行细胞治疗的两种主要方法是病毒 转导和电穿孔转基因，具有工艺和技术限制，阻碍了 开发所有人都能获得的尖端细胞疗法。虽然病毒载体会导致高昂的商品成本 由于与制造相关的重大监管和健康风险，电穿孔会导致不健康 存活率低和非靶向表型改变的细胞。CellFE的微流控Vect技术使CELL 具有高转染率、低偏离靶向改变的细胞和具有扩增细胞的转染体 加工范围。 该项目旨在展示为可扩展的体外实验而设计的第一个商业细胞转导平台 CD34+造血干/祖细胞(HSPC)的基因修饰 传染性疾病，包括贝塔地中海贫血、镰状细胞病、血友病和艾滋病毒。可伸缩性意味着 细胞工程研究和开发阶段使用的工艺参数将立即应用于 临床规模(处理细胞数量增加300倍)，只需更换一个芯片即可。一个更简单的 制造规模扩大意味着更快地进行临床测试，这将是 技术为了实现这些目标，CellFE将追求以下目标：(1)确定最佳设计，以 用于高通量的放大器件通道，(2)从PDMS到注塑的转移器件制造 塑料，(3)以临床处理速度在工作台上演示治疗基因的遗传修改，以及 (4)开发阿尔法原型系统，以便在我们的制药合作伙伴的实验室中进行测试，以生成付费试点 学习。这些是SBIR第二阶段项目的主要技术目标，也是CellFE的主要里程碑 需要完成以执行其开发、营销和销售细胞转基因的商业计划 具有降低制造发展风险和降低成本前景的细胞治疗制造平台 商品与当前交付技术的对比。

Abstract The two dominant methods for delivering genome modifying molecules to cells for cell therapies, viral transduction and electroporation transfection, have process and technical limitations that hamper the development of cutting-edge cell therapies accessible to all. While viral vectors result in a high cost of goods with significant regulatory and health risks associated with manufacturing, electroporation results in unhealthy cells with low viability and off-target phenotype changes. CellFE’s microfluidic VECT technology enables cell transfections with high transfection efficiency, low off-target changes to the cell, and with an expanded cell processing range. This project aims to demonstrate the first commercial cell transfection platform designed for scalable ex vivo genetic modification of CD34+ hematopoietic stem/progenitor cells (HSPC) to develop treatments of genetic and infectious diseases including beta thalassemia, sickle cell disease, hemophilia, and HIV. Scalability means that process parameters used in the research and development phase of cell engineering will immediately apply to clinical scale (300-fold increase in number of processed cells) by simply exchanging a chip. A simpler manufacturing scale-up means a faster path to clinical testing and will be a significant value proposition for the technology. To achieve these goals, CellFE will pursue the following aims: (1) determine the optimal design to scaleup device channels for high-throughput, (2) transfer device fabrication from PDMS to injection molded plastic, (3) demonstrate genetic modification of a therapeutic gene at bench-top at a clinical processing rate, and (4) develop alpha prototype systems for testing in the labs of our pharmaceutical partners to generate paid pilot studies. These are the key technical aims of the SBIR Phase II project and are also the main milestones CellFE needs to accomplish in order to execute its business plan of developing, marketing, and selling cell transfection platforms for cell therapy manufacturing with the prospect to de-risk manufacturing development and reduce cost of goods versus current delivery techniques.