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

摘要 用于将基因组修饰分子递送至细胞用于细胞疗法的两种主要方法是病毒、免疫球蛋白和免疫球蛋白。 转导和电穿孔转染具有阻碍细胞的增殖的过程和技术限制。 开发所有人都能使用的尖端细胞疗法。虽然病毒载体导致商品的高成本 由于与制造相关的重大监管和健康风险，电穿孔导致不健康的 具有低活力和脱靶表型变化的细胞。CellFE的微流体VECT技术使细胞 具有高转染效率、对细胞的低脱靶变化和扩增的细胞的转染 加工范围。 该项目旨在展示第一个商业化的细胞转染平台， CD 34+造血干/祖细胞（HSPC）的遗传修饰，以开发遗传和 感染性疾病，包括β地中海贫血、镰状细胞病、血友病和HIV。可扩展性意味着 在细胞工程的研究和开发阶段使用的工艺参数将立即适用于 临床规模（处理细胞数量增加300倍）。更简单的 制造规模扩大意味着更快的临床测试路径，将是一个重要的价值主张 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， 用于高通量的按比例放大器件通道，（2）将器件制造从PDMS转移到注塑成型 塑料，（3）以临床处理速率在实验室上展示治疗基因的遗传修饰，以及 (4)开发alpha原型系统，在我们的制药合作伙伴的实验室进行测试，以产生付费试点 问题研究这些都是SBIR二期项目的关键技术目标，也是CellFE的主要里程碑 为了执行其开发、营销和销售细胞转染的商业计划， 细胞治疗制造平台，具有降低制造开发风险和降低成本的前景 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.