High Efficiency Microfluidic Device for Large Scale CAR-T Cell Manufacturing

项目总结/摘要 本项目的目标是开发一种用于富集白色血细胞的高产量微流控装置 白细胞去除产品中的白细胞（WBC），用于制造CAR-T和其他癌症细胞 免疫疗法靶向CD 19的CAR-T细胞疗法非常有效，FDA批准用于治疗 晚期血液恶性肿瘤，目前有超过300项正在进行的临床试验使用CAR-T细胞， 针对各种癌症，包括实体瘤。大规模生产细胞免疫疗法， 包括CAR-T细胞疗法，由于每种疗法都是定制的， 由癌症患者自己的白细胞制造，使用白细胞分离术收集。因为癌症 患者通常具有低WBC计数和不一致的外周通路，他们的白细胞分离收获物 通常WBC数量有限，并且被红细胞（RBC）高度污染。有一个 对于用于CAR-T细胞制造的WBC源材料的高产量富集的迫切需要。CG 科学正在开发一种具有成本效益的设备，以高产量富集WBC，基于其 专利微流控“高效确定性分离（HEDS）”技术。技术 旨在（1）提供细胞癌症免疫治疗所必需的高WBC恢复，以及（2） 克服了阻碍其它微流体细胞分离的成本和生产量问题 成功商业化的技术。初步数据显示，HEDS能够 提供>95%的WBC回收率和>98%的RBC去除率，细胞活力损失不可检测- 显著优于现有的WBC富集系统。HEDS的独特配置可以 潜在地提供可扩展的处理量、抗堵塞性以及通过以下方式操作的能力： 重力进料为了进一步确定HEDS技术用于电池制造的可行性，本第一阶段 项目将集中在3个具体目标：（1）优化HEDS芯片设计和开发芯片包，以扩大规模 处理通量，（2）证明高WBC回收率并表征富集的WBC 流式细胞术和T细胞增殖试验，以及（3）显示塑料HEDS 使用软压纹的芯片可制造性。研究人员团队包括HEDS的发明者 技术，免疫学生物学家，塑料设备制造工程师和医生， 专攻细胞治疗和细胞制造这个项目的成功将导致高产量 WBC富集装置，将显著改善细胞免疫疗法的源材料， 使细胞制造过程更加稳健和可靠，并最终提供更有效的， 具有成本效益的靶向癌症治疗将使许多患者受益。

Project Summary/Abstract The goal of this project is to develop a high-yield microfluidic device for enriching white blood cells (WBCs) from leukapheresis products, for use in manufacturing CAR-T and other cancer cellular immunotherapies. CD19-targeted CAR-T cell therapy is highly effective and FDA-approved for treating advanced hematologic malignancies, and over 300 active clinical trials are currently using CAR-T cells to target various cancers, including solid tumors. Large-scale production of cellular immunotherapy, including CAR-T cell therapy, poses a unique challenge due to the fact that each therapy is custom- manufactured from the cancer patient’s own WBCs, collected using leukapheresis. Because cancer patients often have low WBC counts and inconsistent peripheral access, their leukapheresis harvests are frequently limited in WBC quantities and highly contaminated with red blood cells (RBCs). There is a critical need for high-yield enrichment of the WBC source material for CAR-T cell manufacturing. CG Scientific is developing a cost-effective device that enriches WBCs with high yields, based on its patented microfluidic “High Efficiency Deterministic Separation (HEDS)” technology. The technology aims to (1) provide high WBC recovery that is essential to cellular cancer immunotherapy, and (2) overcome the cost and throughput issues that have precluded other microfluidic cell separation technologies from successful commercialization. Preliminary data have shown that HEDS is capable of delivering >95% WBC recovery and >98% RBC removal, with undetectable loss of cell viability— significantly outperforming existing WBC enrichment systems. The unique configuration of HEDS can potentially provide scalable processing throughput, resistance to clogging, and capability to operate by gravity feed. To further establish the feasibility of HEDS technology for cell manufacturing, this Phase I project will focus on 3 Specific Aims: (1) optimize HEDS chip design and develop Chip Packs to scale up processing throughput, (2) demonstrate high WBC recovery yields and characterize the enriched WBCs and T cell subsets using flow cytometry and T cell proliferation assay, and (3) demonstrate plastic HEDS chip manufacturability using soft embossing. The investigator team includes the inventor of the HEDS technology, immunology biologists, plastic device manufacturing engineers, and medical doctors who specialize in cellular therapy and cell manufacturing. The success of this project will lead to a high-yield WBC enrichment device that will significantly improve the source material for cellular immunotherapies, make the cell manufacturing process more robust and reliable, and eventually provide more potent and cost-effective targeted cancer therapies that will benefit many patients.