Microfluidic CAR-T Cell Processing Device

微流控CAR-T细胞处理装置

基本信息

批准号：
9929262
负责人：
CURT I CIVIN
金额：
$ 92.44万
依托单位：
GPB SCIENTIFIC, INC.
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-04-17 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9929262
关键词：
Aliquot Autologous B-Cell Acute Lymphoblastic Leukemia Blood Blood Cells Blood Platelets Businesses CAR T cell therapy Cell Separation Cell Therapy Cell physiology Cells Cellular immunotherapy Centrifugation Childhood Clinical Clinical Trials Collaborations Cycloparaffins Device or Instrument Development Devices Doctor of Philosophy Engineering Erythrocytes FDA approved Geometry Goals Hand Harvest Hematologic Neoplasms Hematopoietic stem cells Human Immunophenotyping Immunotherapy Injections Lateral Leukapheresis Leukocytes Liquid substance Malignant Neoplasms Manufactured Materials Methods Microfluidic Microchips Microfluidics Molds Output Patients Phase Polymers Population Preparation Procedures Process Production Reagent Recovery Refractory Relapse Running Sampling Silicon Site Small Business Technology Transfer Research Solid Sterility Suspensions T-Lymphocyte Technology Testing Therapeutic Time Translating Work anticancer research base cancer cell cell injury cell type cellular targeting chimeric antigen receptor T cells cost cost effective design improved leukemia microchip novel operation performance tests personalized medicine preservation product development prototype scale up young adult

项目摘要

ABSTRACT The goal of this Fast-Track STTR project is to develop a Deterministic Lateral Displacement (DLD) microfluidic device that can enrich white blood cells (WBCs) from a typical leukapheresis unit in 1 hr, for use in manufacturing cancer cellular immunotherapy. Chimeric antigen receptor T cell (CAR-T) therapy has been recommended for FDA approval to treat relapsed or refractory pediatric and young adult patients with B-cell acute lymphoblastic leukemia. There is a critical need for cost-effective automated methods to improve the efficiency and yield of large-scale enrichment of WBCs for use in manufacturing CAR-T and other cellular therapies. GPB is a pioneer in developing novel DLD microchips to process blood cells for cell analysis (19,26). GPB now proposes to develop, evaluate and commercialize a compact device in which an entire leukapheresis unit (up to 5x1010 WBCs in up to 300 ml) can be processed in a “Leuko-stack” of disposable single-use multi-channel DLD chips to produce in 1 hr a washed cell suspension that is enriched in WBCs and depleted of red blood cells (RBCs) and platelets (PLT). In Phase I, Aim 1 is to increase cell throughput through the current prototype chips by: 1) optimizing DLD chip design and operation to increase flow rate; 2) increasing throughput by stacking plastic chips and running them in parallel (“Leuko-stacks”); and 3) translating chip production to high-volume manufacturing material such as Cyclic Olefin Polymer (COP). Final Phase I milestones to proceed to Phase II are: 1) final chip design with a flow rate of at least 25 mL/hr via a single chip, at least 70% recovery of viable WBCs and immunophenotype- defined T-lymphocytes, and ability to process cells for 1 hr without clogging; 2) Leuko-stack of at least 6 chips run in parallel, with the same output as in #1; 3) combined increases in throughput via #1 and #2 sufficient to process a 300 ml leukapheresis unit in 1 hr; 4) confirmation that the chips can be produced from COP. In Phase II, Aim 2 is to build final prototype COP plastic chip-based microfluidic device capable of processing a leukapheresis sample at 300 mL/hr. Aim 3 is to test performance of prototypes from Aim 2 with leukapheresis aliquots and then full-size human leukapheresis samples. The final milestone of this project is to produce a set of commercial prototype Leuko-stacks that can process an entire 300-ml leukapheresis unit in 1 hr with at least 70% WBC and T-lymphocyte recovery, at least 90% depletion of RBCs, at least 80% depletion of PLTs, and at least 70% recovery of T-cell expansion capacity (as compared with the input samples) in significantly more than 50% of samples tested at 2 sites. The GPB Leuko-stack platform will preserve the advantages of DLD microfluidic cell processing over current methods, while massively increasing throughput rate and cell processing capacity, thus transitioning from analytic- to preparative-scale WBC enrichment for subsequent manufacture of CAR-T and other cell therapies.

抽象的该快速通道 STTR 项目的目标是开发确定性横向位移 (DLD) 微流体装置可以在 1 小时内从典型的白细胞分离装置中富集白细胞 (WBC)，用于制造癌症细胞免疫疗法。已建议 FDA 批准用于治疗复发或难治性儿童和年轻成年患者 B 细胞急性淋巴细胞白血病迫切需要具有成本效益的自动化方法来改善。用于制造 CAR-T 和其他细胞的大规模富集 WBC 的效率和产量疗法。 GPB 是开发新型 DLD 微芯片以处理血细胞进行细胞分析的先驱 (19,26)。提议开发、评估和商业化一种紧凑型设备，其中整个白细胞分离装置（最多最多 300 毫升中的 5x1010 WBC）可在一次性多通道的“Leuko-stack”中进行处理 DLD 芯片可在 1 小时内产生富含白细胞并去除红血的洗涤细胞悬浮液细胞 (RBC) 和血小板 (PLT)。在第一阶段，目标 1 是通过以下方式提高当前原型芯片的单元吞吐量：1) 优化 DLD 芯片提高流量的设计和操作；2）通过堆叠塑料碎片并运行它们来提高吞吐量并行（“Leuko-stacks”）；以及 3）将芯片生产转化为大批量制造材料，例如环烯烃聚合物 (COP)。进入第二阶段的最终第一阶段里程碑是：1) 最终芯片设计通过单个芯片的流速至少为 25 mL/hr，活白细胞和免疫表型的回收率至少为 70% 明确的 T 淋巴细胞，以及处理细胞 1 小时而不会堵塞的能力；2) 至少 6 个芯片的白细胞堆栈；并行运行，输出与 #1 相同；3) 通过 #1 和 #2 增加组合吞吐量，足以在 1 小时内处理 300 ml 白细胞分离装置；4) 确认芯片可以由 COP 生产。在第二阶段，目标 2 是构建基于 COP 塑料芯片的最终原型微流体装置，能够处理以 300 mL/hr 的速度进行白细胞去除术样品目标 3 是测试目标 2 中白细胞去除术原型的性能。该项目的最后一个里程碑是生产一个等分试样，然后是全尺寸的人类白细胞分离术样本。一套商业原型白细胞堆栈，可在 1 小时内处理整个 300 毫升白细胞分离装置白细胞和 T 淋巴细胞恢复至少 70%，红细胞至少消耗 90%，至少 80% PLT 耗尽，T 细胞扩增能力至少恢复 70%（与输入相比）在 2 个地点测试的样品中，超过 50% 的样品）。 GPB Leuko-stack 平台将保留 DLD 微流控细胞处理相对于当前技术的优势方法，同时大幅提高吞吐率和细胞处理能力，从而从分析级到制备级白细胞富集，用于后续 CAR-T 和其他细胞疗法的生产。