Enabling pediatric leukapheresis with high-throughput microfluidic technology

利用高通量微流体技术实现儿科白细胞分离术

基本信息

批准号：
10604360
负责人：
Sergey S Shevkoplyas
金额：
$ 38.96万
依托单位：
UNIVERSITY OF HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10604360
关键词：
Address Adult Affect Allergic Reaction Anemia Anesthesia procedures Animal Model Animals Anticoagulation Back Blood Blood Cell Count Blood Cells Blood Component Removal Blood Platelets Blood Volume Blood specimen Carotid Arteries Catheters Cell Separation Cell Size Cell Therapy Cells Centrifugation Cessation of life Child Child Welfare Childhood Circulation Clinical Collagen Collection Complete Blood Count Data Development Device Designs Devices Dyes Ensure Erythrocytes Excision Family suidae Filtration Flow Cytometry Geometry Glycoproteins Health Hematocrit procedure Hematology Hemoglobin Hypocalcemia Hypotension Immune System Diseases In Vitro Incidence Individual Infant Infection Inflammatory Interleukin-1 Internal jugular vein structure Kinetics Knowledge Leukapheresis Leukocytes Lymphocyte Macrophage-1 Antigen Measures Medical Methods Microfluidic Microchips Mononuclear Morphology P-Selectin Patients Performance Peripheral Plasma Platelet Activation Potassium Pre-Clinical Model Procedures Process Property Pump Recovery Research Resistance Sampling Suspensions TNF gene Technology Testing Thrombosis Validation Vulnerable Populations Weight White Blood Cell Count procedure Whole Blood Work cell injury clinical risk cytokine density design experience healthy volunteer human subject improved in vivo invention iterative design manufacture microfluidic technology miniaturize mouse model neonate novel operation pediatric patients performance tests prototype research clinical testing

项目摘要

PROJECT SUMMARY Leukapheresis is a specialized medical procedure during which patient's whole blood (WB) is passed through the extracorporeal circuit of an apheresis machine, which extracts white blood cells (WBCs) from WB, and returns red blood cells (RBCs) and platelets (PLTs) back to the patient. The separation of WBCs from patient's blood via leukapheresis is the key initial step for an increasing number of highly effective cell-based treatments for some of the most devastating hematologic and immune system disorders affecting millions of adults and children worldwide. Currently, leukapheresis is performed using centrifugation-based apheresis machines, which have a substantial extracorporeal volume (ECV). Although well-tolerated by most adults and older children, leukapheresis in young children weighing less than about 10 kg (or 22 lbs) is technically challenging and clinically risky. Because ECV represents a particularly large fraction of their total blood volume (TBV), these vulnerable patients experience a significantly higher incidence of hypotension, symptomatic hypocalcemia, allergic reactions, catheter-related thrombosis, infections, severe anemia and even death. There is currently no practical alternative to adult-size apheresis machines for performing leukapheresis in neonates and low-weight infants. To address this significant limitation, we will develop and validate novel high-throughput microfluidic devices with very low void volume, capable of separating WBCs from WB with volumetric throughput and efficiency sufficiently high to ultimately enable centrifugation-free, low-ECV leukapheresis. These devices will utilize new cell separation technology (`controlled incremental filtration', or CIF) which we have previously applied to separating WBCs from concentrated blood cell suspensions with high efficiency, minimal RBC and PLT loss, and at flow rates on par with conventional leukapheresis. Here we will apply this approach to separating WBCs directly from WB by completing three complementary aims with scope ranging from iterative design optimization and validation work, to testing the performance of the CIF-based leukapheresis devices in an animal model. First, we will optimize the CIF design parameters to maximize WBC separation, while minimizing RBC/PLT losses and device fluidic resistance when processing WB. Second, we will multiplex individual CIF device modules into full- scale device prototypes, optimize their operation in the recirculation regime, and validate their ability to process large volumes of WB in vitro. Third, we will comprehensively evaluate device performance and the effect of CIF- based processing on blood cell properties in a mouse model. Detailed blood cell counts and markers of cell activation and damage will be measured throughout the project to aid the iterative design process and to validate the use of CIF technology for leukapheresis. By completing this research, we will develop functional device prototypes and generate pivotal data to support further testing in a pre-clinical model (porcine), before finalizing the device design for manufacturing from thermoplastic and clinical testing in human subjects.

项目概要白细胞分离术是一种特殊的医疗程序，在此过程中，患者的全血 (WB) 会被通过单采机的体外回路，从 WB 中提取白细胞 (WBC)，然后返回红细胞 (RBC) 和血小板 (PLT) 返回患者体内。从患者血液中分离白细胞通过白细胞去除术是越来越多高效的基于细胞的治疗的关键初始步骤影响数百万成人和儿童的一些最具破坏性的血液和免疫系统疾病全世界。目前，白细胞分离术是使用基于离心的分离机进行的，该机器具有大量的体外体积（ECV）。尽管大多数成人和年龄较大的儿童都具有良好的耐受性，对体重小于 10 公斤（或 22 磅）的幼儿进行白细胞分离术在技术上和临床上都具有挑战性有风险。由于 ECV 占其总血量 (TBV) 的很大一部分，因此这些弱势群体患者出现低血压、症状性低钙血症、过敏的发生率明显更高反应、导管相关血栓形成、感染、严重贫血甚至死亡。目前还没有实用的成人尺寸的单采机的替代品，用于对新生儿和低体重婴儿进行白细胞分离术。为了解决这一重大限制，我们将开发和验证新型高通量微流体装置极低的空隙体积，能够以足够的体积通量和效率将 WBC 与 WB 分离高最终实现免离心、低 ECV 白细胞分离术。这些设备将利用新的电池我们之前已将其应用于分离的分离技术（“控制增量过滤”或 CIF）从浓缩血细胞悬浮液中提取白细胞，效率高，RBC 和 PLT 损失极少，并且处于流动状态率与传统白细胞分离术相当。在这里，我们将应用这种方法直接从 WB 通过完成三个互补的目标，范围从迭代设计优化到验证工作，以在动物模型中测试基于 CIF 的白细胞分离装置的性能。第一的，我们将优化 CIF 设计参数，以最大限度地提高 WBC 分离，同时最大限度地减少 RBC/PLT 损失和处理 WB 时的设备流体阻力。其次，我们将把各个 CIF 设备模块复用到全缩放设备原型，优化其在再循环状态下的操作，并验证其处理能力体外大量WB。第三，我们将综合评估设备性能和CIF-的效果基于小鼠模型中血细胞特性的处理。详细的血细胞计数和细胞标记将在整个项目中测量激活和损坏，以帮助迭代设计过程并验证使用 CIF 技术进行白细胞分离术。通过完成这项研究，我们将开发功能器件在最终确定之前，制作原型并生成关键数据以支持临床前模型（猪）中的进一步测试用于热塑性塑料制造和人体临床测试的设备设计。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Author Correction: A high-throughput microfluidic approach for 1000-fold leukocyte reduction of platelet-rich plasma.

DOI：
10.1038/s41598-022-18449-5
发表时间：
2022-08-29
期刊：
SCIENTIFIC REPORTS
影响因子：
4.6
作者：
Xia, Hui;Strachan, Briony C.;Gifford, Sean C.;Shevkoplyas, Sergey S.
通讯作者：
Shevkoplyas, Sergey S.

A high-throughput microfluidic device based on controlled incremental filtration to enable centrifugation-free, low extracorporeal volume leukapheresis.

DOI：
10.1038/s41598-022-16748-5
发表时间：
2022-08-13
期刊：
SCIENTIFIC REPORTS
影响因子：
4.6
作者：
Lezzar, Dalia L.;Lam, Fong W.;Huerta, Ravin;Mukhamedshin, Anton;Lu, Madeleine;Shevkoplyas, Sergey S.
通讯作者：
Shevkoplyas, Sergey S.

Rapid, label-free enrichment of lymphocytes in a closed system using a flow-through microfluidic device.