Eliminating Mediators of Toxicity from Stored Blood

描述（由申请人提供）：在美国，每年有近1500万单位的红细胞（RBC）输注给约500万患者，RBC输注是住院患者最常用的处方疗法之一。近年来，大量的临床证据表明，危重病人的发病率和死亡率中有很大一部分是由于输注红细胞的毒性作用。大多数输血涉及在1-6 ℃下储存在抗凝剂-防腐剂溶液中长达6周的RBC。RBC的生物化学、机械和功能特性逐渐恶化-到允许储存结束时，多达1%的储存RBC经历裂解，并且多达25%的剩余RBC是不可修复的受损的非存活细胞。此外，RBC储存介质积累已知的毒性介质作为RBC代谢和降解的副产物。在输血过程中将这些毒性介质和不可修复的受损细胞输注到接受者体内会降低输血的治疗效果，并导致1-2%的美国公民出现多种不良后果。该项目的目标是设计一种高通量技术，用于在输血过程中实时从储存的RBC单元中在线去除储存介质中不可修复的受损细胞和毒性介质。该项目最初将专注于所提出的技术的设计和性能优化，用于存储红细胞的毫升大小的样本。为了支持设计迭代，我们将开发一种辅助血液学芯片设备，用于以高通量测量数千个单个RBC的所有相关几何、机械和生化特性。在项目的第二阶段，我们将扩大处理临床尺寸RBC单元的优化设计。我们将进行一系列广泛的体外试验，以表征加工RBC的质量。为了进行主要RBC功能的综合测试，我们将开发另一种辅助装置，用于直接测量储存的RBC在人工毛细血管网络中加载/卸载氧气的能力。本项目的预期成果是拟定器械的全尺寸原型，可用于进一步检测经处理、保存良好的储存RBC的促炎和促血栓形成活性，以及人体受试者体内的输血后活力和血管内存活率。常规范例假定袋中所有储存的RBC在储存引起的其性质劣化方面是均匀的，因此试图通过操纵总体储存条件和重新配制添加剂溶液来“复原”储存的RBC。我们的方法通过利用储存的RBC的异质性来挑战这种传统模式，以使仅输注保存良好的细胞，而不受储存介质中不可修复的受损细胞和毒素的影响。这是一种全新的方法，对整个医学实践中输血的安全性和有效性具有潜在的改变游戏规则的变革性影响。 公共卫生相关性：红细胞（RBC）输注是一种挽救生命的疗法，经常在整个医疗实践中对住院患者进行管理。大量输注储存的RBC和输注储存超过2周的RBC单位与并发症增加有关，包括肺衰竭，炎症，血栓，感染和心脏病发作。这个高度创新的项目计划开发一种廉价的、一次性的、高通量的技术，用于在输血过程中从RBC单元中去除这些并发症的已知介质。

DESCRIPTION (provided by applicant): With nearly 15 million units of red blood cells (RBCs) transfused to about 5 million patients in the U.S. every year, RBC transfusion is one of the most commonly prescribed therapies for hospital inpatients. In recent years, ample clinical evidence has accumulated that a significant proportion of morbidity and mortality in critically ill patientsis due to the toxic effects of RBC transfusions. Most transfusions involve RBCs that had been stored in an anticoagulant-preservative solution at 1-6 C for up to 6 weeks. The biochemical, mechanical and functional properties of RBCs deteriorate progressively - by the end of allowable storage up to 1% of stored RBCs undergo lysis, and as many as 25% of the remaining RBCs are irreparably damaged, non-viable cells. Moreover, the RBC storage medium accumulates known mediators of toxicity as byproducts of RBC metabolism and degradation. Infusion of these toxic mediators and the irreparably damaged cells into the recipient during transfusion reduces the therapeutic efficacy of transfusion and contributes to multiple adverse outcomes in 1-2% of U.S. citizens. The goal of this project is to devise a high-throughput technology for in-line removal of irreparably damaged cells and toxic mediators in the storage medium from units of stored RBCs in real-time, during the transfusion process. This project will initially focus on the design and performance optimization of the proposed technology on milliliter-size samples of stored RBCs. To support the design iterations, we will develop an auxiliary hematology-on-a-chip device for measuring all relevant geometric, mechanical and biochemical properties for thousands of individual RBCs at high throughput. At the second stage of the project, we will scale up the optimized design for processing clinical-size RBC units. We will perform a broad panel of in vitro tests to characterize the quality of processed RBCs. To perform an integrative test of the primary RBC function, we will develop another auxiliary device for measuring the ability of stored RBCs to load / offload oxygen in artificial capillary networks directly. The anticipated outcome of this project is a full-scale prototype of the proposed device that can be used to further test the processed well- preserved stored RBCs for pro-inflammatory and pro-thrombotic activity, and post-transfusion viability and intravascular survival in human subjects in vivo. Conventional paradigm postulates that all stored RBCs in a bag are homogeneous with respect to the storage-induced deterioration of their properties, and consequently attempts to 'rejuvenate' stored RBCs through manipulation of the overall storage conditions and re-formulation of additive solutions. Our approach challenges this conventional paradigm by leveraging the heterogeneity of stored RBCs to enable transfusion of only well-preserved cells, free from irreparably damaged cells and toxins in the storage medium. This is an entirely novel approach with a potentially game-changing, transformative impact on the safety and efficacy of transfusions administered throughout the practice of medicine. PUBLIC HEALTH RELEVANCE: Red blood cell (RBC) transfusion is a life-saving therapy frequently administered to hospitalized patients throughout the practice of medicine. Massive transfusions of stored RBCs and transfusions of RBC units stored longer than 2 weeks has been associated with increased complications, including lung failure, inflammation, blood clots, infection and heart attack. This highly-innovative project plans to develop an inexpensive, disposable, high-throughput technology for removal of known mediators of these complications from RBC units during transfusion.