High subzero preservation of liver for transplantation

移植用肝脏的高度低温保存

• 批准号: 9898358
• 负责人: Mehmet Toner
• 金额: $ 38.48万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-17 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9898358
• 关键词: Animals; Bioinformatics; Biomedical Engineering; Biomimetics; Blood Vessels; Blood capillaries; Body Water; Breathing; Cell Survival; Cells; Clinical; Clinical Pathways; Crystallization; Data; Dehydration; Development; Discontinuous Capillary; Encapsulated; Endothelial Cells; Energy Metabolism; Engineering; Ensure; Evaluation; Extracellular Space; Freezing; Glycocalyx; Hepatocyte; Hour; Human; Ice; Infiltration; Infrastructure; Length; Liquid substance; Literature; Liver; Metabolic; Methods; Microfluidics; Modeling; Molecular; Molecular Profiling; Names; Nature; Operative Surgical Procedures; Organ; Organ Donor; Organ Preservation; Organ Transplantation; Outcome; Pathway interactions; Patients; Pharmacology; Phenotype; Play; Process; Property; Protocols documentation; Public Health; Rana; Rattus; Regulatory Pathway; Reperfusion Therapy; Research; Role; Screening procedure; Spatial Distribution; Technology; Temperature; Testing; Time; Tissues; Translating; Transplantation; United States; Waiting Lists; Work; animation; base; body system; cell injury; cost; extracellular; improved; interdisciplinary approach; liver preservation; liver transplantation; novel strategies; organ injury; phase change; preservation; prevent; programs; scale up; stress tolerance; success; tissue/organ preservation; visual tracking; wasting

PROJECT SUMMARY With more than five times the number of patients on the wait list than will receive a donor organ in the United States, the field of transplantation is facing a serious donor shortage crisis. Overcoming the organ shortage will require integrated strategies, including a particular focus on overcoming ineffective bio-preservation and stabilization protocols. Longer storage durations will provide the infrastructure required to enable global matching programs, eliminate the need to scramble and conduct unplanned surgeries, and reduce unnecessary waste of quality organs. We believe the method for preserving mammalian organs should employ hibernating and freeze-tolerant strategies in nature that are then further augmented using bioengineering principles. Consequently, we seek to develop a protocol for human organ preservation which will achieve high subzero storage temperatures (ranging from -10 to -20 °C) in the presence of extracellular ice, and storage durations of weeks to months, using inspiration from in nature. Our approach is unique in organ/tissue preservation literature since we aim to actively initiate ice propagation in the vasculature and extracellular spaces, rather than extreme means of inhibiting ice crystallization as is the current standard. The presence of non-injurious ice will be essential in achieving longer storage durations, while also playing an important role in the scale-up to human livers. While this program targets the banking of human liver, our discoveries and solutions will be translatable to other tissues and organ systems. In Specific Aim 1, we will adapt endothelial cell-coated microvascular networks already developed by our group3 in order to model and develop strategies to overcome challenges associated with ice propagation. Since endothelial cells in the vasculature will be the most vulnerable to ice propagation, SA#1 will be an essential proof of concept of our novel strategy and we already have promising data. In Specific Aim 2, we will engineer an ice nucleating agent which will promote non-injurious propagation of ice in extracellular spaces. Ice nucleating agents are essential for restricting ice formation to extracellular spaces and have been identified as critical strategies for freezing survival. In Specific Aim 3, we will reprogram cells to descend into a state of `suspended animation' with enhanced stress tolerance, as inspired by nature. We will achieve this using both passive temperature effects as well as using pharmacological agents. We will perform in-depth characterization of the molecular impact of our cellular reprogramming efforts. In each specific aim, we scale up rapidly to rat whole liver while also validating in human livers in order to maximize impact.

项目摘要 等待名单上的患者数量是美国接受捐赠器官的患者数量的五倍多 美国，移植领域正面临严重的供体短缺危机。克服器官短缺将 需要综合战略，包括特别注重克服无效的生物保护， 稳定协议。更长的存储持续时间将提供实现全球 匹配的程序，消除了争夺和进行计划外手术的需要，并减少了不必要的 浪费优质器官。 我们认为哺乳动物器官的保存方法应该采用冬眠和抗冻的方法 然后利用生物工程原理进一步增强。因此，我们力求 制定一项人体器官保存协议，以实现零度以下的高储存 温度（范围从-10至-20 °C）在细胞外冰的存在下， 数周到数月，利用大自然的灵感。我们的方法在器官/组织保存方面是独一无二的 文献，因为我们的目标是积极启动冰在血管和细胞外空间的传播，而不是 而不是像目前的标准那样抑制冰结晶的极端手段。非伤害性的存在 冰在实现更长的储存时间方面将是必不可少的，同时在扩大规模方面也发挥着重要作用。 人类的肝脏虽然该计划的目标是人类肝脏的银行，但我们的发现和解决方案将 可转移到其他组织和器官系统。 在具体目标1中，我们将调整我们已经开发的内皮细胞包被的微血管网络， 第三组，以模拟和制定战略，以克服与冰传播有关的挑战。 由于血管系统中的内皮细胞将最易受冰传播的影响，因此SA#1将是一个 这是我们新策略的基本概念证明，我们已经有了有希望的数据。在第二阶段，我们将 设计一种冰成核剂，该冰成核剂将促进冰在细胞外空间中的非伤害性传播。 冰成核剂对于限制冰在细胞外空间的形成至关重要，并且已经被确定 作为冷冻生存的关键策略。在具体目标3中，我们将重新编程细胞，使其下降到 “假死”与增强的压力耐受性，灵感来自大自然。我们将同时使用 被动温度效应以及使用药理学试剂。我们将深入执行 我们的细胞重编程工作的分子影响的表征。在每一个具体目标中， 快速上升到大鼠整个肝脏，同时也在人类肝脏中验证，以最大限度地发挥影响。