High subzero preservation of liver for transplantation

项目总结 等待名单上的患者数量是美国接受捐献器官患者数量的五倍多 在美国，移植领域正面临严重的捐赠者短缺危机。克服器官短缺将会 需要综合战略，包括特别注重克服无效的生物保护和 稳定方案。更长的存储持续时间将提供实现全球 匹配计划，消除争先恐后和进行计划外手术的需要，并减少不必要的 浪费优质器官。 我们认为保存哺乳动物器官的方法应该是冬眠和耐寒。 自然界中的战略，然后利用生物工程原理进一步加强。因此，我们寻求 制定一种人体器官保存协议，实现高低温保存 细胞外有冰时的温度(从-10°C到-20°C)，以及 从几周到几个月，从大自然中获得灵感。我们的方法在器官/组织保存方面是独一无二的 文献，因为我们的目标是在血管和细胞外空间主动启动冰的传播，而不是 而不是像目前的标准那样用极端的手段抑制冰结晶。无伤大雅的存在 冰将是实现更长存储持续时间的关键，同时也在纵向扩展中发挥重要作用 对人类肝脏来说。虽然这个项目的目标是人体肝脏银行，但我们的发现和解决方案将是 可翻译到其他组织和器官系统。 在具体目标1中，我们将使我们已经开发的内皮细胞包裹的微血管网络 第三组，以便建立模型并制定战略，以克服与冰传播有关的挑战。 由于血管系统中的内皮细胞最容易受到冰传播的影响，SA#1将是一种 我们新战略的基本概念证明，我们已经有了令人振奋的数据。在具体目标2中，我们将 设计一种冰核剂，它将促进冰在细胞外空间的非破坏性传播。 冰核剂是将冰形成限制在细胞外空间所必需的，已经被确定 作为冷冻生存的关键策略。在特定目标3中，我们将对单元进行重新编程，使其下降到 在自然的启发下，增强了对压力的耐受性，从而实现了“暂停生命”。我们将使用两种方法来实现这一点 被动的温度效应以及使用药物。我们将深入表演 描述我们的细胞重新编程努力的分子影响。在每个特定的目标中，我们都在 迅速上升到大鼠全肝，同时也在人体肝脏中验证，以最大限度地发挥作用。

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.