High subzero heart preservation: from zebrafish to mammals

高度零下心脏保存：从斑马鱼到哺乳动物

• 批准号: 10601049
• 负责人: Shannon Noella Tessier
• 金额: $ 42万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10601049
• 关键词: 3-Dimensional; ANXA5 gene; Address; Adult; Animal Experimentation; Animal Model; Animals; Apoptosis; Architecture; Biological Assay; Calcium; Cardiac; Cardiac Myocytes; Cell Survival; Cell model; Cells; Cessation of life; Chemicals; Clinical; Complex; Dependence; Development; Emergency Situation; Endothelial Cells; Exposure to; Freezing; Generations; Heart; Heart Diseases; Heart Research; Heart Transplantation; Heat-Shock Proteins 70; Homeostasis; Hour; Hypoxia; Hypoxia Inducible Factor; Ice; In Vitro; Injury; Ischemia; Larva; Length; Libraries; Life; Life Expectancy; Literature; Location; Mammals; Metabolic; Methods; Modeling; Molecular; Myocardial Ischemia; Myofibrils; Nature; Operative Surgical Procedures; Organ; Organ Donor; Outcome; Outcome Assessment; Patient-Focused Outcomes; Patients; Perfusion; Procedures; Protective Agents; Proteins; Protocols documentation; Rana; Rattus; Reporter; Research; Resources; Risk; Signal Transduction; Solid; Structure; Suspensions; System; Technology; Temperature; Time; Tissues; Transgenic Organisms; Translations; Transplantation; United States; Waiting Lists; Wood material; Work; Zebrafish; cell type; clinically relevant; cost; cryogenics; design; experimental study; functional outcomes; heart function; heart preservation; improved; ischemic injury; model organism; mutant; natural hypothermia; novel; preconditioning; preservation; response; scale up; stressor; success; surgery outcome; tool; transcription factor; usability

PROJECT SUMMARY Heart transplantation is the only cure for end-stage heart disease; however, an increasing imbalance between demand and supply has limited access to this life-saving procedure. Extending the length of time hearts can remain alive during transport represents a critical enabling technology to address several limitations in heart transplantation. For example, usable hearts are discarded due to circumstantial factors, such as the timing of donor death and donor/recipient location, which can be eliminated by extended preservation. Further, longer preservation duration has the potential to improve recipient outcomes by enhancing donor-recipient matching, reducing ischemic injury, and removing the risk of emergency surgeries. Consequently, this project aims to develop a new method to extend preservation of hearts for transplantation from 4-6 hours to 3+ days. While we have mastered the preservation of several clinically relevant cell types in suspension, solid organs have been met with limited success since different cell types have different responses to the same injury. Further, high throughput experimental methods to aid development of preservation solutions are limited since intact tissue has 3D structural considerations that have a profound effect on successful preservation and commonly used in vitro cell models are not representative and lack assessment of functional outcomes. Instead, we propose to strategically leverage zebrafish transgenic lines to address cell-specific responses, functional consequences, and compounding injuries of preservation approaches with an intact, native heart in a high throughout format. Approaches developed in zebrafish will be rapidly scaled to mammalian hearts to promote translation. We will leverage this experimental platform to enable the development of a new preservation approach, termed partial freezing. Inspired by freeze-tolerant wood frogs in nature, partial freezing aims to maximize storage durations by entering unexplored high subzero temperatures ranges approaching -20°C. To enter these temperature ranges, we will develop effective strategies for hearts to live in the presence of ice. This will be achieved through the development of a storage solution designed to protect diverse cardiac cell types without adverse functional consequences (Specific Aim 1). These efforts will be complimented by the discovery of inducers that activate hypoxia signaling (Specific Aim 2) and the unfolded protein response (Specific Aim 3) to rescue hearts from ischemic and temperature-dependent injury, respectively. These inducers will be combined into a preconditioning solution that is delivered during machine perfusion to further extend preservation time.

项目摘要 心脏移植是治疗终末期心脏病的唯一方法;然而， 供求关系限制了获得这一救生程序的机会。延长心脏的时间长度 在运输过程中保持存活是一项关键的使能技术，可以解决心脏的几个限制。 移植例如，可用的心脏由于环境因素而被丢弃，例如心脏移植的时机。 供体死亡和供体/受体的位置，这可以通过延长保存来消除。更远，更长 保存时间具有通过增强供体-受体匹配来改善受体结果的潜力， 减少缺血性损伤，消除紧急手术的风险。因此，该项目旨在 开发一种新的方法，将移植用心脏的保存时间从4-6小时延长到3天以上。 虽然我们已经掌握了在悬浮液中保存几种临床相关细胞类型，但实体器官 由于不同的细胞类型对相同的损伤有不同的反应，因此成功有限。 此外，帮助开发保存溶液的高通量实验方法受到限制，因为 完整组织具有3D结构考虑，其对成功保存具有深远影响， 通常使用的体外细胞模型不具有代表性，并且缺乏对功能结果的评估。 相反，我们建议战略性地利用斑马鱼转基因系来解决细胞特异性反应， 功能性后果，以及保存方法与完整的天然心脏的复合损伤， 一种高通量的格式。在斑马鱼中开发的方法将迅速扩展到哺乳动物心脏， 促进翻译。 我们将利用这个实验平台，开发一种新的保存方法，称为 部分冷冻。受自然界中耐寒树蛙的启发，部分冷冻旨在最大限度地储存 持续时间通过进入未经探索的高零下温度范围接近-20 ° C。进入这些 温度范围，我们将制定有效的策略，使心脏在冰的存在下生存。这将是 通过开发一种储存解决方案来实现，该解决方案旨在保护各种心脏细胞类型， 不良功能后果（具体目标1）。这些努力将得到以下发现的补充： 激活缺氧信号传导（特异性目标2）和未折叠蛋白反应（特异性目标3）的诱导剂， 分别从缺血性和温度依赖性损伤中拯救心脏。这些诱导剂将结合在一起 在机器灌注期间输送预处理溶液以进一步延长保存时间。