A quantitative viability metric for liver transplantation using Resonance Raman Spectroscopy

使用共振拉曼光谱进行肝移植的定量活力指标

• 批准号: 10562740
• 负责人: Shannon Noella Tessier
• 金额: $ 51.46万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10562740
• 关键词: Acute; Allografting; Animals; Biopsy; Cardiac Death; Cells; Cessation of life; Chronic; Clinical; Compensation; Data; Devices; Disease; Exposure to; Family suidae; Functional disorder; Goals; Heart; Human; Improve Access; Industry Collaboration; Injury; Intervention; Life; Liver; Logistic Regressions; Mammals; Measurement; Measures; Methodology; Methods; Mitochondria; Modeling; Operative Surgical Procedures; Organ; Organ Donations; Organ Donor; Organ Transplantation; Organ Viability; Organ failure; Outcome; Oxidation-Reduction; Patients; Perfusion; Play; Raman Spectrum Analysis; Rattus; Recovery; Reperfusion Injury; Research; Research Personnel; Resources; Risk; Rodent; Rodent Model; Role; Sampling; Scientist; Surface; Surgeon; Survival Rate; Technology; Time; Tissues; Translating; Transplantation; Validation; Waiting Lists; Warm Ischemia; Work; arm; attributable mortality; body system; conditioning; cost; end-stage organ failure; flexibility; imaging platform; indexing; injured; innovation; liver biopsy; liver injury; liver transplantation; meetings; novel; organ allocation; organ injury; post-transplant; preclinical study; response; success; tissue injury

PROJECT SUMMARY Organ transplantation is a life-saving treatment for end-stage organ disease; however, there is a severe shortage of donor organs whereby only ~36% of wait-listed patients receive a transplant. At the same time, many vital organs that may be transplantable are discarded - some estimates suggest that up to 25% of recovered organs are not ultimately transplanted, and there are additional pools of unrecovered organs that are only marginally damaged. However, because of their uncertain viability and the high cost of an unsuccessful transplant, none of these potential organs are used while transplantation of just a fraction could dramatically reduce the organ shortage. A particularly large target of untapped donor organs are marginally injured warm ischemic organs from Donation after Circulatory Death (DCD). While these organs have been correlated to lower survival rates and increased post-transplant complications, machine perfusion technology holds the promise of reconditioning some of these organs, thereby dramatically increasing availability. However, a critical bottleneck that impacts experimental and clinical advances in reconditioning organs are developing non-objective metrics that can definitively ascertain if an organ is viable prior to transplantation. Clinically, since organ allocation is already severely restricted by time, these criteria need to be defined for each organ rapidly and preferably in real-time. Experimentally, access to flexible platforms would increase the scale and number of researchers that can tackle these difficult problems. In response to this need, we propose to develop a novel imaging platform that will use Resonance Raman Spectroscopy (RRS) to quantify mitochondrial redox state on tissue surfaces/biopsies and mitochondrial breakdown products in the perfusate. Our approach is unique in organ transplantation assessment since: 1) results are obtained in less than 3 minutes and are non-destructive, and 2) offers the flexibility required for diverse organ systems, significantly increasing the impact of the proposed work. While we will target diverse transplantable organs, we focus our initial efforts on liver through execution of two specific aims. We will develop a benchtop RRS device with the capacity to integrate several types of measurements on livers, including biopsies, surface measurements, and perfusate samples. In Specific Aim 1, we will apply and validate the biopsy cell and extendable arm that enables liver biopsies and surface measurements to calculate the ratio of reduced to total mitochondria (defined as Resonance Raman Reduced Mitochondrial Ratio, 3RMR). In Specific Aim 2, we will apply and validate the perfusate cell that is compatible with the same benchtop RRS device to quantify mitochondrial breakdown products in real-time from the perfusate (defined as the Perfusate Viability Index, PVI). In SA1-2, we will define the 3RMR/PVI threshold values that if exceeded will indicate irreversible liver injury, beginning with rigorous studies in rodents before measuring samples from discarded human DCD livers during machine perfusion.

项目概要 器官移植是终末期器官疾病的挽救生命的治疗方法；但严重短缺 的捐赠器官，只有约 36% 的等待名单患者接受移植。与此同时，许多重要的 可移植的器官被丢弃——一些估计表明，高达 25% 的回收器官被丢弃 未最终移植，并且还有其他未恢复的器官池，这些器官仅被少量移植 损坏的。然而，由于它们的生存能力不确定，而且移植不成功的成本很高，没有一种 这些潜在的器官被利用，而仅移植一小部分就可能大大减少器官的数量 短缺。 未开发的供体器官的一个特别大的目标是受到轻微损伤的热缺血器官。 循环死亡后捐赠（DCD）。虽然这些器官与较低的存活率和 移植后并发症增加，机器灌注技术有望实现修复 其中一些器官，从而大大提高了可用性。然而，一个影响的关键瓶颈 器官修复的实验和临床进展正在开发非客观指标，可以 在移植前明确确定器官是否可行。临床上由于器官分配已经 由于时间的严格限制，需要为每个器官快速定义这些标准，最好是实时定义。 从实验上看，使用灵活的平台将增加能够解决问题的研究人员的规模和数量 这些难题。为了满足这一需求，我们建议开发一种新颖的成像平台，该平台将使用 共振拉曼光谱 (RRS) 可量化组织表面/活检中的线粒体氧化还原状态 灌注液中的线粒体分解产物。我们的方法在器官移植评估方面是独一无二的 因为：1) 不到 3 分钟即可获得结果，并且是非破坏性的，2) 提供所需的灵活性 对于不同的器官系统，显着增加拟议工作的影响。 虽然我们的目标是多种可移植器官，但我们最初的努力集中在肝脏上，通过执行两项 具体目标。我们将开发一种台式 RRS 设备，能够集成多种类型的 肝脏测量，包括活检、表面测量和灌注液样本。在具体目标 1 中，我们 将应用并验证活检室和可伸展臂，以实现肝脏活检和表面测量 计算还原线粒体与总线粒体的比率（定义为共振拉曼还原线粒体 比率，3RMR）。在具体目标 2 中，我们将应用并验证与相同的灌流液细胞兼容的灌流液细胞。 台式 RRS 装置，用于实时定量灌注液中的线粒体分解产物（定义为 灌注液活力指数 (PVI)。在 SA1-2 中，我们将定义 3RMR/PVI 阈值，如果超过 将表明不可逆的肝损伤，首先对啮齿类动物进行严格的研究，然后再测量来自 在机器灌注过程中丢弃人类 DCD 肝脏。