MITOCHONDRIAL RESPIROMETRY IN FROZEN BIOLOGICAL SAMPLES

冷冻生物样品中的线粒体呼吸测定

• 批准号: 10011475
• 负责人: Orian S Shirihai
• 金额: $ 22.5万
• 依托单位: ENSPIRE BIO, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-20 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10011475
• 关键词: Academia; Acute; Aging; Biocompatible Materials; Biological; Biological Assay; Biotechnology; Cardiovascular system; Cells; Client; Clinical; Clinical Research; Communities; Complex; Data; Development; Disease; Drug Exposure; Economics; Environment; Environmental Monitoring; Evaluation; Feedback; Freezing; Future; Genetic Models; Goals; Hour; Human; Impairment; Industry; Instruction; Legal patent; Measurement; Measures; Metabolic; Methodology; Methods; Mitochondria; Mitochondrial Diseases; Monitor; Mus; Organizational Models; Outcome; Oxygen Consumption; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Play; Population; Positioning Attribute; Preparation; Procedures; Protocols documentation; Reagent; Reproducibility; Research; Research Contracts; Research Personnel; Respiration; Respiratory physiology; Running; Sampling; Services; Ships; Side; Site; Swab; Technology; Technology Assessment; Testing; Time; Tissue Sample; Tissues; Toxicant exposure; Toxicology; United States Food and Drug Administration; clinical application; clinical development; clinical practice; clinically relevant; environmental toxicology; improved; minimally invasive; new technology; respiratory; toxicant

Abstract Impaired mitochondrial respiration plays a key role in metabolic, cardiovascular, and aging-related diseases. However, mitochondrial respirometry analysis currently requires processing of the living tissue sample within an hour after being taken from the patient. This requirement makes respirometry analysis largely unfeasible to standard clinical practice and clinical studies. We invented a new technology to assess maximal mitochondrial respiratory capacity in previously-frozen biological samples, which thus far been considered impossible by the scientific community. Side-by-side comparison confirmed that our assay accurately reflects results measured in fresh samples. Our technology thus overcomes the fundamental limitation of standard approaches that required living tissue. Moreover, our approach consists of simplified sample preparation which uses significantly less biological material. Enspire Bio will further develop this patented Frozen Mitos technology to enable respirometry measurements for clinical applications and frozen samples, which currently is not available. We will achieve our goals by Aim 1: Establishing standard protocols for oxygen consumption measurements from frozen tissue samples derived from human and mouse tissue samples. We will further improve throughput by enabling measurements of oxygen consumption from tissue homogenate instead of isolated mitochondria. Aim 2: Establishing proprietary kits and delivering those to five beta testing sites including opinion-leading academic labs, mitochondrial disease centers and a biotech and pharmaceutical company to test and provide feedback. To provide frozen sample analysis to clients that have no capacity for wet lab assays, we will develop a CRO- type service in-house where samples will be shipped to EnspireBio for analysis. Working with a set of pilot academic, clinical and industry clients will provide feedback to establish procedures and the economics of the CRO model approach for the frozen tissue respirometry.

抽象的 线粒体呼吸受损在代谢、心血管和衰老相关方面发挥着关键作用 疾病。然而，线粒体呼吸测定分析目前需要对活体组织进行处理 从患者体内采集样本后一小时内进行。这一要求使得呼吸测量分析在很大程度上 不适合标准临床实践和临床研究。我们发明了一项新技术来评估最大 先前冷冻的生物样本中的线粒体呼吸能力，迄今为止被认为 科学界不可能。并排比较证实我们的测定准确地反映了 在新鲜样品中测量的结果。因此，我们的技术克服了标准的根本限制 需要活组织的方法。此外，我们的方法包括简化的样品制备， 使用的生物材料显着减少。 Enspire Bio 将进一步开发这项获得专利的 Frozen Mitos 技术，以实现呼吸测量 用于临床应用和冷冻样本的测量，目前尚不可用。我们将实现 我们的目标 目标 1：建立冷冻组织耗氧量测量的标准协议 来自人类和小鼠组织样本的样本。我们将通过启用来进一步提高吞吐量 测量组织匀浆而不是分离的线粒体的耗氧量。目标 2： 建立专有套件并将其交付给五个 Beta 测试站点，其中包括意见领先的学术站点 实验室、线粒体疾病中心以及生物技术和制药公司进行测试并提供反馈。 为了向没有湿实验室检测能力的客户提供冷冻样本分析，我们将开发一个 CRO- 内部类型服务，样品将被运送到 EnspireBio 进行分析。与一组试点合作 学术、临床和行业客户将提供反馈，以建立程序和经济性 用于冷冻组织呼吸测量的 CRO 模型方法。