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，心血管和衰老有关的关键作用 疾病。但是，线粒体呼吸测定法当前需要处理活组织 从患者带走后一个小时内采样。该要求使呼吸测定法分析很大程度上 对标准临床实践和临床研究不可行。我们发明了一项新技术来评估最大 以前冷冻的生物样品中的线粒体呼吸能力，到目前为止被认为 科学界不可能。并排比较证实了我们的测定准确反映 结果在新鲜样品中测量。因此，我们的技术克服了标准的基本限制 接近需要生命组织的方法。此外，我们的方法包括简化的样本准备， 使用的生物材料少得多。 Enspire Bio将进一步开发这种专利的冷冻丝图技术以实现呼吸测定法 目前尚不可用的临床应用和冷冻样品的测量。我们将实现 我们的目标1：建立从冷冻组织中的氧气消耗测量的标准方案 源自人和小鼠组织样品的样品。我们将通过启用 从组织匀浆而不是分离的线粒体中消耗氧的测量。目标2： 建立专有套件并将其交付到五个Beta测试网站，包括舆论学术界 实验室，线粒体疾病中心以及一家生物技术和制药公司，以测试和提供反馈。 为了向没有湿实验室测定能力的客户提供冷冻样本分析，我们将开发一个Cro- 在内部键入服务，其中将运送样品来启动Pirebio进行分析。与一组飞行员一起工作 学术，临床和行业客户将提供反馈以建立程序和经济学 冷冻组织呼吸测定法的CRO模型方法。