Metabolic markers for mitochondrial function

线粒体功能的代谢标志物

基本信息

批准号：
8218086
负责人：
Cynthia Therese McMurray
金额：
$ 44.02万
依托单位：
UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-20 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8218086
关键词：
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine 1-Methyl-4-phenylpyridinium Abbreviations Address Adenosine Triphosphate Affect Age Amino Acids Animal Model Animals Biological Markers Biology Blood Blood - brain barrier anatomy Brain Calcium Cells Chronic Chronic Disease Clinical Markers Complex Cytosol Defect Detection Disease Early Diagnosis Environment Environmental Exposure Exposure to Functional disorder Heterogeneity Human Individual Injury Intervention Lead Link Mass Spectrum Analysis Measurable Measurement Measures Metabolic Metabolic Diseases Metabolic Marker Methodology Mitochondria Modeling Monoamine Oxidase Nanostructures Noise Oxidation-Reduction Oxidative Phosphorylation Pattern Preventive Intervention Production Property Propionic Acids Research Personnel Resolution Respiration Rodent Rotenone Sampling Signal Transduction Sorting - Cell Movement Staging Symptoms Synaptosomes System Techniques Technology Testing Tissues Toxic Environmental Substances Toxic effect Toxicant exposure Toxin Ubiquitin Urine Variant base brain cell cell motility cell type dopamine transporter functional decline in vivo mitochondrial dysfunction mouse model multicatalytic endopeptidase complex nervous system disorder new technology oxidative damage prevent response tool

项目摘要

DESCRIPTION (provided by applicant): Mitochondrial (MT) dysfunction is a factor in numerous chronic diseases and the toxicity related to environmental exposures, but early deficits in MT function are difficult to detect. Current clinical markers for mitochondrial dysfunction typically detect only advanced symptoms of tissue injury and disease, yet the sensitivity to detect mild MT dysfunction and heterogeneity within tissue has hampered robust identification of meaningful biomarkers at early stages. Mitochondrial biology is variable; and chronic, low level MT dysfunction may be below the detection sensitivity of many techniques. As a result there is the need of new tools to enhance the mechanistic understanding of environmentally-induced mitochondrial toxicity at early stages to enable prevention and intervention. To address this problem, this team of investigators has developed and applied a new technology for single cell mass spectrometry, called Nanostructure-Initiator Mass Spectrometry (NIMS). NIMS has both the single cell resolution (1-10 5m) and the high sensitivity (attomolar) needed to detect early biomarkers of MT dysfunction as metabolic "signatures" in individual cells. NIMS offers a number of advantages over standard mass spectrometry, including (1) ultra-high sensitivity, (2) high selectivity, and (3) single cell resolution to reduce sample complexity. NIMS will be applied to identify metabolic signatures for early MT dysfunction in the brain and blood of diseased animals or animals treated with environmental toxins at "subclinical" levels. In Specific Aim 1, NIMS will be employed to generate metabolic signatures for MT decline. In Specific Aim 2, an activity test will be used to see whether the biomarker reflects functional changes in MT or MT within the context of the cell. NIMS can be applied to any tissue and any cell type, to quantitatively sort out complex changes that occur in dynamic cellular environments, and minimizes the inherent system heterogeneity that has confounded efforts in detecting meaningful markers of MT decline. PUBLIC HEALTH RELEVANCE: Mitochondrial dysfunction is a factor in numerous chronic diseases and environmental exposures, but early deficits in MT function are difficult to detect. To address this problem, we have developed and applied a new technology for single cell mass spectrometry, called NIMS. NIMS has both single cell resolution and the high sensitivity needed to detect early biomarkers of MT dysfunction, and will be used to define "signatures" of early MT dysfunction in single cells. Thus, NIMS technology reduces system heterogeneity, has the high sensitivity needed for detecting meaningful metabolic markers of early MT decline, and the ability to spatially assign signatures in distinct cell types. We apply NIMS identify the effects of oxidative damage in the brain, blood and urine in animal models.

描述（由申请人提供）：线粒体（MT）功能障碍是许多慢性疾病和与环境暴露相关的毒性的一个因素，但MT功能的早期缺陷很难检测到。目前线粒体功能障碍的临床标志物通常只能检测组织损伤和疾病的晚期症状，但检测组织内轻度 MT 功能障碍和异质性的敏感性阻碍了早期阶段对有意义的生物标志物的可靠识别。线粒体生物学是可变的；慢性、低水平 MT 功能障碍可能低于许多技术的检测灵敏度。因此，需要新的工具来在早期阶段加强对环境引起的线粒体毒性的机制理解，以实现预防和干预。为了解决这个问题，该研究小组开发并应用了一种单细胞质谱新技术，称为纳米结构引发剂质谱（NIMS）。 NIMS 具有单细胞分辨率（1-10 5m）和高灵敏度（阿摩尔级），可检测 MT 功能障碍的早期生物标志物作为单个细胞的代谢“特征”。与标准质谱分析相比，NIMS 具有许多优势，包括 (1) 超高灵敏度、(2) 高选择性和 (3) 可降低样品复杂性的单细胞分辨率。 NIMS 将用于识别患病动物或接受“亚临床”水平环境毒素治疗的动物大脑和血液中早期 MT 功能障碍的代谢特征。在具体目标 1 中，NIMS 将用于生成 MT 下降的代谢特征。在具体目标 2 中，将使用活性测试来查看生物标志物是否反映了 MT 或细胞背景下 MT 的功能变化。 NIMS 可应用于任何组织和任何细胞类型，定量分析动态细胞环境中发生的复杂变化，并最大限度地减少固有的系统异质性，这种异质性使检测 MT 下降的有意义的标记物的工作变得混乱。公共卫生相关性：线粒体功能障碍是许多慢性疾病和环境暴露的一个因素，但 MT 功能的早期缺陷很难检测到。为了解决这个问题，我们开发并应用了一种单细胞质谱新技术，称为 NIMS。 NIMS 具有检测 MT 功能障碍早期生物标志物所需的单细胞分辨率和高灵敏度，并将用于定义单细胞中早期 MT 功能障碍的“特征”。因此，NIMS 技术降低了系统异质性，具有检测早期 MT 下降的有意义的代谢标记物所需的高灵敏度，并且能够在不同细胞类型中空间分配特征。我们应用 NIMS 识别动物模型中大脑、血液和尿液氧化损伤的影响。