Metabolic markers for mitochondrial function

线粒体功能的代谢标志物

基本信息

批准号：
8697051
负责人：
Cynthia Therese McMurray
金额：
$ 50.87万
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/8697051
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 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

ABSTRACT 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. We need new tools to enhance the mechanistic understanding of environmentally-induced mitochondrial toxicity at early stages to enable prevention and intervention. To address this problem, we have 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 ¿m) 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. We apply NIMS 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 Aim 1, we use NIMS to generate metabolic signatures for MT decline. In Aim 2, we will functionally test 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.

抽象的线粒体 (MT) 功能障碍是许多慢性疾病及其相关毒性的一个因素环境暴露，但 MT 功能的早期缺陷很难检测到。线粒体功能障碍通常只能检测到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对早期阶段有意义的生物标志物是可变的；以及慢性、低水平的 MT 功能障碍。可能低于许多技术的检测灵敏度，我们需要新的工具来增强机械性能。在早期阶段了解环境引起的线粒体毒性，以便能够预防和为了解决这个问题，我们开发并应用了一种单细胞群新技术。光谱测定法，称为纳米结构引发剂质谱法（NIMS），具有单细胞。检测 MT 功能障碍的早期生物标志物所需的分辨率（1-10 µm）和高灵敏度（阿摩尔）作为单个细胞的代谢“特征”，NIMS 比标准质量具有许多优势。光谱分析，包括 (1) 超高灵敏度，(2) 高选择性，以及 (3) 单细胞分辨率，以降低我们应用 NIMS 来识别大脑中早期 MT 功能障碍的代谢特征。在目标 1 中，我们使用患病动物或接受“亚临床”水平环境毒素处理的动物的血液。 NIMS 生成 MT 下降的代谢特征在目标 2 中，我们将功能测试生物标志物是否存在。 MT 或 MT 在细胞范围内的功能变化可以应用于任何组织和细胞。任何细胞类型，定量分析动态细胞环境中发生的复杂变化，以及最大限度地减少固有的系统异质性，这种异质性混淆了检测有意义的标记物的努力 MT下降。