Metabolic markers for mitochondrial function

线粒体功能的代谢标志物

基本信息

批准号：
8895766
负责人：
Cynthia Therese McMurray
金额：
$ 51.36万
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 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8895766
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）功能障碍是多种慢性疾病的一个因素，毒性与环境暴露，但很难检测到MT功能的早期定义。当前的临床标记线粒体功能障碍通常仅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强大的鉴定在早期阶段有意义的生物标志物。线粒体生物学是可变的。和慢性，低水平MT功能障碍可能低于许多技术的检测灵敏度。我们需要新工具来增强机械了解在早期阶段对环境引起的线粒体毒性的理解，以实现预防和干涉。为了解决这个问题，我们已经开发并应用了一项新技术作为单细胞质量光谱法，称为纳米结构 - 引导剂质谱法（NIMS）。 NIMS具有单个单元分辨率（1-10€）和需要检测MT功能障碍的早期生物标志物所需的高灵敏度（Attomolor）作为单个细胞中的代谢“特征”。 NIMS比标准质量提供了许多优势光谱法，包括（1）超高灵敏度，（2）高选择性和（3）单细胞分辨率以降低样品复杂性。我们应用NIMS来识别大脑早期MT功能障碍的代谢特征，在“亚临床”水平上用环境毒素治疗的患病动物或动物的血液。在AIM 1中，我们使用 NIMS产生代谢特征以减少MT。在AIM 2中，我们将在功能上测试生物标志物是否反映在单元格中MT或MT的功能变化。 NIM可以应用于任何组织，任何细胞类型，以定量地分解在动态细胞环境中发生的复杂变化，以及最小化继承系统异质性，这使人们对检测有意义的标记进行了困惑 MT下降。