Proteomics and biomarker

蛋白质组学和生物标志物

• 批准号: 8058717
• 负责人: RODNEY P GUTTMANN
• 金额: $ 20.02万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8058717
• 关键词: 4 hydroxynonenal; Address; Alzheimer' s Disease; American; Amyotrophic Lateral Sclerosis; Animal Model; Area; Astrocytes; Behavioral; Biological Assay; Biological Markers; Biological Models; Brain Injuries; Calcium; Calpain; Cell Culture Techniques; Cell Death; Cells; Cerebral Ischemia; Cerebrospinal Fluid; Cessation of life; Cleaved cell; Collaborations; Data; Digestion; Disease; Effectiveness; Enzymes; Functional disorder; Future; Goals; Huntington Disease; In Vitro; Individual; Injury; Ionophores; Knockout Mice; Link; Lipid Peroxidation; Mass Spectrum Analysis; Measurement; Measures; Mediating; Methods; Microglia; Mitochondria; Modeling; Monitor; Nature; Nerve Degeneration; Neuroglia; Neuronal Dysfunction; Neurons; Oligodendroglia; Outcome; Oxidation-Reduction; Oxidative Stress; Parkinson Disease; Pathway interactions; Peptide Hydrolases; Population; Primary Cell Cultures; Prions; Protein Isoforms; Proteins; Proteolysis; Proteomics; Publishing; Role; Serum; Spectrin; Spinal cord injury; System; Therapeutic; Therapeutic Intervention; Time; Traumatic Brain Injury; Treatment Efficacy; Two-Dimensional Gel Electrophoresis; Western Blotting; Work; attenuation; base; brain tissue; calpastatin; cell injury; cell type; disability; efficacy testing; functional outcomes; improved; in vivo; inhibitor/antagonist; insight; m-calpain; neurofilament; novel; oxidation; prevent; programs; small molecule; therapeutic effectiveness; tool; working group

This Program Project focuses on the hypothesis that calpain inhibition represents a rationale and feasible target for therapeutic intervention following TBI. Traumatic brain injury (TBI) results in more than 50,000 deaths each year with over 5 million Americans suffering the effects of some type of TBI. In addition the need for improved therapeutics, it is important to understand the mechanisms underlying the pathological sequelae to TBI. One area that has been implicated as an important contributor to cellular injury following TBI is the calcium- and redox-dependent calpain/calpastatin proteolytic system. The central hypothesis of this core is that calpain cleavage of specific proteins is linked to cell and neuronal and dysfunction following TBI. The related hypothesis is that attenuation of this proteolysis will result in reduced degeneration and improved functional outcome. Given the strong evidence for the role of calpains in neurodegeneration, the long-term goal is to identify the targets of calpain that contribute to neurodegeneration following TBI. The goals of this Core are to: 1) provide uniform assessment of calpain substrates following TBI, including celltype specific substrates, thereby better characterizing the time-course and localization of calpain activity; 2) identify and verify biomarkers of neurodegeneration to monitor injury progression as well as efficacy of therapeutics, and; 3) determine the extent to which oxidative stress inhibits calpain activity over the time course of TBI, an area that remains unexplored. In collaboration with the three projects, these core studies will utilize 2D-gel electrophoresis, mass-spectrometry, Western blot analysis, and calpain activity assays with brain tissue, CSF, and serum to achieve these goals. In summary, this core will provide the needed tools and assistance for uniform analysis of calpain substrate degradation within the projects, provide additional support for the discovery of calpain substrates and related biomarkers, and provide direct measurements of calpain activity to determine the influence of oxidative stress on calpain activity following TBI.

该计划项目的重点是钙蛋白酶抑制这一假设是合理且可行的 TBI 后治疗干预的目标。创伤性脑损伤 (TBI) 导致 50,000 多人死亡 每年有超过 500 万美国人遭受某种类型的 TBI 影响而死亡。此外 需要改进的治疗方法，了解病理基础的机制很重要 TBI 的后遗症。一个被认为是细胞损伤的重要贡献者的区域 TBI 是钙和氧化还原依赖性钙蛋白酶/钙蛋白酶抑制剂蛋白水解系统。中心假设为 这个核心是钙蛋白酶对特定蛋白质的裂解与细胞和神经元以及随后的功能障碍有关 创伤性脑损伤。相关的假设是，这种蛋白水解作用的减弱将导致变性减少和 改善功能结果。鉴于钙蛋白酶在神经退行性疾病中的作用有强有力的证据， 长期目标是确定钙蛋白酶导致 TBI 后神经退行性变的靶标。这 该核心的目标是：1) 在 TBI 后提供钙蛋白酶底物的统一评估，包括细胞类型 特定底物，从而更好地表征钙蛋白酶活性的时间过程和定位； 2） 识别和验证神经退行性变的生物标志物，以监测损伤进展以及疗效 治疗学，以及； 3) 确定一段时间内氧化应激抑制钙蛋白酶活性的程度 TBI 的过程，这是一个尚未探索的领域。这些核心研究与这三个项目合作 将利用 2D 凝胶电泳、质谱、蛋白质印迹分析和钙蛋白酶活性测定 脑组织、脑脊液和血清来实现这些目标。总之，该核心将提供所需的工具和 协助项目内钙蛋白酶底物降解的统一分析，提供额外的 支持钙蛋白酶底物和相关生物标志物的发现，并提供直接测量 钙蛋白酶活性以确定 TBI 后氧化应激对钙蛋白酶活性的影响。