Proteomics and biomarker

这个项目的重点是假设钙蛋白酶抑制代表了一个合理的和可行的 TBI后治疗干预的目标。创伤性脑损伤（TBI）导致超过50，000例 每年有超过500万美国人死于某种类型的TBI。此外 需要改进的治疗方法，重要的是要了解病理基础的机制， TBI的后遗症一个区域被认为是细胞损伤的重要贡献者， TBI是钙和氧化还原依赖性钙蛋白酶/钙蛋白酶抑制素蛋白水解系统。的中心假设 其核心是钙蛋白酶对特定蛋白质的切割与细胞和神经元的功能障碍有关， 创伤性脑损伤相关的假设是，这种蛋白水解的减弱将导致变性减少， 改善功能结果。鉴于钙蛋白酶在神经变性中的作用的强有力的证据， 长期目标是确定导致TBI后神经变性的钙蛋白酶的靶点。的 本核心的目标是：1）提供TBI后钙蛋白酶底物的统一评估，包括细胞类型 特异性底物，从而更好地表征钙蛋白酶活性的时程和定位; 2） 鉴定和验证神经变性生物标志物以监测损伤进展以及 治疗剂，和; 3）确定氧化应激随时间抑制钙蛋白酶活性的程度 TBI的过程，一个尚未探索的领域。与这三个项目合作，这些核心研究 将利用二维凝胶电泳，质谱，蛋白质印迹分析，和钙蛋白酶活性测定， 脑组织、CSF和血清来实现这些目标。总之，这一核心将提供所需的工具， 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。

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.