Proteomics and biomarker

该计划项目侧重于这样一种假设，即抑制Calain代表了一种合理和可行的 颅脑损伤后治疗干预的靶点。创伤性脑损伤(TBI)导致超过50,000人 每年有500多万美国人死于某种类型的脑外伤。此外， 需要改进的治疗方法，重要的是要了解潜在的病理机制 脑损伤的后遗症。一个被认为是导致以下细胞损伤的重要因素的区域 TBI是依赖于钙离子和氧化还原的钙蛋白酶/钙调蛋白蛋白分解系统。的中心假说 这一核心是特定蛋白的钙蛋白酶的切割与细胞和神经元以及随后的功能障碍有关。 TBI。相关的假设是，这种蛋白质分解的减弱将导致减少退化和 改善了功能结果。鉴于有强有力的证据表明钙调蛋白在神经退行性变中的作用， 长期目标是确定导致脑外伤后神经退行性变的钙蛋白酶的靶点。这个 这个核心的目标是：1)提供对脑损伤后钙蛋白底物的统一评估，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 治疗学；以及；3)确定氧化应激在一定时间内抑制钙蛋白酶活性的程度 TBI的进程，这是一个仍未勘探的领域。与这三个项目合作，这些核心研究 将利用2D-Gel电泳法、质谱仪、Western印迹分析和Calain活性分析 脑组织、脑脊液和血清来实现这些目标。总而言之，这个核心将提供所需的工具和 协助统一分析项目内钙蛋白酶底物的降解情况，提供额外的 支持发现钙蛋白底物和相关生物标记物，并提供直接测量 测定氧化应激对颅脑损伤后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.