Role of calpastatin in modulating function following TBI

钙蛋白酶抑制剂在 TBI 后功能调节中的作用

• 批准号: 7288118
• 负责人: KATHRYN E SAATMAN
• 金额: $ 16.6万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7288118
• 关键词: Acute; Address; Attenuated; Behavioral; Biological Availability; Brain; Brain Injuries; Calcium; Calpain; Cell Count; Cell Death; Cell Death Signaling Process; Cell Survival; Cells; Cerebrospinal Fluid; Cessation of life; Cognitive; Condition; Cytoskeletal Proteins; Data; Diffuse; Diffuse Brain Injury; Disease; Disinhibition; Disruption; Dose; Drops; Evaluation; Event; Functional disorder; Genetic; Glutamate Receptor; Goals; Hippocampus (Brain); Human; Immunoblotting; Immunohistochemistry; Impairment; In Vitro; Injury; Knock-out; Knockout Mice; Maintenance; Mediating; Membrane Proteins; Mitochondria; Modeling; Molecular; Morbidity - disease rate; Motor; Mus; Nerve Degeneration; Neurons; Nuclear; Numbers; Operative Surgical Procedures; Outcome; Outcome Measure; Pathology; Pathway interactions; Play; Principal Investigator; Property; Protein Overexpression; Proteins; Proteolysis; Proteomics; Recovery; Relative (related person); Research Personnel; Rodent; Role; Serum; Signal Pathway; Signaling Protein; Silver Staining; Solubility; Specificity; Spectrin; Staining method; Stains; Standardization; System; Techniques; Testing; Therapeutic; Time; Tissue Sample; Tissues; Transgenes; Translating; Translations; Trauma; Traumatic Brain Injury; Weight; apoptosis inducing factor; base; brain tissue; calpain inhibitor; calpastatin; clinically relevant; controlled cortical impact; dentate gyrus; endonuclease G; functional disability; functional improvement; improved; in vivo; inhibitor/antagonist; injured; innovation; insight; memory retention; neurofilament; neuron loss; neuronal survival; neuroprotection; novel; novel therapeutics; pre-clinical; prevent; programs; research study; response; small molecule; tau Proteins; therapeutic target; tool

Traumatic brain injury (TBI) results in the prolonged activation of calpains, which contributes to cytoskeletal damage, neuronal death and behavioral dysfunction. However, surprisingly few in vivo cellular substrates of calpains have been identified in the traumatically injured brain and, consequently, little is understood about the pathways through which calpains mediate posttraumatic morbidity. Calpastatin, the endogenous inhibitor of calpains, is the only known protein that exclusively inhibits calpains. As such, calpastatin represents an ideal molecular tool with which to isolate the actions of calpains within the injured brain. Efforts to translate exciting preclinical data demonstrating functional improvement in brain-injured rodents treated with exogenous calpain inhibitors have been slowed by challenges with solubility, specificity and bioavailability of small molecule inhibitors. Enhancing endogenous calpastatin activity may represent a novel and potent therapeutic approach. The overall goals of Project 1, then, are to evaluate the role of calpastatin in regulating posttraumatic calpain-mediated proteolysis and to assess the neuroprotective and behavioral efficacy of increasing calpastatin activity in the setting of TBI. Using genetically altered mice that either overexpress human calpastatin or are calpastatin deficient, Project 1 will: 1) evaluate the role of calpastatin in modulating behavioral outcome following focal or diffuse brain injury, 2) quantify the effects of altered calpastatin expression on neuronal survival and axonal injury after focal or diffuse brain injury, 3) determine the in vivo role of calpastatin in limiting trauma-induced proteolysis of neuronal cytoskeletal proteins, and 4) evaluate the role of posttraumatic calpain activation in modifying membrane proteins involved in calcium influx and in modulating mitochondria-related cell death events. Our central hypothesis is that calpastatin overexpression will prevent calpain-mediated cleavage of neuronal substrates critical for cell survival, thereby attenuating posttraumatic neuronal death and dysfunction. The proposed experiments will provide the first evidence for a functional role for calpastatin in posttraumatic pathology and elucidate differential roles for the calpain/calpastatin system in focal and diffuse TBI. In addition, this Project will provide the groundwork for novel therapeutic approaches, based on manipulation of the calpastatin system, aimed at attenuating brain damage and dysfunction due to TBI as well as other CMS injury and disease states.

创伤性脑损伤 (TBI) 导致钙蛋白酶延长激活，这有助于细胞骨架 损伤、神经元死亡和行为功能障碍。然而，令人惊讶的是，体内细胞底物很少 钙蛋白酶已在创伤性损伤的大脑中被发现，因此，人们对它知之甚少。 钙蛋白酶介导创伤后发病的途径。 Calpastatin，内源性抑制剂 钙蛋白酶，是唯一已知的专门抑制钙蛋白酶的蛋白质。因此，钙蛋白酶抑制剂代表 分离受损大脑内钙蛋白酶作用的理想分子工具。努力翻译 令人兴奋的临床前数据表明，经过治疗的脑损伤啮齿动物的功能得到改善 由于溶解度、特异性和生物利用度方面的挑战，外源性钙蛋白酶抑制剂的发展速度已经放缓。 小分子抑制剂。增强内源性钙蛋白酶抑制素活性可能代表一种新颖且有效的方法 治疗方法。那么，项目 1 的总体目标是评估钙蛋白酶抑制剂在 调节创伤后钙蛋白酶介导的蛋白水解并评估神经保护和行为 在 TBI 情况下增加 Calpastatin 活性的功效。使用基因改造的小鼠 人类钙蛋白酶抑制素过度表达或钙蛋白酶抑制素缺乏，项目 1 将：1) 评估钙蛋白酶抑制素的作用 在调节局灶性或弥漫性脑损伤后的行为结果时，2）量化改变的影响 局灶性或弥漫性脑损伤后钙蛋白酶表达对神经元存活和轴突损伤的影响，3) 确定 Calpastatin 在限制创伤诱导的神经元细胞骨架蛋白水解中的体内作用，以及 4) 评估创伤后钙蛋白酶激活在修饰涉及钙的膜蛋白中的作用 流入并调节线粒体相关的细胞死亡事件。我们的中心假设是钙蛋白酶抑制剂 过度表达将阻止钙蛋白酶介导的对细胞存活至关重要的神经元底物的裂解， 从而减轻创伤后神经元死亡和功能障碍。拟议的实验将提供 第一个证据表明卡帕他汀在创伤后病理学中具有功能性作用并阐明了差异 钙蛋白酶/钙蛋白酶抑制素系统在局灶性和弥漫性 TBI 中的作用。此外，该项目将提供 基于钙蛋白酶抑制素系统操纵的新型治疗方法的基础，旨在 减轻由 TBI 以及其他 CMS 损伤和疾病状态引起的脑损伤和功能障碍。