The Role of Intracellular Serpins in the Regulation of Necrosis

细胞内丝氨酸蛋白酶抑制剂在坏死调节中的作用

• 批准号: 8068807
• 负责人: Mark T. Miedel
• 金额: $ 5.13万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8068807
• 关键词: Animals; Biological Models; Caenorhabditis elegans; Cell Death; Chronic; Development; Disease; Gastrointestinal Diseases; Genes; Goals; Hereditary Disease; Homeostasis; Inflammation; Injury; Molecular; Necrosis; Pathologic; Pathway interactions; Patients; Peptide Hydrolases; Process; Proteins; RNA Interference; Regulation; Research Proposals; Role; Serpins; Signal Recognition Particle; Stimulus; Stress; base; insight; positional cloning; public health relevance; research study; response; therapeutic development

DESCRIPTION (provided by applicant): Necrosis is a common pathologic feature of numerous diseases, but the molecular details regarding the execution of this pathway remain largely unknown. Our lab has produced significant insight into the mechanisms involved in necrotic cell death examining the anti-protease function of intracellular serpin proteins. Using the C. elegans model system, it has been shown that SRP-6 functions to block necrosis in response to multiple stress-inducing stimuli. More recent studies demonstrate that lack of SRP-6 results in impaired protein homeostasis, and that increased proteotoxicity results in necrotic cell death in either wild- type or srp-6 null animals. The long term objectives of this project are to dissect the relationship between proteotoxicity and necrotic cell death, and to identify specific genes involved in the regulation of these processes. The C. elegans model system will be used to examine the following specific aims: 1) to determine how the loss of SRP-6 perturbs protein homeostasis and 2) to determine how perturbed protein homeostasis results in necrotic cell death. In Aim 1 I will perform rescue experiments in srp-6 null animals to determine whether SRP-6 inhibitory activity is involved in the regulation of protein homeostasis. Additionally, I will employ an RNAi screen to identify specific peptidases as SRP-6 regulatory targets. In Aim 2, I will utilize both forward and reverse genetic approaches to identify specific genes that modulate the protein homeostasis and/or NCD pathways. The results of these studies will provide new insights into the mechanisms that regulate protein homeostasis and necrosis, facilitating the development of therapeutic options for treatment of necrosis-related diseases. PUBLIC HEALTH RELEVANCE: Necrosis is a common pathologic feature of many diseases including those associated with ischemic injury, chronic inflammation, and many gastrointestinal disorders. The goal of this research proposal is to determine the molecular mechanisms and to identify critical genes that contribute to necrotic cell death. Ultimately, these studies will provide the basis for development of highly specific treatments for patients with necrosis-associated disorders.

描述（由申请人提供）：坏死是多种疾病的常见病理特征，但是有关该途径执行的分子​​细节仍然在很大程度上未知。我们的实验室已经对参与坏死细胞死亡的机制进行了重大洞察力，研究了细胞内SERPIN蛋白的抗蛋白酶功能。使用秀丽隐杆线虫模型系统，已经表明SRP-6的功能可以响应多种应激刺激而阻塞坏死。最近的研究表明，缺乏SRP-6导致蛋白质稳态受损，并且增加蛋白质毒性会导致野生型或SRP-6无效动物的坏死细胞死亡。该项目的长期目标是剖析蛋白毒性与坏死细胞死亡之间的关系，并确定与这些过程调节有关的特定基因。秀丽隐杆线虫模型系统将用于检查以下特定目的：1）确定SRP-6 Perturbs蛋白稳态的丢失和2）如何确定扰动蛋白稳态如何导致坏死细胞死亡。在AIM 1中，我将在SRP-6无效动物中进行救援实验，以确定SRP-6抑制活性是否参与了蛋白质稳态的调节。此外，我将采用RNAi筛选来鉴定特定的肽酶为SRP-6调节靶标。在AIM 2中，我将利用前进和反向遗传方法来识别调节蛋白质稳态和/或NCD途径的特定基因。这些研究的结果将为调节蛋白质稳态和坏死的机制提供新的见解，从而促进治疗与坏死相关疾病的治疗方案的发展。 公共卫生相关性：坏死是许多疾病的常见病理特征，包括与缺血性损伤，慢性炎症和许多胃肠道疾病有关的疾病。该研究建议的目的是确定分子机制，并确定导致坏死细胞死亡的关键基因。最终，这些研究将为与坏死相关疾病患者开发高度特定治疗的基础。