The Role of Intracellular Serpins in the Regulation of Necrosis

描述（由申请人提供）：坏死是许多疾病的常见病理特征，但关于该途径的执行的分子细节在很大程度上仍然未知。我们的实验室已经产生了显着的洞察机制参与坏死细胞死亡检查细胞内丝氨酸蛋白酶抑制剂蛋白的抗蛋白酶功能。利用C.在elegans模型系统中，已经显示SRP-6响应于多种应激诱导刺激而起到阻断坏死的作用。最近的研究表明，缺乏SRP-6会导致蛋白质稳态受损，并且蛋白毒性增加会导致野生型或SRP-6缺失动物的坏死性细胞死亡.这个项目的长期目标是剖析蛋白毒性和坏死细胞死亡之间的关系，并确定参与这些过程的调控的特定基因。梭elegans模型系统将用于检查以下特定目的：1）确定SRP-6的丧失如何扰乱蛋白质稳态和2）确定扰乱的蛋白质稳态如何导致坏死细胞死亡。在目的1中，我将在srp-6缺失动物中进行拯救实验，以确定SRP-6抑制活性是否参与蛋白质稳态的调节。此外，我将采用RNAi筛选来鉴定作为SRP-6调节靶点的特定肽酶。在目标2中，我将利用正向和反向遗传学方法来鉴定调节蛋白质稳态和/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，促进治疗坏死相关疾病的治疗选择的发展。 公共卫生相关性：坏死是许多疾病的共同病理特征，包括与缺血性损伤、慢性炎症和许多胃肠道疾病相关的疾病。这项研究计划的目标是确定分子机制，并确定导致坏死细胞死亡的关键基因。最终，这些研究将为坏死相关疾病患者的高度特异性治疗提供基础。

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.