CPLA2 MEDIATED CELL DEATH AND BC1-2 REGULATION

CPLA2 介导的细胞死亡和 BC1-2 调节

• 批准号: 6380044
• 负责人: ADAM SAPIRSTEIN
• 金额: $ 12.46万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-15 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6380044
• 关键词: DNA footprinting; apoptosis; arachidonate; cell death; cellular pathology; enzyme activity; epithelium; gene targeting; genetic regulation; genetic transcription; genetically modified animals; hydrogen peroxide; immunofluorescence technique; kidney cell; laboratory mouse; molecular cloning; nuclear membrane; oxidative stress; oxidizing agents; phospholipase A2; protooncogene; renal ischemia /hypoxia

The candidate, Adam Sapirstein, was trained in anesthesia and critical care medicine at the Massachusetts General Hospital. His clinical interest is in diseases of critical illness and particularly acute renal failure. Since concluding his clinical training in 1994, he has undertaken an intensive research training program in the laboratory of Dr. Joseph V. Bonventre. The long term objective of the candidate is to independently investigate mechanisms of acute renal failure and improve the therapy and outcome for patients with this disease. Increased phospholipase activity has been implicated in cellular injury and death in many conditions including: ischemia and reperfusion, oxidant stress, inflammation and sepsis. This project~s broad objectives are to determine mechanisms of potentiation of injury induced by cytosolic phospholipase A2 (cPLA2) and to evaluate how cells protect themselves from cPLA2 - mediated injury. There mechanisms of injury and protection will yield important insights into general mechanisms of cell death. The specific aims proposed to achieve them are: Specific Aim 1. To determine if nuclear membrane disruption and enzyme activation are the mechanisms for cPLA2-mediated potentiation of H202- induced cell death in renal epithelia cells. Specific Aim 2. To determine if cPLA2 expression can increase the transcription of the bcl-2 gene in renal epithelial cells through a receptor-mediated gene interaction. To search for other genes that are transcriptionally regulated by cPLA2. Specific Aim 3. To characterize the renal development and response to renal ischemia/reperfusion of a transgenic mouse deficient in cPLA2 (cPLS2 knockout mouse). The methods proposed to achieve the specific aims utilize basic cell and molecular biological techniques that are currently in use in the Bonventre laboratory or in the labs of collaborators. Knowledge of mechanisms of cPLA2-mediate cell injury and cPLA22-regulated gene transcription will lead to important insights into general mechanisms of cell death, and will provide paradigms for prevention and treatment of clinical states in which cPLA2 has been implicated in cell and tissue injury. The environment is the laboratory of Dr. Joseph v. Bonventre at the Massachusetts General Hospital. Dr. Bonventre is an established senior investigator with an interest in acute renal failure who has successfully trained research fellows with a variety of interests. The candidate has established a productive scientific and clinical relationship within the lab and hospital and will be able to utilize the laboratory and institutional resources.

候选人亚当·萨皮尔斯坦接受过麻醉培训， 马萨诸塞州总医院的重症监护医学。 他 临床兴趣在于危重疾病， 急性肾衰竭 自1994年结束临床培训以来， 他在美国进行了一项密集的研究培训计划， Joseph V. Bonventre博士的实验室 的长期目标 候选人将独立研究急性 肾功能衰竭和改善患者的治疗和结果 这种疾病。 磷脂酶活性的增加与 在许多情况下的细胞损伤和死亡，包括： 以及再灌注、氧化应激、炎症和败血症。 这 项目的主要目标是确定 胞浆磷脂酶A2诱导的损伤增强 （cPLA 2），并评估细胞如何保护自己免受cPLA 2- 介导的损伤。 这些损伤和保护机制将 对细胞死亡的一般机制产生重要的见解。 的 为实现这些目标而提出的具体目标是：具体目标1。 到 确定核膜破坏和酶激活 是cPLA 2-介导的H2 O2-增强的机制。 诱导肾上皮细胞死亡。 具体目标2。 到 确定cPLA 2表达是否可以增加细胞的转录， bcl-2基因通过受体介导的基因在肾上皮细胞中的表达 互动 为了寻找其他基因， 由cPLA 2调节。 具体目标3。 表征肾脏 肾缺血/再灌注损伤的发生和反应 cPLA 2缺陷的转基因小鼠（cPLS 2敲除小鼠）。 提出的方法，以实现特定的目标，利用基本细胞 和分子生物学技术，目前正在使用的 Bonventre实验室或合作者的实验室。 知识 cPLA 2介导的细胞损伤和cPLA 22调节的机制 基因转录将带来对一般情况的重要见解 细胞死亡的机制，并将提供范例， 预防和治疗cPLA 2已被 与细胞和组织损伤有关。 环境就是 约瑟夫博士在马萨诸塞州的实验室 医院 Bonventre博士是一位资深研究员， 对急性肾功能衰竭感兴趣， 兴趣广泛的研究员。 这位候选人有 建立了富有成效的科学和临床关系， 实验室和医院，并将能够利用实验室， 机构资源。