Caspase-independent cell death and neurodegeneration

不依赖半胱天冬酶的细胞死亡和神经变性

• 批准号: 6465018
• 负责人: ALEXEI DEGTEREV
• 金额: $ 11.88万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6465018
• 关键词: Parkinson's disease; apoptosis; calcium disorder; cell biology; cell death; cell growth regulation; cell population study; cysteine endopeptidases; disease /disorder model; flow cytometry; gene expression; genetic regulation; ischemia; laboratory mouse; mitochondrial disease /disorder; molecular biology; molecular pathology; necrosis; neural degeneration; neurons; oxidative stress; postdoctoral investigator; tissue /cell culture

DESCRIPTION (provided by applicant): The overall goal of this project is to characterize a novel cell death pathway, termed "aponecrosis", and determine its role in neurodegeneration. Extensive evidence was generated in recent years suggesting that two "classic" cell death pathways, apoptosis and necrosis, do not explain the variety of physiological and pathological cell death mechanisms. Existence of the third pathway, termed "aponecrosis", is proposed. This pathway is activated in cells that are induced to undergo apoptosis, yet prevented from its completion. It is proposed to represent a novel safety mechanism aimed at elimination of damaged and potentially dangerous cells. Aponecrosis shares programmed cell death nature with apoptosis and execution subroutines and phenotypic features with necrosis. Using high throughput screening of small molecule library several chemical inhibitors of aponecrosis were isolated. These compounds were found to selectively inhibit aponecrosis, but not apoptosis, underscoring distinct mechanism of the aponecrotic cell death process. Using selected inhibitors aponecrosis was shown to represent a major cell death process in a variety of caspase-independent cell death paradigms in vitro. Aponecrosis was also implicated in amyloid-beta toxicity in PC12 cells and ischemic brain damage in vivo. Analysis of the effects of selected inhibitors in various systems suggests that two of them target cell type specific signaling pathways, whereas another compound blocks uniform downstream execution step, providing the opportunity to characterize various steps in aponecrosis with the help of individual inhibitors. Two aponecrotic regulators (p38 kinase and cathepsin B) were discovered. Specific aims of this study are as follows: 1 ) To evaluate the role of aponecrosis in neuronal cell death in vitro and in vivo; 2) To characterize molecular events involved in aponecrosis; 3) To identify novel genes involved in aponecrosis using chemical, cell and molecular biology approaches. The research will be conducted in the Department of Cell Biology at Harvard Medical School under the joint guidance of Dr. Junying Yuan and Dr. Timothy Mitchison. Dr. Junying Yuan is a recognized leader in the field of neuronal cell death. Dr. Timothy Mitchison is an outstanding cell biologist, who is also one of the pioneers of chemical biology approach. Dr. Yuan will provide resources and mentorship for the first two and partially for the third aim. Dr. Timothy Mitchison will provide training and supervision as well as resources of Harvard Institute of Chemistry and Cell Biology for the chemical biology part of the project. Department of Cell Biology is fully committed to the applicant's development into a fully independent researcher in the field of aging-related neurodegeneration. Dr. Alexei Degterev is aspiring to become a fully independent research scientist in the field of aging-associated neuronal cell death and would use K01 training period to develop and enhance the skills needed to perform independent research in this area and develop research project that will from the basis for his future independent research.

描述（申请人提供）：本项目的总体目标是 表征一种新的细胞死亡途径，称为“坏死”，并确定 它在神经退化中的作用最近有大量证据表明 多年来，这表明两个“经典”的细胞死亡途径，凋亡和 坏死，不说明生理和病理细胞的变化 死亡机制第三种途径的存在，被称为“腱膜坏死”， 提出了这种途径在细胞中被激活， 凋亡，但阻止其完成。建议代表A 新的安全机制，旨在消除损坏和潜在的 危险的细胞腱膜坏死与 细胞凋亡和执行子程序和坏死的表型特征。 利用高通量筛选小分子库的几种化学物质 分离出腱膜坏死的抑制剂。发现这些化合物 选择性抑制腱膜坏死，但不抑制凋亡，强调了不同的 腱膜坏死细胞死亡过程的机制。使用选定的抑制剂 腹膜坏死显示代表了多种肿瘤中主要的细胞死亡过程， 半胱天冬酶非依赖性细胞死亡模式。腱膜坏死也是 与PC 12细胞中的β淀粉样蛋白毒性和缺血性脑损伤有关。 vivo.分析所选抑制剂在各种体系中的作用 表明其中两种靶向细胞类型特异性信号通路， 而另一种化合物阻止统一的下游执行步骤， 有机会在腱膜坏死的帮助下， 个别抑制剂。两种脂膜坏死调节因子（p38激酶和组织蛋白酶B） 被发现了本研究的具体目的如下：1）评价 在体外和体内的神经元细胞死亡中，腱膜坏死的作用; 2） 表征涉及腱膜坏死的分子事件; 3）鉴定新的 利用化学、细胞和分子生物学研究与腱膜坏死有关的基因 接近。 这项研究将在哈佛的细胞生物学系进行 在袁俊英博士和蒂莫西博士的共同指导下， 米奇森袁俊英博士是神经元领域公认的领导者， 细胞死亡Timothy Mitchison博士是一位杰出的细胞生物学家， 也是化学生物学方法的先驱之一。袁医生将提供 为前两个目标提供资源和指导，部分用于第三个目标。 Timothy Mitchison博士将提供培训和监督， 哈佛化学与细胞生物学研究所的化学资源 项目的生物学部分。细胞生物学系完全致力于 申请人发展成为该领域完全独立的研究人员 与衰老有关的神经退化 阿列克谢·德格捷列夫博士渴望成为一个完全独立的研究 衰老相关神经细胞死亡领域的科学家，并将使用 K 01培训期，以发展和提高执行所需的技能 在这一领域进行独立研究，并开发研究项目， 这是他未来独立研究的基础。