Mitochondrial calcium homeostasis in SOD1-familial ALS

SOD1 家族性 ALS 中的线粒体钙稳态

• 批准号: 8075416
• 负责人: Giovanni Manfredi
• 金额: $ 36.23万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8075416
• 关键词: ATP Synthesis Pathway; Ablation; Adenylate Cyclase; Advocate; Biochemical; Bioenergetics; Buffers; Calcium; Caring; Cell Death; Cessation of life; Chronic; Cyclic AMP; Data; Disease; Epidemiology; Estrogen Receptors; Estrogens; Failure; Female; Fractionation; Genetic; Goals; Homeostasis; Immunoelectron Microscopy; Investigation; Life; Mediating; Mitochondria; Modeling; Molecular; Mus; Mutant Strains Mice; Mutation; Neurons; Oxidative Phosphorylation; Pathogenesis; Pathway interactions; Play; Precipitation; Predisposition; Prevention; Regulation; Role; Safety; Sex Characteristics; Signal Transduction; Testing; Transgenic Mice; UCP2 protein; Wild Type Mouse; Woman; base; cyclophilin D; improved; in vivo; late disease onset; male; man; mitochondrial dysfunction; mouse model; mutant; neuroprotection; novel; novel therapeutics; overexpression; prevent; public health relevance

DESCRIPTION (provided by applicant): This is an application for renewal of the project entitled "the pathogenic role of mutant SOD1 in mitochondria". Mitochondrial Ca2+ overload leads to deleterious consequences, including mitochondrial de-energization, structural changes, and bioenergetic failure, which can result in cell death. The overarching hypothesis of this application is that mitochondrial Ca2+ overload plays a fundamental role in the pathogenesis of familial ALS (FALS) associated with SOD1 mutations. Mitochondrial dysfunction, one of the cardinal features of ALS, causes increased susceptibility to Ca2+ overload. Our preliminary results show impaired mitochondrial Ca2+ handling and susceptibility to overload in the CNS of mutant SOD1 transgenic mice. They also show that female mutant SOD1 mice are partially protected against mitochondrial Ca2+ overload by a pathway of Ca2+ release involving estrogen, mitochondrial estrogen receptor (ER), and cyclophilin D (CyPD), a modulator of mitochondrial Ca2+ induced damage. In this application, we provide a mechanistic hypothesis for the involvement of mitochondria and for the gender differences in FALS. The broad goals are to define the mechanisms of mitochondrial Ca2+ handling regulation and to develop approaches to prevent Ca2+-overload in FALS mitochondria. Our specific aims are to: 1) Define the role of the estrogen receptor in Ca2+ handling in FALS mitochondria by investigating, in vivo and ex vivo, ER2 KO mice crossed with G93A mutant SOD1 mice. Then, with the G85R mutant SOD1 mouse model, to determine if the mechanisms of estrogen-ER2 and CyPD dependent neuroprotection are common to different SOD1 mutants. 2) Define the biochemical and molecular basis of estrogen-ER regulation of mitochondrial Ca2+ handling and the effects of mutant SOD1. In the mouse crosses established in aim 1, we will determine: i) how estrogen-ER2 modulates mitochondrial bioenergetics and the CyPD- dependent Ca2+ release pathway, ii) the mitochondrial localization of the ER2 and the ER- CyPD interactions with SOD1, by mitochondrial fractionation, immuno-electron microscopy, and immuno-precipitation. 3) Test different complementary approaches to prevent Ca2+ overload in FALS mitochondria and improve disease in mutant SOD1 mice by: i) chronic administration of estrogen to male SOD1 mutant mice, ii) boosting mitochondrial bioenergetics with expression of a mitochondrial soluble adenylyl cyclase that enhances oxidative phosphorylation, iii) mild mitochondrial uncoupling achieved with overexpression of uncoupling protein 2 (UCP2). PUBLIC HEALTH RELEVANCE: Mitochondria play a fundamental role in regulating neuronal life and death. One of the main functions of mitochondria is to take care of intracellular calcium (Ca2+). Mitochondrial dysfunction is one of the cardinal features of ALS and causes increased susceptibility to Ca2+ overload. Mitochondrial Ca2+ overload leads to deleterious consequences, including mitochondrial failure, which can result in cell death. The leading hypothesis of this study is that mitochondrial Ca2+ overload plays a fundamental role in the pathogenesis of familial ALS (FALS) associated with SOD1 mutations. Therefore the broad goals of this project are to define the mechanisms of mitochondrial Ca2+ handling regulation and to develop approaches to prevent Ca2+-overload in FALS mitochondria to ameliorate the disease.

描述（由申请人提供）：这是续签“突变SOD1在线粒体中的致病作用”的申请。线粒体Ca2+过载会导致有害后果，包括线粒体去启动，结构变化和生物能衰竭，这可能导致细胞死亡。该应用的总体假设是线粒体Ca2+过载在与SOD1突变相关的家族性ALS（FAL）的发病机理中起着基本作用。线粒体功能障碍是ALS的主要特征之一，导致对Ca2+过载的敏感性增加。我们的初步结果表明，线粒体Ca2+处理和易感性在突变SOD1转基因小鼠的中枢神经系统中过载。他们还表明，通过涉及雌激素，线粒体雌激素受体（ER）的Ca2+释放途径，雌性突变SOD1小鼠被部分保护，以防止线粒体Ca2+过载，并释放了环蛋白D（CYPD）（CYPD），这是线粒体CA2+诱导的损伤的调节剂。 在此应用中，我们为线粒体的参与和伪造的性别差异提供了一种机械假设。广泛的目标是定义线粒体Ca2+处理调节的机制，并开发方法以防止fals线粒体中的Ca2+超载。我们的具体目的是：1）定义雌激素受体在Ca2+处理中通过研究与G93A突变体Sod1小鼠交叉的ER2 KO小鼠在Ca2+处理中的作用。然后，使用G85R突变体SOD1小鼠模型，以确定雌激素-ER2和CYPD依赖性神经保护的机制是否对不同的SOD1突变体是常见的。 2）定义线粒体Ca2+处理的雌激素-ER调节的生化和分子基础和突变SOD1的效果。在AIM 1中建立的小鼠杂交中，我们将确定：i）雌激素-ER2如何调节线粒体生物能学和依赖性CA2+释放途径，ii）II）通过线粒体交易，Immoccation，Immoscrion，Immoscry-ectron，Immoscrion，Immoscron，Immoscron，Immoscron，Immoscry-ectron，Immoscrion，Immoscron，Immoscron，Immoscron and ectron and croproproy，Immoscron and cron and ectron。 3）测试不同的互补方法，以防止通过以下方式通过以下方式进行线粒体中的Ca2+过载并改善突变体Sod1小鼠的疾病，i）长期将雌激素慢性施用对雄性SOD1突变小鼠，ii）促进线粒体生物生物生物生物学的促进，以增强线粒体溶解度的氧化粘液酶的表达，从而增强了氧化均匀环境。解偶联蛋白2（UCP2）的过表达实现。 公共卫生相关性：线粒体在调节神经元的生命和死亡方面起着基本作用。线粒体的主要功能之一是照顾细胞内钙（Ca2+）。线粒体功能障碍是ALS的基本特征之一，导致对CA2+过载的敏感性增加。线粒体Ca2+过载会导致有害后果，包括线粒体衰竭，这可能导致细胞死亡。这项研究的主要假设是，线粒体Ca2+超负荷在与SOD1突变相关的家族性ALS（FAL）的发病机理中起着基本作用。因此，该项目的广泛目标是定义线粒体Ca2+处理调节的机制，并开发方法以防止通过fals fals线粒体改善疾病的Ca2+误解。