Molecular mechanism and physiological function of mitochondrial calcium regulation

线粒体钙调节的分子机制及生理功能

基本信息

批准号：
10192800
负责人：
Julia Chang Liu
金额：
$ 24.9万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10192800
关键词：
Academia Address Advisory Committees Aging Alleles Alzheimer&apos s disease model Animal Model Animals Architecture Ataxia Award Biochemical Genetics Bioenergetics Bioinformatics Biology Calcium Cell Death Cell Line Cell physiology Cells Clinical Collaborations Communication Complex Computer Analysis Core Facility Cultured Cells Data Defect Development Plans Disease Disease Progression Embryonic Development Ensure Environment Exhibits Fellowship Gatekeeping Gene Expression Generations Genes Genetic Goals Grant Homeostasis Human In Vitro Job Application Knock-out Lead Learning Longevity Mass Spectrum Analysis Mediating Mediation Membrane Proteins Memory Mentors Mentorship Metabolism Methods Mitochondria Modeling Molecular Morphology Multiprotein Complexes Mus Muscle Muscle Weakness Muscular Dystrophies Mutation Myopathy National Heart, Lung, and Blood Institute Neurodegenerative Disorders Neurologic Dysfunctions Oral Pathology Patients Perinatal mortality demographics Phenotype Physiological Physiology Play Positioning Attribute Precipitation Production Professional Competence Proteins Proteomics Publications Reagent Regulation Reperfusion Injury Reporting Research Rest Role Scientist Secure Signal Pathway Signal Transduction Stress Symptoms Techniques Testing Therapeutic Tissues Training Work age related aged calcium uniporter career development design experience gel electrophoresis genetic approach in vivo knock-down loss of function mitochondrial permeability transition pore mouse model muscle strength mutant nervous system disorder normal aging novel reconstitution scaffold skills stem cell homeostasis symptomatic improvement tenure track transcriptome transcriptome sequencing uptake

项目摘要

Project Summary/Abstract The candidate for this award seeks further mentored research experience while developing career skills to facilitate a successful transition to research independence. Therefore, she has proposed additional experimental and professional training during her postdoctoral fellowship in the lab of Dr. Toren Finkel at the NHLBI. She will take advantage of the outstanding scientific environment in her mentor's lab and at the NHLBI to learn new techniques, including proteomics, RNA sequencing, bioinformatics and computational analysis, and animal work including advanced phenotyping tests. To train her in these methods and assist her in her research aims, she has established collaborations with Dr. Elizabeth Murphy's lab and several NHLBI core facilities. Furthermore, the candidate has designed a career development plan to ensure she prepares thoroughly for an academic position by cultivating her oral and written communication, mentorship, and lab management skills. The candidate has assembled an advisory committee consisting of her primary mentor and several other scientists who not only have extensive scientific experience in fields related to the mitochondrial biology proposed in this grant, but also have committed to guiding her on presentations, job applications, and negotiation strategies. This training will help the candidate secure a tenure-track position in academia. The research goal of this proposal is to dissect the molecular mechanism and physiological role of mitochondrial calcium regulation. Mitochondrial uptake of calcium can help to stimulate ATP production, but too much calcium can lead to opening of the mitochondrial permeability transition pore, triggering cell death. The selective channel through which calcium can rapidly enter the mitochondria, the mitochondrial calcium uniporter, is a multi-protein complex whose components are beginning to be identified. EMRE and MICU1 are two of these proteins that in cell lines have been shown to play critical roles in regulation of calcium uptake. The candidate has generated the first mouse models of EMRE and MICU1 deletion to elucidate the in vivo role of mitochondrial calcium. Her recent publication showed that MICU1 deletion leads to mitochondrial calcium overload, leading to drastically decreased survival and other defects. She will next characterize the effect of EMRE deletion on the molecular architecture of the uniporter as well as on organismal physiology (Aim 1). Her preliminary data suggest that without EMRE, mitochondria cannot uptake calcium. Therefore, the candidate will use MICU1 and EMRE deletion as genetic reagents representing “gain” and “loss” of function in terms of mitochondrial calcium uptake to elucidate how mitochondrial calcium regulation alters in vivo physiological function in global gene expression, aging, and disease (Aim 2). A number of muscular and neurodegenerative diseases have long been associated with mitochondrial calcium overload, but the generation of these mouse models will enable the first direct tests of the impact of mitochondrial calcium on organismal physiology. The completion of these aims thus is of both basic and clinical importance.

项目概要/摘要该奖项的候选人寻求导师进一步的研究经验，同时发展职业技能因此，她提出了额外的建议。在托伦·芬克尔博士的实验室进行博士后研究期间，她接受了实验和专业培训她将利用导师实验室和 NHLBI 出色的科学环境。学习新技术，包括蛋白质组学、RNA测序、生物信息学和计算分析，和动物工作，包括高级表型测试，培训她使用这些方法并协助她。为了实现研究目标，她与 Elizabeth Murphy 博士的实验室和多个 NHLBI 核心建立了合作此外，候选人还设计了职业发展计划，以确保她做好准备。通过培养她的口头和书面沟通、指导和实验室能力，彻底获得学术职位候选人组建了一个咨询委员会，由她的主要导师和其他几位科学家不仅在线粒体相关领域拥有丰富的科学经验生物学在这笔赠款中提出，但也致力于指导她进行演讲、工作申请和该培训将帮助候选人在学术界获得终身职位。本提案的研究目标是剖析其分子机制和生理作用线粒体钙调节。线粒体对钙的吸收也有助于刺激 ATP 的产生。大量的钙会导致线粒体通透性转换孔打开，引发细胞死亡。钙能快速进入线粒体的选择性通道，线粒体钙 uniporter 是一种多蛋白复合物，其成分已开始被鉴定。其中两种蛋白质在细胞系中已被证明在调节钙吸收方面发挥着关键作用。该候选人已生成第一个 EMRE 和 MICU1 缺失的小鼠模型，以阐明其体内作用她最近发表的论文表明，MICU1 缺失会导致线粒体钙减少。超负荷，导致生存率急剧下降和其他缺陷。接下来她将描述其影响。 EMRE 缺失对单向转运蛋白的分子结构以及生物生理学的影响（目标 1）。初步数据表明，如果没有 EMRE，线粒体就无法吸收钙。使用 MICU1 和 EMRE 缺失作为代表功能“获得”和“丧失”的遗传试剂线粒体钙摄取阐明线粒体钙调节如何改变体内生理在整体基因表达、衰老和疾病中发挥作用（目标 2）。长期以来，疾病一直与线粒体钙超载有关，但这些小鼠的产生该模型将首次直接测试线粒体钙对生物生理学的影响。因此，完成这些目标具有基础和临床重要性。