Molecular mechanism and physiological function of mitochondrial calcium regulation

项目摘要/摘要 该奖项的候选人寻求进一步指导的研究经验，同时发展职业技能，以 促进向研究独立的成功过渡。因此，她提出了额外的 在托伦·芬克尔博士的实验室担任博士后期间进行的实验和专业培训 NHLBI。她将利用导师实验室和NHLBI的卓越科学环境 为了学习新技术，包括蛋白质组学、RNA测序、生物信息学和计算分析， 以及包括高级表型测试在内的动物工作。对她进行这些方法的培训，并帮助她 研究目标，她与伊丽莎白·墨菲博士的实验室和几个NHLBI核心建立了合作 设施。此外，应聘者还设计了职业发展计划，以确保她为 通过培养她的口头和书面交流、指导和实验室，彻底争取一个学术职位 管理技能。候选人已经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 生物学在这项拨款中提议，但也承诺指导她的演讲，工作申请，和 谈判策略。这一培训将帮助应聘者在学术界获得终身教职。 本课题的研究目的是为了深入研究血管紧张素转换酶的分子机制和生理作用。 线粒体钙调节。线粒体摄取钙有助于刺激三磷酸腺苷的产生，但也 过多的钙会导致线粒体通透性转换孔打开，引发细胞死亡。这个 钙可以快速进入线粒体的选择性通道，即线粒体钙 单转运蛋白，是一种多蛋白质复合体，其组成成分已开始被鉴定。Emre和MICU1是 这些蛋白质中的两种在细胞系中已被证明在钙吸收调节中发挥关键作用。 这位候选人已经建立了第一个EmRE和MICU1缺失的小鼠模型，以阐明在体内的作用 线粒体钙的含量。她最近发表的文章表明，MICU1缺失会导致线粒体钙 过载，导致存活率急剧下降等缺陷。接下来，她将描述 对单转运蛋白的分子结构以及对生物生理学的EmRE缺失(目标1)。她 初步数据表明，如果没有Emre，线粒体就不能吸收钙。因此，候选人将 使用MICU1和EmRE缺失作为遗传试剂，代表功能在以下方面的“获得”和“丧失” 线粒体钙摄取阐明在体生理状态下线粒体钙调节如何改变 在全球基因表达、衰老和疾病中的作用(目标2)。一些肌肉和神经退行性变 长期以来，疾病一直与线粒体钙超载有关，但这些小鼠的后代 模型将使首次直接测试线粒体钙对生物生理的影响成为可能。这个 因此，完成这些目标具有基础和临床重要性。

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.