Effects of environmental stressors on mitochondrial-cellular cross-talk

环境压力源对线粒体细胞串扰的影响

基本信息

批准号：
9058544
负责人：
NORBERT PERRIMON
金额：
$ 20.28万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-05-01 至 2017-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9058544
关键词：
Address Adipose tissue Adult Affect Applications Grants Biological Markers Cadmium Cell Nucleus Citrate (si)-Synthase Clustered Regularly Interspaced Short Palindromic Repeats Complex Constitution Consumption Dose Drosophila genus Engineering Event Exposure to Gene Expression Genes Genome Goals Grant Health Label Life Measures Mercury Metals Mitochondria Mitochondrial Proteins Molecular Muscle Nuclear Ontology Oxygen Consumption Pathway Analysis Pathway interactions Peroxidases Phase Proteins Proteome Proteomics Research Signal Transduction Stress System Techniques Time Tissues Toxic Environmental Substances Zinc ascorbate base biological adaptation to stress brain tissue cell type environmental stressor feeding fly inhibitor/antagonist novel protein complex response sensor stressor tool toxic metal toxicant transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): This R21 grant application grant application addresses the goals stated in the Research Objectives. Specifically, the intent is to establish a robust platform in Drosophila tissues to characterize changes occurring in mitochondria (activity, protein composition), and in the genome (nuclear-encoded mitochondrial genes and others), in response to exposure to environmental mitochondrial stressors, such as toxic metals. The initial studies will focus on the gut, as it is the tissue where metals are first absorbed. Results from these studies will allow one to engineer flies carrying biomarkers of environmental mitochondrial toxicants that can be used to dissect the molecular mechanisms underlying cross-talk and signaling between mitochondria and the nucleus. The grant application is composed of three specific aims: Specific Aim 1) Characterize changes in mitochondrial activities and proteome constitution in response to environmental mitochondrial toxicants in the adult Drosophila gut. Mitochondrial activities, including citrate synthase activity and O2 consumption rate, as well as mitochondrial proteome constitution, will be measured at different timepoints following toxic metals feedings. For the proteomic studies, a novel protein labeling technique will be exploited, based on an engineered ascorbate peroxidase (APEX), that was recently adapted for studies in Drosophila live tissues; Specific Aim 2) Characterize the transcriptional changes occurring in the Drosophila gut in response to environmental mitochondrial toxicants. RNA-Seq of gut will be examined at different timepoints to characterize the transcriptional changes occurring in this tissue following feeding of toxic metals. Further, using the recently developed COMPLEAT tool, mitochondrial protein complexes that are preferentially affected by environmental mitochondrial toxicants will be determined; and Specific Aim 3) Generate biomarkers of environmental mitochondrial stress. Genes will be selected that are induced by environmental mitochondrial stressors to engineer, using CRISPR, GFP sensors that can be used to dissect the mitochondrial toxicant response. Following the R21 phase, we envision expanding this project into an R33 where we will: 1) Study additional environmental mitochondrial stressors to establish signatures of specific toxicants; 2) Expand to other tissues (brain, muscle, and adipose tissues) to gain a general understanding of the diversity of responses of different cell types; and 3) Use engineered flies carrying biomarkers of environmental mitochondrial toxicants to dissect the molecular mechanisms underlying cross-talk and signaling between mitochondria and the nucleus.

描述（由应用程序提供）：此R21授予应用程序授予应用程序解决了研究目标中所述的目标。具体而言，目的是在果蝇组织中建立一个健壮的平台，以表征线粒体（活性，蛋白质成分）和基因组（核编码的线粒体基因等）中发生的变化，以响应于环境线粒体压力的暴露，例如毒性金属。最初的研究将集中在肠道上，因为它是首先吸收金属的组织。这些研究的结果将使人们能够设计携带环境线粒体毒物的生物标志物，这些果实可用于剖析串扰和线粒体之间的串扰和信号传导的分子机制。该赠款的应用由三个特定目的组成：特定目的1）表征线粒体活性的变化和蛋白质组线粒体活性（包括柠檬酸盐合酶活性和O2的消耗率，以及线粒体蛋白质成分），将在毒性金属喂养后的不同时间点上测量。对于蛋白质组学研究，将基于工程的抗坏血酸过氧化物酶（APEX）探索一种新型的蛋白质标记技术，该技术最近适用于果蝇活组织中的研究。具体目标2）表征果蝇肠道中发生的转录变化，以响应环境线粒体毒物。将在不同的时间点上检查肠道的RNA-seq，以表征有毒金属进食后这种组织中发生的转录变化。此外，使用最近开发的完整工具，将确定更可能受环境线粒体毒物影响的线粒体蛋白质复合物。和特定目的3）产生环境线粒体应力的生物标志物。将使用CRISPR，GFP传感器来选择由环境线粒体压力诱导的基因，可用于剖析线粒体有毒反应。在R21阶段之后，我们将该项目扩展到R33的环境：1）研究其他环境线粒体应力源以建立特定毒物的特定签名； 2）扩展到其他组织（大脑，肌肉和脂肪组织），以对不同细胞类型的反应多样性有一般的了解；和 3）使用携带环境线粒体有毒物质生物标志物的工程苍蝇，以剖析线粒体和细胞核之间的串扰和信号传导的分子机制。