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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.