Zone-specific mitochondrial functions in regulation of hepatic metabolism

区域特异性线粒体功能在肝代谢调节中的作用

基本信息

批准号：
10788519
负责人：
CHRISTOPHER B NEWGARD
金额：
$ 44.28万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-30 至 2025-09-29
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10788519
关键词：
Affect Age Aging Albumins Amino Acids Anatomy Area Biochemical Biological Assay CRISPR screen Candidate Disease Gene Cell Nucleus Cells Cirrhosis Clonal Expansion Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Coupled DNA sequencing Development Diet Disease Engineering Epitopes Eukaryotic Cell Evaluation Exhibits Exploratory/Developmental Grant Fibrosis Functional disorder Genes Genomic DNA Glucose Goals Guide RNA Health Hepatic Hepatocellular Damage Hepatocyte Heterogeneity Homeostasis Image Impairment Incidence Inflammation Label Laboratories Lipids Liver Liver Dysfunction Liver Failure Liver Mitochondria Liver diseases Lobule Measurement Metabolic Metabolic Diseases Metabolic Pathway Metabolic stress Metabolism Methods Mitochondria Modernization Molecular Mouse Strains Mus Mutate Nature Nutritional Obesity Overnutrition Pathogenesis Patients Personal Satisfaction Physiology Play Population Prevalence Prevention Primary carcinoma of the liver cells Progressive Disease Property Proteins Proteomics Quality of life Radial Regulation Respiratory physiology Risk Role Site Stress Syndrome Technology Time Tissues Transgenic Mice United States Work aged aging nutrition design fitness gene discovery in vivo innovative technologies liver metabolism metabolomics middle age mitochondrial dysfunction mitochondrial metabolism mortality non-alcoholic fatty liver disease nonalcoholic steatohepatitis novel novel therapeutics pandemic disease respiratory response screening selective expression tool transcriptomics

项目摘要

.SUMMARY – R21 – Newgard . Aging and overnutrition have deleterious effects on liver health, including development of mitochondrial dysfunction. The liver is divided into distinct zones (Zone 1–Zone 3), each enriched in a discrete subset of metabolic pathways, but little is known about how aging and metabolic stress affect zone-specific mitochondrial metabolic or respiratory functions. Using a powerful suite of technologies, including mice engineered for zone- specific expression of an immunoisolation tag (MITO-Tag) that allows purification of mitochondria in <10 minutes, sophisticated metabolomic and metabolic flux analysis tools, mitochondrial respiratory assays, and in vivo CRISPR gene discovery technologies, we will (i) optimize our workflow for use of MITO-Tag for rapid isolation of mitochondria from liver at high yield and with full metabolic functionality (Aim 1); ii) develop zone- specific MITO-Tag mice to allow a comprehensive view of metabolism and energetics in mitochondria isolated from different liver zones (Aim 2); (iii) deploy two in vivo CRISPR screening approaches, including a novel method designed in our laboratory, to identify mitochondrial genes that impact hepatocellular fitness during nutritional stress (Aim 3). By understanding exactly how metabolism is partitioned across zones in primary liver cells, and by identifying genes via our CRISPR screens that enhance the well-being of liver cells in each zone, we can contribute to development of novel therapies for the growing pandemic of liver dysfunction in the rapidly aging US population.

.Summary - R21 - Newgard。衰老和营养不良已删除对肝脏健康的影响，包括线粒体的发展功能障碍。肝脏分为不同的区域（区域3区），每个区域都富含一个离散的子集代谢途径，但对衰老和代谢应力如何影响区域特异性线粒体知之甚少代谢或呼吸功能。使用强大的技术套件，包括针对区域设计的小鼠免疫分散标签（mito-tag）的特定表达，该标签允许在<10中纯化线粒体分钟，复杂的代谢组和代谢通量分析工具，线粒体呼吸测定和在体内CRISPR基因发现技术，我们将（i）优化我们的工作流程用于快速使用mito-tag 高产率从肝脏中分离线粒体并具有完全代谢功能（AIM 1）； ii）开发区 - 特定的Mito-Tag小鼠可以全面了解线粒体中的新陈代谢和能量学来自不同的肝区（AIM 2）；（iii）部署两种体内CRISPR筛选方法，包括一种小说在我们的实验室中设计的方法，以识别影响肝细胞适应性的线粒体基因营养压力（AIM 3）。通过准确了解原发性肝脏中区域的新陈代谢如何分裂细胞，以及通过我们的CRISPR筛选鉴定基因，从而增强每个区域中肝细胞的福祉，我们可以为迅速发展的肝功能障碍日益增长的新疗法发展做出贡献美国人口老龄化。