Illuminating genetic interactions that affect lipid content in adipocytes

阐明影响脂肪细胞脂质含量的遗传相互作用

基本信息

批准号：
10294229
负责人：
Olga Gulyaeva
金额：
$ 2.13万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
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 2021-10-08
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10294229
关键词：
Adipocytes Adipose tissue Adult Affect Alleles Anti-Obesity Agents Appetite Regulation Binding Biogenesis Biological Models CRISPR interference CRISPR library CRISPR screen Caenorhabditis elegans Calories Catabolism Cells Clustered Regularly Interspaced Short Palindromic Repeats Complex Confocal Microscopy Data Set Desire for food Development Diet Disease Dissection Drosophila genus Drug Targeting Effectiveness Epidemic FDA approved Future Gene Expression Regulation Genes Genetic Genome Health Healthcare Human Image Individual Intake Investigation Knowledge Laboratories Lead Left Libraries Life Style Lipids Maps Membrane Metabolic Metabolic Diseases Metabolism Molecular Monitor Morphology Mus Obesity Organ Organelles Overweight Pathologic Pathway interactions Pharmaceutical Preparations Phenotype Physiological Physiology Play Process Proteins Regulation Research Role Saccharomyces cerevisiae Small Interfering RNA System Therapeutic Therapeutic Intervention Time Vesicle adipocyte differentiation arm cell type drug development druggable target energy balance experimental study fascinate functional genomics gene discovery genomic data human disease improved lipid metabolism novel novel therapeutics obese patients obesity treatment pandemic disease programs public health relevance response screening side effect uptake whole genome

项目摘要

Project Abstract Obesity and associated metabolic disorders are a major health care crisis that affect millions of people worldwide and have no long-term treatment. Most of our current knowledge on how cells regulate their metabolism has been obtained using single gene-driven hypothesis. However, obesity like many other diseases is a polygenic disorder, in which multiple genes interact, contributing to phenotype, which makes it impossible to establish the causative allele. If we are to make any headway towards curing obesity and metabolic diseases during this century, we need bold, creative ideas to tackle cellular pathways as oppose to individual genes that take advantage of cutting-edge high-throughput approaches. As of 2016, over 1.9 billion adults worldwide were estimated to be overweight and 650 million adults to be obese. Obesity is classically characterized by pathological development and differentiation of adipose tissue, which is a key energy storage organ in the body. Adipose tissue stores energy in lipid droplets- until recently unappreciated cellular organelles with complex and dynamic functions. Lipid droplets can drastically change in size as a response to changed calorie intake or metabolism. Since currently FDA-approved anti-obesity drugs act on appetite or metabolism regulation and not on adipose tissue lipids directly, we need to identify pathways responsible for lipid droplet biogenesis and breakdown to improve current therapeutics. Surprisingly, very little is known about the pathways that affect lipid droplets in humans. I will use a CRISPR system to establish a novel high-throughput high-content screening platform in differentiated adipocytes. Since most lipid modifying genes have been discovered in non-mammalian model systems such as C. elegans, Drosophila and S. cerevisiae using siRNA approaches, I aim to develop a novel superior CRISPR screening platform in mammalian system. I will take advantage of inducible CRISPRi mouse our lab has developed to perform a single and dual genetic perturbation screen in primary adipocytes. Thus, my proposed research strategy will illuminate not only new genes but importantly genetic interactions and pathways responsible for formation and breakdown of lipid droplets and provide new targets for anti-obesity therapy.

项目摘要肥胖和相关代谢紊乱是影响全世界数百万人的重大医疗保健危机并且没有长期治疗。我们目前关于细胞如何调节新陈代谢的大部分知识是使用单基因驱动的假设获得的。然而，肥胖像许多其他疾病一样是多基因的疾病，其中多个基因相互作用，导致表型，这使得不可能建立致病等位基因。如果我们要在此期间在治疗肥胖和代谢疾病方面取得任何进展本世纪，我们需要大胆、有创意的想法来解决细胞途径，而不是单独的基因尖端高通量方法的优势。截至 2016 年，全球估计有超过 19 亿成年人超重，6.5 亿成年人肥胖。肥胖的典型特征是脂肪组织的病理发育和分化，这是一种体内重要的能量储存器官。脂肪组织将能量储存在脂滴中——直到最近才被人们认识到具有复杂和动态功能的细胞器。脂滴的大小可以发生剧烈变化对热量摄入或新陈代谢变化的反应。由于目前 FDA 批准的抗肥胖药物作用于食欲或代谢调节而不是直接影响脂肪组织脂质，我们需要确定途径负责脂滴的生物合成和分解，以改善当前的治疗方法。令人惊讶的是，很少已知影响人类脂滴的途径。我将利用CRISPR系统建立一个新型的高通量高内涵的差异化筛选平台脂肪细胞。由于大多数脂质修饰基因是在非哺乳动物模型系统中发现的，例如使用 siRNA 方法对秀丽隐杆线虫、果蝇和酿酒酵母进行研究，我的目标是开发一种新型的卓越 CRISPR 哺乳动物系统中的筛选平台。我将利用我们实验室拥有的诱导型 CRISPRi 小鼠开发用于在原代脂肪细胞中进行单基因扰动和双基因扰动筛选。因此，我提出的研究策略不仅将阐明新基因，而且重要的是遗传相互作用和途径负责脂滴的形成和分解，并为抗肥胖治疗提供新的靶点。