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

• 批准号: 10066969
• 负责人: Olga Gulyaeva
• 金额: $ 6.53万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10066969
• 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; Patients; 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; pandemic disease; programs; public health relevance; response; screening; side effect; uptake; whole genome

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 肥胖和相关的代谢紊乱是一个重大的卫生保健危机，影响到全世界数百万人 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知识， 使用单基因驱动假设获得。然而，肥胖症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，有助于表型，这使得不可能建立 致病等位基因如果我们要在这段时间内在治疗肥胖和代谢疾病方面取得任何进展， 世纪，我们需要大胆的，创造性的想法来解决细胞途径，而不是单个基因， 先进的高通量方法的优势。 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。 肥胖症的典型特征是脂肪组织的病理性发育和分化，这是一种 是身体的主要能量储存器官。脂肪组织在脂滴中储存能量-直到最近才被认识到 具有复杂和动态功能的细胞器。脂滴的大小可以急剧变化， 对卡路里摄入量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，而不是直接对脂肪组织脂质，我们需要确定途径 负责脂滴生物发生和分解以改善当前的治疗。令人惊讶的是，很少 已知影响人类脂滴的途径。 我将使用CRISPR系统建立一个新的高通量高内容筛选平台， 脂肪细胞由于大多数脂质修饰基因已经在非哺乳动物模型系统中发现， C. elegans、Drosophila和S.我的目标是开发一种新的上级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一和双重遗传扰动筛选。因此，我建议 研究战略不仅要阐明新基因，而且要阐明重要的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。