Genomic analysis of C. elegans fat regulatory pathways

线虫脂肪调节途径的基因组分析

• 批准号: 8134753
• 负责人: GARY B RUVKUN
• 金额: $ 39.03万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-15 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8134753
• 关键词: Acetyl-CoA C-Acetyltransferase; Adipocytes; Affect; Animal Feed; Animals; Behavior; Behavioral; Biochemical; Biological Assay; Body fat; Caenorhabditis elegans; Cells; Chimeric Proteins; Cloning; Collection; Couples; Cyclic GMP; Defect; Deposition; Disabled Persons; Endocrine; Escherichia coli; Fatty Acids; Fatty acid glycerol esters; Gap Junctions; Gene Fusion; Gene Mutation; Gene Silencing; Generations; Genes; Genetic; Genetic Epistasis; Genetic Screening; Genetic Transcription; Genome; Genomics; Goals; Haploidy; Health; Homeostasis; Homologous Gene; Human; Induced Mutation; Insulin; Intervention; Intestines; Learning; Length; Leptin; Lipids; Mammals; Maps; Mediating; Metabolic; Metabolic Pathway; Metabolism; Mitochondria; Molecular; Molecular Analysis; Monitor; Morphology; Mus; Mutagenesis; Mutation; Nervous system structure; Neurons; Neuropeptides; Neurosecretory Systems; Nutritional status; Obesity; Oleic Acids; Organ; Other Genetics; Output; Pathway Analysis; Pathway interactions; Pattern; Pharmaceutical Preparations; Phenotype; Phylogeny; Physiological; Production; Proteins; RNA Interference; Receptor Signaling; Refractory; Regulation; Regulator Genes; Regulatory Pathway; Reporter; Reporter Genes; Research Personnel; Resistance; Satiation; Screening procedure; Serotonin; Serotonin Production; Signal Pathway; Signal Transduction; Signaling Pathway Gene; Sorting - Cell Movement; Starvation; Stress; Syndrome; Testing; Tissues; Transcriptional Regulation; base; cell type; driving behavior; drug development; feeding; functional genomics; fusion gene; genetic analysis; genome-wide; high throughput analysis; interest; knock-down; lipid metabolism; mutant; novel; oxidation; positional cloning; promoter; relating to nervous system; research study; response; small molecule

DESCRIPTION (provided by applicant): From a combination of genetic analyses and RNAi screening, we have discovered hundreds of gene inactivations and mutations that promise to reveal the neuroendocrine circuit through which C. elegans fat storage set points are determined. Because RNAi does not as potently target neurons, we have also configured classical genetic screens for low fat storage and high fat storage mutations. Some of the 80 mutants in our current collection store extraordinarily high levels of fat under all conditions tested whereas others have defects in the mobilization of fat induced by starvation or drug treatments. Interestingly, many of the mutants have defects in the behavioral outputs normally induced by starvation or satiety. Thus these mutants do not "feel" starved. We propose to molecular identify 5 of these top candidate mutants per year to discern their molecular identity as well as to delineate from their expression pattern which cells mediate the assessment of fat and the behaviors that drive fat storage. We will determine which of these gene inactivations affect fat levels by regulating rates of feeding and which affect gross metabolic levels. We will determine the cellular focus of gene activity for fat storage and whether any of the genes encode proteins that mediate the actual sorting of fats in the storage organs. From a gene array of starved and well fed animals, we have also discovered a number of genes that are induced by starvation or by feeding. GFP fusions to these genes have generated a robust set of reporters of the starved state. We will cross these reporter genes into our mutant collection to assess which mutants induce a starved state and which do not. In addition, we have used these reporter genes already in a classical genetic screen for mutants that fail to induce a starvation marker gene. C. elegans is amenable to large scale genetic and functional genomic screens which is not feasible in mice. Our worm genomics highlights scores of human genes which are homologues of the worm genes we have identified. In some cases, the genes encode proteins that are attractive for the development of drugs. Therefore, identification of fat storage pathway genes in C. elegans provides targets for intervention of human obesity. PUBLIC HEALTH RELEVANCE Our C. elegans gene inactivation analysis has revealed hundreds of genes that regulate of fat storage, many of which have human homologues. The studies of obesity in C. elegans is likely to identify targets for intervention of obesity in human.

描述（由申请人提供）：结合遗传分析和RNAi筛选，我们发现了数百个基因失活和突变，有望揭示秀丽隐杆线虫脂肪储存设定点的神经内分泌回路。由于RNAi不能有效地靶向神经元，我们还配置了低脂肪储存和高脂肪储存突变的经典遗传筛选。我们目前收集的80个突变体中的一些在所有测试条件下都储存了非常高的脂肪水平，而其他突变体在饥饿或药物治疗诱导的脂肪动员方面存在缺陷。有趣的是，许多突变体在通常由饥饿或饱腹感引起的行为输出中存在缺陷。因此，这些突变体不会“感到”饥饿。我们建议每年对5个这些顶级候选突变体进行分子鉴定，以辨别它们的分子特性，并从它们的表达模式中描绘哪些细胞介导脂肪的评估和驱动脂肪储存的行为。我们将通过调节摄食率来确定哪些基因失活会影响脂肪水平，哪些会影响总代谢水平。我们将确定脂肪储存基因活动的细胞焦点，以及是否有任何基因编码介导脂肪在储存器官中实际分类的蛋白质。从饥饿和喂养良好的动物的基因序列中，我们还发现了一些由饥饿或喂养诱导的基因。与这些基因融合的绿色荧光蛋白产生了一组强有力的饥饿状态报告者。我们将把这些报告基因杂交到我们的突变体中，以评估哪些突变体会诱导饥饿状态，哪些不会。此外，我们已经在经典的基因筛选中使用了这些报告基因，用于无法诱导饥饿标记基因的突变体。秀丽隐杆线虫可以进行大规模的遗传和功能基因组筛选，这在小鼠中是不可行的。我们的蠕虫基因组学突出了人类基因的分数，这些基因是我们已经确定的蠕虫基因的同源物。在某些情况下，这些基因编码的蛋白质对药物开发很有吸引力。因此，秀丽隐杆线虫脂肪储存途径基因的鉴定为干预人类肥胖提供了靶点。我们的秀丽隐杆线虫基因失活分析揭示了数百个调节脂肪储存的基因，其中许多具有人类同源物。秀丽隐杆线虫肥胖的研究有可能为人类肥胖的干预找到靶点。