Regulation of Liver Metabolism by lncRNAs

lncRNA 对肝脏代谢的调节

• 批准号: 9975166
• 负责人: Paul Michael Titchenell
• 金额: $ 7.73万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-09 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9975166
• 关键词: Accounting; Affect; Affinity; Biological; Biological Assay; Biological Process; Cell Line; Chromatin; Code; Complex; DNA; Data; Data Set; Dependence; Diabetes Mellitus; Enhancers; FOXO1A gene; Foundations; Funding; Future; Gene Expression; Genes; Genetic Transcription; Genetic Translation; Genome; Genomics; Glucokinase; Glucose; Goals; Hepatic; Hepatocyte; Homeostasis; Hyperinsulinism; Individual; Insulin; Intake; Investigation; Length; Lipids; Liver; Mass Spectrum Analysis; Measures; Mediating; Mentored Research Scientist Development Award; Mentorship; Metabolic; Metabolic Diseases; Metabolic Pathway; Metabolism; Mus; Myocardium; National Institute of Diabetes and Digestive and Kidney Diseases; Nuclear; Nucleotides; Nutrient; Obesity; Oligonucleotides; Open Reading Frames; Physiological; Play; Precipitation; Protein-Serine-Threonine Kinases; Proteins; Proteome; Proteomics; Publications; RNA; RNA Binding; RNA Splicing; RNA purification; Regulation; Research Proposals; Role; Running; Signal Transduction; Sonication; Techniques; Testing; Tissue Sample; Tissues; Training; Transcript; Transcription Initiation Site; Untranslated RNA; Work; base; chromatin remodeling; crosslink; deep sequencing; design; experience; experimental study; feeding; gain of function; genome-wide analysis; global run on sequencing; glucose metabolism; glucose tolerance; hormonal signals; in vivo; insulin regulation; insulin signaling; insulin tolerance; knock-down; lipid metabolism; liver metabolism; loss of function; mRNA Precursor; mouse genetics; novel; overexpression; response; transcriptome

Project Summary The current research proposal is to investigate the physiological function of Gm11967, a novel insulin-dependent long non-coding RNA (lncRNA) identified during my current K01 award. Despite ncRNAs accounting for a majority of the transcriptome, very little is known about their biological significance and functional targets. lncRNAs are a large and diverse class of RNA transcripts with a length of more than 200 nucleotides that do not encode proteins. To date, very few lncRNAs have been characterized in detail; however, recent work has suggested a potential role for lncRNA in metabolic disorders such as diabetes and obesity. Therefore, we re- analyzed genome-wide data sets generated during my K01 award to interrogate the role of hepatic insulin signaling on lncRNA expression to identify novel, metabolically regulated lncRNAs. We identified Gm11967 as one of the most significantly induced lncRNA in liver in vivo. Using mouse genetics, we demonstrated Gm11967 is downstream of the Akt-Foxo1 axis, a central signaling node involved in hepatic glucose and lipid metabolism. Gm11967 shares a transcription start site with glucokinase, a potent insulin-regulated gene that is the focus of my current K01. Given its robust activation with feeding, dependency on insulin action, and close association with glucokinase expression, we hypothesized that Gm11967 plays a central role in the regulation of hepatic glucose and lipid metabolism. We will perform gain and loss of function experiments in vivo to elucidate the biological function of Gm11967 in hepatocytes. In addition, we will perform chromatin isolation by RNA purification (ChIRP) followed by mass spectrometry and/or deep sequencing to identify functional proteomic and genomic targets of Gm11967. Based on my training and expertise obtained during the course of my K01, I have the independence and technical skillsets necessary to complete this proposal. Importantly, these data will build upon the strong foundation of my K01 and will enable new lines of investigation that will lead to a future R01 application.

项目摘要 目前的研究计划是研究一种新的胰岛素依赖性胰岛素受体Gm 11967的生理功能。 长链非编码RNA（lncRNA）在我目前的K 01奖期间鉴定。尽管ncRNA占 对于大多数转录组，对其生物学意义和功能靶点知之甚少。 lncRNA是一种长度超过200个核苷酸的大而多样的RNA转录物， 编码蛋白质。迄今为止，很少有lncRNA被详细表征;然而，最近的工作已经 提示lncRNA在代谢紊乱如糖尿病和肥胖症中的潜在作用。因此，我们重新- 我分析了在我的K 01奖期间产生的全基因组数据集，以询问肝胰岛素的作用， 信号转导对lncRNA表达的影响，以鉴定新的代谢调节的lncRNA。我们将GM 11967鉴定为 在体内肝脏中最显著诱导的lncRNA之一。利用小鼠遗传学，我们证明了Gm 11967 位于Akt-Foxo 1轴的下游，Akt-Foxo 1轴是参与肝脏葡萄糖和脂质代谢的中心信号节点。 GM 11967与葡萄糖激酶共享一个转录起始位点，葡萄糖激酶是一种有效的胰岛素调节基因， 现在的K 01鉴于其与进食的强烈激活，对胰岛素作用的依赖性，以及与胰岛素的密切相关性， 在葡萄糖激酶表达的情况下，我们假设Gm 11967在肝细胞凋亡的调节中起核心作用。 葡萄糖和脂质代谢。我们将在体内进行功能获得和丧失实验，以阐明 Gm 11967在肝细胞中的生物学功能。此外，我们将通过RNA进行染色质分离， 纯化（ChIRP），然后进行质谱和/或深度测序，以鉴定功能性蛋白质组和 GM 11967的基因组靶点。根据我在K 01课程中获得的培训和专业知识，我 独立性和完成此提案所需的技术技能。重要的是，这些数据将建立 在我的K 01的坚实基础上，并将使新的调查路线，将导致未来的R 01 应用程序.