Regulation of Liver Metabolism by lncRNAs

项目摘要 目前的研究计划是研究一种新的胰岛素依赖分子Gm11967的生理功能 在我目前的K01奖中发现了长的非编码RNA(LncRNA)。尽管ncRNAs占 大多数转录组，对其生物学意义和功能靶点知之甚少。 LncRNAs是一大类多种多样的RNA转录本，其长度超过200个核苷酸，而不是 编码蛋白质。到目前为止，很少有lncRNA被详细描述；然而，最近的工作已经 提示了lncRNA在糖尿病和肥胖症等代谢紊乱中的潜在作用。因此，我们重新- 分析了在我的K01奖项期间产生的全基因组数据集，以询问肝脏胰岛素的作用 对lncRNA表达的信号转导以识别新的、代谢调节的lncRNA。我们确定Gm11967是 在体内诱导肝脏中最显著的lncRNA之一。利用小鼠遗传学，我们证明了Gm11967 是Akt-Foxo1轴的下游，Akt-Foxo1轴是参与肝脏葡萄糖和脂肪代谢的中央信号节点。 Gm11967与葡萄糖激酶共享一个转录起始点，后者是一种有效的胰岛素调节基因，是 我现在的K01。考虑到它对进食的强烈激活，对胰岛素作用的依赖，以及密切的联系 通过葡萄糖激酶的表达，我们假设Gm11967在肝脏的调节中起着中心作用。 糖脂代谢。我们将在体内进行功能获得和丧失的实验，以阐明 Gm11967在肝细胞中的生物学功能此外，我们还将通过RNA进行染色质的分离 纯化(CHIRP)，然后进行质谱分析和/或深度测序，以鉴定功能蛋白质组和 Gm11967的基因组靶点。根据我的培训和在我的K01课程期间获得的专业知识，我已经 完成这项提议所需的独立性和技术技能。重要的是，这些数据将建立 在我的K01的坚实基础上，并将实现导致未来R01的新的调查路线 申请。

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.