Identification and Characterization of Novel Metabolic Regulators in Mouse and Human Liver

项目总结/摘要 长链非编码RNA（lncRNA）是一类新兴的和快速增长的功能基因组元件， 和一些已被证明可以调节基本的生物过程，但它们的影响范围 在代谢紊乱中的作用尚不清楚。识别和表征代谢相关的lncRNA将是 这对更好地理解肥胖和其他代谢疾病的病理生理学至关重要。 候选人渴望通过研究新的代谢调节剂来促进这种理解， 发展相关研究技术的额外经验，并获得必要的职业 发展培训，以过渡到独立主要研究者的角色。 扩大候选人最近的全基因组筛选，确定了140个lncRNA和239个mRNA敏感 对小鼠肝脏代谢条件的影响，拟议的研究将研究一种新的lncRNA，Gm 15441， 在调节甘油三酯（TG）和胆固醇代谢（Aim 1）和一个新的蛋白质编码基因Rab 30， 调节小鼠肝脏中的自噬、游离脂肪酸（FFA）氧化和TG分泌（目的2）。机制 Gm 15441调节TG和胆固醇代谢的机制将通过探索其对TG和胆固醇代谢的顺式调节来确定。 重叠代谢相关蛋白质编码基因。在初步实验中，Rab 30的敲除 影响了禁食小鼠肝脏的自噬和脂质代谢，因此Rab 30在膜 贩运将被确定。此外，在生物信息学领域建立的全面的生物信息学分析管道 最近的全基因组筛选将应用于人类肝脏样本，以确定新的lncRNA代谢调节因子 在人类肝脏中（目标3）。建立了该管道预测人类基因功能的功效 在初步实验中，通过测试已知的代谢基因，如G6 PC（葡萄糖-6-磷酸酶催化 亚基），这是正确预测的功能，在胚胎发育。 K22奖将为这位候选人提供机会，发展CRISPR，共聚焦显微镜，细胞 生物学和生物信息学。候选人将发展这些技能，同时发展追求 翻译意义的研究。例如，gm 15441位于与 人类1 q21 -23基因座与2型糖尿病和家族性混合型高脂血症相关。候选 已经为她的职业发展制定了一个全面的计划，已经确定了课程，将赋予 作为一名独立调查员，成功的必要技能，并获得了一个杰出的导师团队 以及在她的职业转型期间为她提供支持的顾问。成功完成所述工作 在这个提议中，可以提供对肝脏和全身代谢的调节网络的重要见解， 并将为候选人提供实现研究独立的能力。

Project Summary/Abstract The long non-coding RNAs (lncRNAs) are an emerging and rapidly-growing class of functional genomic elements, and a number have been shown to regulate fundamental biological processes, but the scope of their influence in metabolic disorders remains unknown. Identifying and characterizing metabolically-relevant lncRNAs will be crucial to obtaining a better understanding the pathophysiology of obesity and other diseases of metabolism. The candidate aspires to contribute to this understanding by studying novel metabolic regulators, while developing additional experience in relevant research techniques and receiving the necessary career development training to transition to a role as an independent principal investigator. Expanding on the candidate’s recent genome-wide screen that identified 140 lncRNAs and 239 mRNAs sensitive to metabolic conditions in mouse liver, the proposed research will study the roles of one novel lncRNA, Gm15441, in regulating triglyceride (TG) and cholesterol metabolism (Aim 1) and one novel protein-coding gene, Rab30, in regulating autophagy, free fatty acid (FFA) oxidation, and TG secretion in mouse livers (Aim 2). The mechanism by which Gm15441 regulates TG and cholesterol metabolism will be defined by exploring its cis-regulation of the overlapping metabolically-relevant protein-coding gene. In preliminary experiments, knockdown of Rab30 affected autophagy and lipid metabolism in fasting mouse livers, therefore the role of Rab30 in membrane trafficking will be determined. Additionally, the comprehensive bioinformatics analysis pipeline established in the recent genome-wide screen will be applied to human liver samples, to identify novel lncRNA metabolic regulators in human liver (Aim 3). The efficacy of this pipeline for predicting the function of human genes was established in preliminary experiments by testing known metabolic genes, such as G6PC (glucose-6-phosphatase catalytic subunit), which was correctly predicted to function in gluconeogenesis. A K22 award will offer this candidate the opportunity to develop new skills in CRISPR, confocal microscopy, cell biology, and bioinformatics. The candidate will develop these skills while developing experience in pursuing research of translational significance. Gm15441, for instance, is localized in a region that is syntenic with the human 1q21-23 locus, implicated in type 2 diabetes mellitus and familial combined hyperlipidemia. The candidate has developed a comprehensive plan for the development of her career, has identified coursework that will confer necessary skills for success as an independent investigator, and has secured a team of distinguished mentors and advisors who will support her throughout her career transition. Successful completion of the work described in this proposal could provide critical insight into the regulatory networks of hepatic and systemic metabolism, and will afford the candidate the ability to achieve research independence.