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

小鼠和人类肝脏中新型代谢调节剂的鉴定和表征

• 批准号: 9762204
• 负责人: Ling Yang
• 金额: $ 24.38万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9762204
• 关键词: 1q21; AGFG1 gene; Affect; Autophagocytosis; Autophagosome; Binding; Binding Proteins; Bioinformatics; Biological Process; CREB1 gene; Cancer Patient; Cancerous; Career Mobility; Catalytic Domain; Cellular biology; Chinese People; Cholesterol; Cholesterol Homeostasis; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Confocal Microscopy; Data; Data Set; Development Plans; Diabetes Mellitus; Disease; Elements; Epidemic; Familial Combined Hyperlipidemia; Fasting; Fatty Liver; Functional disorder; Genes; Genetic Transcription; Glucagon; Gluconeogenesis; Golgi Apparatus; Health; Hepatic; Hepatocyte; Human; In Vitro; K22 Award; Ketone Bodies; Lipids; Liver; Liver diseases; Malignant Neoplasms; Measures; Mediating; Membrane; Mentors; Messenger RNA; Metabolic; Metabolic Diseases; Metabolic Pathway; Metabolic hormone; Metabolism; Mitochondria; Mus; Non-Insulin-Dependent Diabetes Mellitus; Nonesterified Fatty Acids; Normal tissue morphology; Nutrient; Obese Mice; Obesity; Obesity associated disease; Ontology; Patients; Plasma; Primary carcinoma of the liver cells; Principal Investigator; Protein Family; Proteins; Quantitative Reverse Transcriptase PCR; RNA; Regulation; Research; Research Personnel; Research Technics; Role; Sampling; Secure; Starvation; TXN gene; TXNIP gene; Testing; Therapeutic; Tissues; Training; Transcription Process; Translational Research; Triglyceride Metabolism; Triglycerides; Untranslated RNA; Work; analysis pipeline; career development; chromatin immunoprecipitation; differential expression; experience; experimental study; fatty acid oxidation; functional genomics; genome wide screen; genome-wide; glucose metabolism; glucose-6-phosphatase; in vivo; insight; knock-down; lipid metabolism; liver biopsy; member; new therapeutic target; novel; novel strategies; overexpression; promoter; protein biomarkers; skills; success; trafficking; transcription factor; transcriptome; transcriptome sequencing

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.

项目摘要/摘要 长的非编码RNA（LNCRNA）是一种新兴且快速增长的功能基因组元素， 并且已经证明有许多可以调节基本生物学过程，但其影响的范围 在代谢障碍中仍然未知。识别和表征与代谢相关的LNCRNA将是 对于更好地了解肥胖症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型代谢调节剂来为这种理解做出贡献，而 开发有关相关研究技术并获得必要职业的额外经验 发展培训以过渡到独立首席研究员的角色。 扩展候选人最近的全基因组屏幕，该屏幕确定了140个LNCRNA和239个mRNA敏感 对于小鼠肝脏中的代谢条件，拟议的研究将研究一种新型lncrna，GM15441， 在调节甘油三酸酯（TG）和胆固醇代谢（AIM 1）和一种新型蛋白质编码基因Rab30中 调节自噬，游离脂肪酸（FFA）氧化和小鼠生命中的TG分泌（AIM 2）。机制 GM15441通过探索其顺式调节来调节TG和胆固醇代谢 与代谢相关的蛋白质编码基因重叠。在初步实验中，敲除rab30 在禁食小鼠生活中影响自噬和脂质代谢，因此Rab30在膜中的作用 将确定贩运。此外，在 最近全基因组筛查将应用于人肝样品，以鉴定新的LNCRNA代谢调节剂 在人类肝脏中（AIM 3）。该管道预测人类基因功能的效率已建立 在初步实验中，通过测试已知的代谢基因，例如G6PC（葡萄糖-6-磷酸酶催化 亚基），正确预测在糖异生中起作用。 K22奖将为该候选人提供开发CRISPR，共聚焦显微镜，细胞的新技能的机会 生物学和生物信息学。候选人将在发展追求经验的同时发展这些技能 转化意义的研究。例如，GM15441位于与 人类1q21-23基因座，涉及2型糖尿病和家庭组合高脂血症。候选人 为她的职业发展制定了一项全面的计划，已经确定了课程工作 作为独立调查员成功的必要技能，并获得了杰出导师团队 以及将在整个职业生涯过渡中支持她的顾问。成功完成所描述的工作 在这一建议中，可以为肝和系统性代谢的监管网络提供批判性见解， 并将为候选人提供实现研究独立性的能力。