Small RNAs: Regulation and Regulatory Activity in Body Weight Control

描述(由申请人提供)：本申请涉及广泛的挑战领域(06)使能技术和具体的挑战主题06-GM-105：小RNA。近60%的美国成年人口要么肥胖，要么超重。清楚地了解基因调控响应能量供应变化的基本机制具有临床、药物和社会重要性。这项研究的长期目标是检验这一假设，即下丘脑miRNA水平的调节发生在以控制体重的特定基因的表达为目标的能量可获得性变化的过程中。通过每个实验室的综合专业知识，这一多重PI建议得到加强。古德博士的实验室在研究下丘脑基因转录控制以应对能量供应变化方面拥有10多年的经验。詹森博士因其在微阵列质量和基因表达分析评估方面的工作而闻名于世。赫尔姆博士的实验室使用蛋白质组学和新陈代谢组学的方法来研究细胞过程，如静止。这三个PI将一起使用miRNA和mRNA微阵列、蛋白质组学和生物信息学来询问关于miRNA在体重调节中的作用的具体和基本的问题，包括：(I)miRNA物种是否因能量可获得性的变化而进行差异调控(Ii)这些miRNA物种是否针对调节体重控制的特定下丘脑基因。共同调控的miRNA、mRNAs和蛋白质将通过三管齐下的生物信息学方法进行鉴定，以检查所识别的miRNA物种的假定靶标的表达水平，并将这些表达水平与靶基因的蛋白质和信使核糖核酸水平之间可能的不一致性进行比较。这是一种创新的方法，还没有对整个动物组织进行过研究。这些研究通过(1)建议研究miRNA在肥胖中的作用这一科学挑战--这是我们知识中的一个空白，特别是与下丘脑基因调控有关的知识；(2)提出一项旨在通过这些为期两年的刺激基金造福科学界的研究，为我们和科学界的其他人产生至少3个新的数据集，用于未来的研究；以及(3)提出一项研究设计，其研究方法将受益于资金的涌入，以显著促进miRNA研究领域的快速发展。随着肥胖比例的创纪录，新的基因调控机制的发现将导致重要的新的临床、药物和基础研究方向和应用。 与公共健康相关：拟议的实验将表征一种潜在的新方法，用于在食物摄取、食物剥夺或循环瘦素水平较高的肥胖者中调节下丘脑基因。了解大脑中的这些分子途径，并识别在食物可获得性变化过程中差异调节的基因和蛋白质，应该会确定肥胖和超重个体的新治疗选择，并提供关于我们大脑用来控制体重的基本机制的新信息。

DESCRIPTION (provided by applicant): This application addresses the broad Challenge Area (06) Enabling Technologies and the specific Challenge Topic, 06-GM-105: Small RNAs. Nearly 60% of the US adult population is either obese or overweight. Clearly understanding the fundamental mechanisms of gene regulation in response to changes in energy availability is of clinical, pharmaceutical, and societal importance. The long-term objective of this research is to test the hypothesis that regulation of hypothalamic miRNA levels occurs with changes in energy availability targeting the expression of specific genes that control body weight. This multiple PI proposal is strengthened by the combined expertise of each individual laboratory. Dr. Good's lab has more than 10 years experience in studying the transcriptional control of hypothalamic genes in response to changes in energy availability. Dr. Jensen is world-renowned for his work on microarray quality and assessment of gene expression analyses. Dr. Helm's lab uses proteomic and metabolomic approaches to study cellular processes such as quiescence. Together, the three PIs will use miRNA and mRNA microarrays, proteomics and bioinformatics to ask specific and fundamental questions about the role of miRNA in body weight regulation including: (i) whether miRNA species are differentially-regulated in response to changes in energy availability (ii) if these miRNA species target specific hypothalamic genes that regulate body weight control. The co-regulated miRNA, mRNAs and proteins will be identified through a three-pronged bioinformatics approach to examine the expression levels of the putative targets of the identified miRNA species and compare these with possible discordance between the protein and mRNA levels for the target genes. This is an innovative method that has not yet been done for whole animal tissues. These studies specifically address the intent of the NIH Challenge Grants by (1) proposing to study the scientific challenge of the role of miRNA in obesity-a gap in our knowledge, especially as it relates to hypothalamic gene regulation; (2) proposing a study that is designed to benefit the scientific community through these 2-year stimulus funds, by generating at least 3 new datasets for us and others in the scientific community to use in future studies; and (3) proposing a study design with research methods that will benefit from an influx of funds to quickly advance the area of miRNA research in significant ways. With obesity at record proportions, the discovery of new mechanisms of gene regulation will lead to important new clinical, pharmaceutical and basic research directions and applications. PUBLIC HEALTH RELEVANCE: The proposed experiments will characterize a potentially new method for hypothalamic gene regulation during food intake, food deprivation or in obese individuals with high levels of circulating leptin. An understanding of these molecular pathways in the brain, and identification of the genes and proteins that are differentially regulated during changes in food availability should identify new therapeutic options for obese and overweight individuals, as well as give new information about the basic mechanisms that our brains use to control body weight.