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）使能技术和特定的挑战主题，06-GM-105：小RNA。近60%的美国成年人肥胖或超重。清楚地了解基因调控的基本机制，以应对能源供应的变化是临床，制药和社会的重要性。本研究的长期目标是验证以下假设：下丘脑miRNA水平的调节随着能量可用性的变化而发生，靶向控制体重的特定基因的表达。每个实验室的综合专业知识加强了这种多PI提案。Good博士的实验室在研究下丘脑基因对能量可用性变化的转录控制方面有超过10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享誉世界。Helm博士的实验室使用蛋白质组学和代谢组学方法来研究细胞过程，如静止。这三个PI将共同使用miRNA和mRNA微阵列，蛋白质组学和生物信息学来询问有关miRNA在体重调节中的作用的具体和基本问题，包括：（i）miRNA种类是否响应于能量可用性的变化而受到差异调节（ii）这些miRNA种类是否靶向调节体重控制的特定下丘脑基因。将通过三管齐下的生物信息学方法鉴定共调节的miRNA、mRNA和蛋白质，以检查鉴定的miRNA种类的推定靶标的表达水平，并将这些与靶基因的蛋白质和mRNA水平之间的可能不一致进行比较。这是一种创新的方法，尚未对整个动物组织进行过。这些研究通过（1）建议研究miRNA在肥胖中的作用的科学挑战-我们知识中的空白，特别是因为它与下丘脑基因调控有关;（2）提出一项旨在通过这些为期两年的刺激资金使科学界受益的研究，通过生成至少3个新的数据集，供我们和科学界的其他人在未来的研究中使用;（3）提出一个研究设计，其研究方法将受益于大量资金的涌入，以显著的方式快速推进miRNA研究领域。随着肥胖症达到创纪录的比例，发现新的基因调控机制将导致重要的新的临床，制药和基础研究方向和应用。 公共卫生相关性：拟议的实验将表征一种潜在的新方法，在食物摄入，食物剥夺或肥胖个体与高水平的循环瘦素的下丘脑基因调控。了解大脑中的这些分子途径，并识别在食物供应变化期间差异调节的基因和蛋白质，应该为肥胖和超重个体找到新的治疗选择，并提供有关我们大脑的基本机制的新信息用于控制体重。

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.