Alternative polyadenylation as a genetic regulatory mechanism to bridge genome to phenome in the nervous system

替代多腺苷酸化作为连接神经系统基因组和表型的遗传调控机制

• 批准号: 10541679
• 负责人: Julianna Nicole Brutman
• 金额: $ 3.68万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10541679
• 关键词: Anorexia; Anorexia Nervosa; Appetite Alteration; Appetite Disorder; Appetite Regulation; Appetitive Behavior; Behavioral; Bioinformatics; Biology; Body Weight; Cancer Patient; Cannabis sativa plant; Cell Culture Techniques; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Computer Models; Data; Dependovirus; Desire for food; Development; Diabetes Mellitus; Disease; Eating; Eating Disorders; Ensure; Exhibits; Exposure to; Feeding Patterns; Feeding behaviors; Food; Genes; Genetic; Genome; Goals; Health; Heart Diseases; Hyperphagia; Hypothalamic structure; Institution; Investigation; Knock-out; Knockout Mice; Knowledge; Lead; Learning; Link; Malignant Neoplasms; Maps; Mediating; Mental disorders; Messenger RNA; Metabolic; Molds; Molecular; Mus; Nervous system structure; Neuraxis; Neurobiology; Neurons; Neurosciences; Neurosciences Research; Nuclear Export; Obesity; Operative Surgical Procedures; Palate; Pathologic; Pathway interactions; Pattern; Peptide Hydrolases; Pharmaceutical Preparations; Phase; Phenotype; Polyadenylation; Positioning Attribute; Process; Protein Array; Proteins; Proteomics; Psychopathology; Publishing; RNA; RNA Processing; RNA Splicing; RNA Stability; Rattus; Regulation; Research; Research Institute; Research Personnel; Research Project Grants; Research Training; Rodent; Site; Structure of nucleus infundibularis hypothalami; Synaptic plasticity; Testing; Tissue Inhibitor of Metalloproteinases; Tissue-Specific Gene Expression; Training; Training Support; Transcript; United States; Universities; Washington; Weight Gain; Work; base; behavior influence; behavioral phenotyping; cancer cachexia; career; comorbidity; design; design verification; diet-induced obesity; energy balance; experimental study; feeding; gene product; innovation; mRNA Expression; metabolic phenotype; meter; mortality; mouse model; multiple omics; neurogenetics; novel; obesity development; overweight adults; phenome; pre-doctoral; professor; protein expression; single cell sequencing; skills; small hairpin RNA; synaptic function; targeted treatment; vector

PROJECT SUMMARY Anorexia nervosa is the deadliest psychological disorder with an estimated 10% lifetime disease mortality rate, while over 1/3 of all cancer patients will die from disease-based anorexia, not the cancer itself. On the opposite end of the spectrum, over 2/3 of US adults are overweight or obese, and this number, as well as the rates of associated comorbidities such as heart disease, diabetes, and cancer, is only expected to increase in the coming years. Despite the opposite directionality of these eating disorders, dysfunctional eating in obesity and anorexia is mediated by common appetite circuitry in the central nervous system (CNS). Numerous studies have documented a coordinated and complex pattern of changes in multiple gene products in these appetite centers following periods of excessive or inadequate eating. These observations strongly suggest that the behavioral decision to eat excessively or inadequately is likely driven by a multitargeted, maladaptive genetic reprogramming process in CNS appetite centers. Thus, a core question is what global process could coordinate such changes in multiple gene products? My published studies have demonstrated that appetite changes align with changes in alternative polyadenylation (APA) in the hypothalamus. APA is a rapid, activity-dependent RNA processing mechanism that regulates mRNA transcript stability, maturation, and localization. I identified a significant APA pattern change on tissue inhibitor of metalloproteinases 2 (Timp2), a gene previously implicated in the development of an obese phenotype. Thus, I am exploring the hypothesis that Timp2 APA in the arcuate nucleus (ARC) of the hypothalamus meters the development of obesity. My proposed experiments in the F99 phase will show that 1) Timp2 mRNA is necessary for appetite control in the ARC, and that 2) APA regulation of ARC Timp2 is necessary to counteract hyperphagia and obesity. These studies will be the first to functionally link APA regulation to feeding behavior and will serve as the basis of further genome to behavioral phenome studies in my independent career. My Sponsor, Dr. Gary Wayman, and Co-Sponsors, Drs. Suzanne Appleyard and Emily Qualls-Creekmore, are established neuroscientists at Washington State University with expertise in molecular neuroscience (Wayman) and ingestive behavior (Appleyard and Qualls-Creekmore). My proposed Research and Training plan will strengthen my theoretical and technical understanding of neurogenetics. In the pre-doctoral F99 phase, I will learn shRNA and CRISPR/SaCas9 vector design and validation strategies, cell culture techniques, stereotaxic surgeries, and advanced metabolic analyses. In the postdoctoral K00 phase, I will build upon these skills and learn to use genetic mouse models, multi-omics, advanced bioinformatics, and AI computational models to map genome to phenome regulation. Overall, the proposed training will optimally position me to start an independent research career at a leading neuroscience research institute and advance our understanding of RNA regulation as a functional link between the genome and the behavioral phenome.

项目概要 神经性厌食症是最致命的心理疾病，估计终身疾病死亡率为 10% 率，而超过 1/3 的癌症患者将死于疾病性厌食症，而不是癌症本身。上 与此相反，超过 2/3 的美国成年人超重或肥胖，这个数字以及 心脏病、糖尿病和癌症等相关合并症的发病率预计只会在 未来几年。尽管这些饮食失调的方向相反，但肥胖症患者的饮食失调 厌食症是由中枢神经系统（CNS）中常见的食欲回路介导的。大量研究 已经记录了这些食欲中多种基因产物的协调和复杂的变化模式 饮食过量或不足后的中心。这些观察强烈表明 吃得过多或不足的行为决定可能是由多目标、适应不良的遗传因素驱动的 中枢神经系统食欲中心的重编程过程。因此，一个核心问题是全球进程可以协调什么 多个基因产物发生如此变化？我发表的研究表明，食欲的变化是一致的 下丘脑中选择性多聚腺苷酸化（APA）的变化。 APA 是一种快速、活性依赖性的 RNA 调节 mRNA 转录稳定性、成熟和定位的加工机制。我确定了一个 金属蛋白酶组织抑制剂 2 (Timp2)（先前涉及的基因）的 APA 模式发生显着变化 肥胖表型的发展。因此，我正在探索 Timp2 APA 在弓状体中的假设 下丘脑核（ARC）控制肥胖的发展。我在 F99 中提出的实验 阶段将表明 1) Timp2 mRNA 对于 ARC 中的食欲控制是必需的，并且 2) APA 调节 ARC Timp2 对于对抗食欲过盛和肥胖是必需的。这些研究将是第一个从功能上 将 APA 调节与进食行为联系起来，并将作为进一步基因组到行为现象组的基础 在我的独立职业生涯中学习。我的赞助商加里·韦曼博士和共同赞助商博士。苏珊娜·阿普尔亚德 和 Emily Qualls-Creekmore 是华盛顿州立大学的知名神经科学家，在以下领域拥有专业知识： 分子神经科学（Wayman）和摄入行为（Appleyard 和 Qualls-Creekmore）。我的提议 研究和培训计划将加强我对神经遗传学的理论和技术理解。在 博士前F99阶段，我将学习shRNA和CRISPR/SaCas9载体设计和验证策略，细胞 培养技术、立体定位手术和先进的代谢分析。在博士后K00阶段，我 将在这些技能的基础上学习使用遗传小鼠模型、多组学、先进的生物信息学和 人工智能计算模型将基因组映射到表型调控。总体而言，拟议的培训将最佳地 让我能够在领先的神经科学研究所开始独立的研究生涯并取得进步 我们对 RNA 调控的理解是基因组和行为现象之间的功能联系。