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.

项目摘要 神经性厌食症是最致命的心理障碍，估计有10％的终身疾病死亡率 比率，而超过1/3的癌症患者将死于基于疾病的厌食症，而不是癌症本身。在 频谱的相对端，超过2/3的美国成年人超重或肥胖，这个数字以及 相关合并症（例如心脏病，糖尿病和癌症）的比率仅有望增加 未来几年。尽管这些饮食失调的方向性相反，但肥胖症的功能失调 厌食症是由中枢神经系统（CNS）中的共同食欲电路介导的。许多研究 已经记录了这些食欲中多个基因产物变化的协调且复杂的模式 过度或不充分饮食之后的中心。这些观察结果强烈表明 过度或不充分饮食的行为决定可能是由多核心，适应不良的遗传驱动的 CNS食欲中心的重编程过程。因此，一个核心问题是全球过程可以协调 多种基因产品的这种变化？我发表的研究表明，食欲变化对齐 下丘脑中替代聚腺苷酸化（APA）的变化。 APA是一种快速，活性依赖的RNA 调节mRNA转录稳定性，成熟和定位的处理机制。我确定了一个 金属蛋白酶2（TIMP2）的组织抑制剂上的显着APA模式变化，这是先前与之相关的基因 在肥胖表型的发展中。因此，我正在探索弧形中TIMP2 APA的假设 下丘脑的核（弧）仪表肥胖的发展。我在F99中提出的实验 阶段将表明1）TIMP2 mRNA对于弧中的食欲控制是必需的，而2）APA调节 ARC TIMP2对于抵消肥大和肥胖是必要的。这些研究将是第一个在功能上 将APA调节与喂养行为联系起来，并将作为行为现象的进一步基因组的基础 在我的独立职业中学习。我的赞助商加里·韦曼（Gary Wayman）博士和共同发起人，博士。 Suzanne Appleyard 和艾米丽·奎尔斯·克里克莫尔（Emily Qualls-Creekmore）是华盛顿州立大学的神经科学家 分子神经科学（Wayman）和摄取行为（Appleyard和Qualls-Creekmore）。我提出的 研究和培训计划将增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矢量设计和验证策略，细胞 培养技术，立体定位手术和高级代谢分析。在博士后K00阶段，我 将以这些技能为基础，并学习使用遗传鼠标模型，多词，高级生物信息学和 AI计算模型将基因组映射到现象组调节。总体而言，拟议的培训将最佳 让我在领先的神经科学研究所开始独立研究职业，并提高 我们对RNA调节作为基因组与行为现象之间的功能联系的理解。