Role of intestinal luminal and epigenetic factors to mediate intestinal metabolic remodeling after gastric bypass surgery

肠腔和表观遗传因素在胃绕道手术后介导肠道代谢重塑中的作用

• 批准号: 10373092
• 负责人: Margaret Stefater-Richards
• 金额: $ 16.79万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10373092
• 关键词: ATAC-seq; Address; Affect; Alternative Therapies; Amino Acids; Animals; Area; Biliary; Bioinformatics; Biology; Blood; Caloric Restriction; Childhood; Chromatin; Chronic; Clinical; Clinical Trials; Data; Dependence; Development; Development Plans; Disease remission; Doctor of Philosophy; Duodenum; Elements; Endocrinologist; Energy Metabolism; Environment; Environmental Risk Factor; Epigenetic Process; Exclusion; Exhibits; Exposure to; Fingerprint; Fostering; Gastrectomy; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genus Hippocampus; Goals; Grant; Growth; Health; Homing; Human; Incidence; Intestinal Bypasses; Intestines; K-Series Research Career Programs; Lead; Life Style; Light; Limb structure; Link; Mediating; Mentorship; Metabolic; Methods; Methylation; Mission; Modeling; Modification; Mus; Muscle; Non-Insulin-Dependent Diabetes Mellitus; Nutrient; Nutritional; Obesity; Operative Surgical Procedures; Organoids; Pathway interactions; Patients; Pharmacology; Physiology; Play; Postoperative Period; Pre-Clinical Model; Public Health; Regulation; Research; Role; Route; Techniques; Testing; Therapeutic; Thinness; Tissue Sample; Tissues; United States National Institutes of Health; Youth; bariatric surgery; base; career development; chromatin remodeling; diabetes mellitus therapy; differential expression; effective intervention; effective therapy; energy balance; epigenetic regulation; experience; experimental study; gene regulatory network; improved; in vivo; innovation; insight; intestinal epithelium; metabolic phenotype; new therapeutic target; obesity development; pediatric patients; prospective; response; transcriptome sequencing; transcriptomics

PROJECT SUMMARY The rising incidence of type 2 diabetes (T2D) highlights a growing need to understand mechanisms behind highly-effective therapies such as Roux-en-Y gastric bypass surgery (RYGB) in order to develop better, more widely-applicable treatments. The long-term goal is to understand the role of luminal factors to mediate the effect of intestinal re-routing upon energy metabolism, including T2D. The overall objective of this application is to test for tissue-level epigenetic changes in the intestine after RYGB, as a potential mechanism for Roux limb (RL) remodeling that we have shown to relate to T2D remission in humans. The central hypothesis is that RL metabolic reprogramming is driven by segment-specific epigenetic modifications which are plastic and require continuous luminal stimulation. The rationale for this project is that a greater understanding of intestinal gene expression regulatory networks after RYGB will shed light on ways to mimic RYGB’s effects with less invasive alternative therapies. The central hypothesis will be tested in two specific aims: (1) to test the role of an unexpected luminal nutrient load to influence RL epigenetic and transcriptomic signatures for key genes associated with metabolically-significant RL remodeling; and (2) to test the dependency of RL epigenetic signatures and metabolic substrate utilization upon in vivo factors using intestinal epithelial organoids (IEOs). In aim 1, gene expression (RNA-seq) and chromatin remodeling (ATAC-seq) will be studied in RL versus bypassed intestinal limb (biliopancreatic limb) or versus sleeve gastrectomy, and these methods will also be employed to test the plasticity of RL remodeling during caloric restriction and refeeding. In aim 2, IEOs will be used to ask whether epigenetic (ATAC-seq) and metabolic reprogramming (RNA-seq; Seahorse XF Analysis) change in the absence of in vivo factors. The candidate for this career development award, Dr. Margaret Stefater, is a pediatric endocrinologist with an MD/PhD and expertise in the integrated physiology of energy balance, especially after bariatric surgery. In order to achieve scientific independence for the candidate, a comprehensive career development plan has been developed to gain additional experience in the areas of (1) epigenetics and gene regulatory networks, (2) bioinformatics, and (3) use of intestinal organoids as a model of intestinal biology. This will be accomplished through coursework and seminars, under the mentorship of experts in the fields of bariatric surgery (Nicholas Stylopoulos) and intestinal biology (David Breault). The proposed research in this application is innovative, in the applicant’s opinion, because it represents a substantial departure from the status quo by using state-of-the-art sequencing and bioinformatics techniques to address how RYGB’s core surgical elements (e.g., biliary exclusion, accelerated nutrient delivery) influence epigenetic regulation of gene expression. Ultimately, this project will shed light on mechanisms linking luminal factors to known intestinal remodeling after RYGB, thus highlighting potential novel drug targets for the development of innovative T2D therapies, and paving the way for the applicant’s first R01 submission.

项目摘要 2型糖尿病（T2 D）发病率的上升凸显了对了解其背后机制的日益增长的需求。 高效的治疗方法，如Roux-en-Y胃旁路手术（RYGB），以开发更好，更 广泛适用的治疗。长期的目标是了解管腔因子在介导细胞凋亡中的作用。 肠道改道对能量代谢的影响，包括T2 D。本申请的总体目标是 检测RYGB后肠道组织水平的表观遗传变化，作为Roux肢体的潜在机制 (RL)我们已经证明与人类T2 D缓解有关的重塑。核心假设是RL 代谢重编程是由片段特异性表观遗传修饰驱动的，这些表观遗传修饰是可塑的， 连续的腔刺激。这个项目的基本原理是， RYGB后的表达调控网络将揭示如何模拟RYGB的影响， 替代疗法中心假设将在两个特定的目标进行测试：（1）测试的作用， 意想不到的管腔营养负荷影响RL表观遗传和转录组特征的关键基因 与代谢显着的RL重塑相关;和（2）测试RL表观遗传的依赖性 使用肠上皮类器官（IEO）的体内因子的特征和代谢底物利用。 在目标1中，将在RL与对照组中研究基因表达（RNA-seq）和染色质重塑（ATAC-seq）。 旁路肠分支（胆胰分支）或相对于袖状胃切除术，这些方法也将 用于测试在热量限制和再喂养期间RL重塑的可塑性。在目标2中，IEO将 用于询问表观遗传（ATAC-seq）和代谢重编程（RNA-seq;海马XF分析） 在缺乏体内因素的情况下发生变化。这个职业发展奖的候选人玛格丽特博士 Stefater是一名儿科内分泌学家，拥有医学博士/博士学位，擅长能量综合生理学 平衡，尤其是减肥手术后。为了实现候选人的科学独立性， 制定了全面的职业发展计划，以获得以下领域的额外经验：（1） 表观遗传学和基因调控网络，（2）生物信息学，和（3）使用肠道类器官作为模型， 肠道生物学这将在以下人员的指导下通过课程和研讨会来实现 减肥手术（Nicholas Stylopoulos）和肠道生物学（大卫Breault）领域的专家。的 在申请人看来，本申请中提出的研究是创新的，因为它代表了 通过使用最先进的测序和生物信息学技术， 解决RYGB的核心手术要素（例如，胆汁排除，加速营养输送）影响 基因表达的表观遗传调控。最终，这个项目将揭示连接管腔的机制， RYGB后已知的肠重塑因素，从而突出了潜在的新药物靶点， 开发创新的T2 D疗法，并为申请人的第一个R 01申请铺平道路。