MicroRNA-375 regulation of enteroendocrine cell biology in diet-induced obesity and bariatric surgery

MicroRNA-375对饮食诱导肥胖和减肥手术中肠内分泌细胞生物学的调节

• 批准号: 10627745
• 负责人: Kieran Louise Koch-Laskowski
• 金额: $ 4.42万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10627745
• 关键词: Address; Adult; Affect; Animal Feed; Animal Model; Animals; Bariatrics; Behavior Therapy; Bioinformatics; Biological; Biological Assay; Body Weight; Body Weight decreased; Cell Lineage; Cell Maintenance; Cells; Cellular biology; Child; Chronic; Circulation; Complex; Data; Data Set; Desire for food; Development; Diet; Dietary Intervention; Disease; Endocrine; Enteroendocrine Cell; Epidemic; Gastrectomy; Gene Expression; Genes; Genomics; Greater curvature of stomach; Health; High Fat Diet; Homeostasis; Hormonal; Hormone secretion; Hormones; Human; Immunohistochemistry; Intestines; Islets of Langerhans; Knockout Mice; Knowledge; Measurement; Mediating; Metabolic; Metabolic Diseases; Metabolic dysfunction; MicroRNAs; Molecular; Mus; Obesity; Obesity Epidemic; Operative Surgical Procedures; Pathogenesis; Pathway interactions; Patients; Pharmacotherapy; Phenotype; Physiological; Physiology; Play; Prevalence; Procedures; Regulation; Resolution; Role; Sampling; Satiation; Secretory Cell; Shapes; Signal Pathway; Signal Transduction; Small Intestines; Small RNA; Stimulus; Stomach; Techniques; Testing; Untranslated RNA; Villus; Weight maintenance regimen; Work; bariatric surgery; blood glucose regulation; candidate identification; cell type; combat; comorbidity; detection of nutrient; diet-induced obesity; dietary; effective therapy; energy balance; environmental change; feeding; gastrointestinal epithelium; global health; glucose tolerance; hormonal signals; improved; in vivo; in vivo Model; interdisciplinary approach; interest; intestinal crypt; intestinal epithelium; intestinal villi; knockout animal; negative affect; new therapeutic target; obesity treatment; obesogenic; pharmacologic; posttranscriptional; preventable death; restoration; single-cell RNA sequencing; stem cells; therapeutic target; transcriptome sequencing; transcriptomics

PROJECT SUMMARY / ABSTRACT Enteroendocrine cells (EECs) coordinate a wide variety of signaling networks to maintain metabolic homeostasis. As a rare secretory cell lineage of the gut epithelium, EECs sense and respond to luminal stimuli by releasing a diverse array of hormones that control nutrient sensing, appetite, glycemic regulation, and energy balance. Diet- induced obesity and bariatric surgery have been associated with the dysregulation and restoration of these hormonal pathways, respectively. Moreover, a growing number of pharmacological strategies have emerged that target key EEC signaling pathways to treat metabolic disease. However, despite these advances the molecular mechanisms regulating EEC biology remain incompletely defined. To address this knowledge gap, this proposal aims to determine the role of an EEC-enriched microRNA (miRNA), miR-375, in regulating the effects of dietary and surgical interventions on EEC biology. MiRNAs are short, non-coding RNA molecules that respond to changing environmental contexts and modulate gene expression at the post-transcriptional level. As such, miRNAs are critical regulators of a myriad of biological pathways, including intestinal epithelial development and function. Our lab has previously demonstrated that miR-375 is highly enriched both in intestinal stem cells (ISCs) and along the EEC lineage, and its expression is dramatically reduced by chronic high-fat diet. In addition, our preliminary data demonstrate significant rescue of miR-375 expression in ISCs following bariatric surgery, coinciding with increases in EEC abundance and circulating gut hormone levels. Therefore, I hypothesize that miR-375 exerts context-specific effects on EEC biology during the pathogenesis and amelioration of diet-induced obesity. The proposed studies will test this hypothesis through an interdisciplinary approach using our lab’s established colony of miR-375 knockout (375-KO) mice together with cutting-edge genomic and bioinformatic techniques. In Aim 1, I will assess how the loss of miR-375 exerts diet-specific effects on the distribution of different EEC subtypes by performing high-resolution single-cell RNA-sequencing (scRNA-seq) of small intestinal crypts and villi from wildtype (WT) and 375-KO animals fed either a chronic chow or high-fat diet. In Aim 2, I will determine how miR-375 contributes to surgically-induced EEC adaptations and metabolic improvements through scRNA-seq analyses of crypt and villus samples from diet-induced obese WT and 375- KO mice following bariatric surgery or a control procedure. With these single-cell datasets, I will bioinformatically determine context-dependent changes in overall EEC abundance, subtype distribution (correlated with circulating gut hormone levels), and gene expression (including identification of candidate miR-375 targets). I will also validate these molecular findings in vivo through immunohistochemical assays and metabolic parameters such as body weight and glucose tolerance. Altogether, these findings will further our understanding of EEC regulation and may provide novel therapeutic targets for the treatment of obesity and its comorbidities.

项目概要/摘要 肠内分泌细胞（EEC）协调多种信号网络以维持代谢稳态。 作为肠道上皮的一种罕见的分泌细胞谱系，EEC 通过释放 控制营养感应、食欲、血糖调节和能量平衡的多种激素。 诱导性肥胖和减肥手术与这些功能失调和恢复有关 此外，越来越多的药理学策略已经出现。 靶向关键的 EEC 信号通路来治疗代谢疾病 然而，尽管有这些进展，但分子生物学仍处于起步阶段。 调节 EEC 生物学的机制仍未完全确定。为了解决这一知识差距，本提案提出。 旨在确定富含 EEC 的 microRNA (miRNA) miR-375 在调节饮食影响中的作用 miRNA 是对 EEC 生物学做出反应的短非编码 RNA 分子。 改变环境背景并在转录后水平调节基因表达。 miRNA 是多种生物途径的关键调节因子，包括肠上皮发育和 我们的实验室此前已证明 miR-375 在肠道干细胞 (ISC) 中高度富集。 沿着 EEC 谱系，其表达因长期高脂肪饮食而显着减少。 初步数据表明减肥手术后 ISC 中 miR-375 表达得到显着恢复， 与 EEC 丰度和循环肠道激素水平的增加相一致。 miR-375 在饮食引起的 EEC 的发病机制和改善过程中对 EEC 生物学产生特定的影响 拟议的研究将使用我们实验室的跨学科方法来检验这一假设。 建立了 miR-375 敲除 (375-KO) 小鼠群体以及尖端基因组和生物信息学 在目标 1 中，我将评估 miR-375 的丢失如何对饮食分布产生影响。 通过对小细胞进行高分辨率单细胞 RNA 测序 (scRNA-seq) 来分析不同的 EEC 亚型 来自长期喂食或高脂肪饮食的野生型 (WT) 和 375-KO 动物的肠隐窝和绒毛。 目标 2，我将确定 miR-375 如何促进手术诱导的 EEC 适应和代谢 第375章 借助这些单细胞数据集，我将通过生物信息学方法对减肥手术或对照手术后的小鼠进行 KO。 确定总体 EEC 丰度、亚型分布的上下文相关变化（与 循环肠道激素水平）和基因表达（包括候选 miR-375 靶点的鉴定）。 还将通过免疫组织化学测定和代谢在体内验证这些分子发现 总而言之，这些发现将进一步加深我们对体重和葡萄糖耐量等参数的理解。 EEC 调节的作用，并可能为肥胖及其合并症的治疗提供新的治疗靶点。

项目成果

The long-acting amylin/calcitonin receptor agonist ZP5461 suppresses food intake and body weight in male rats.

长效胰淀素/降钙素受体激动剂 ZP5461 可抑制雄性大鼠的食物摄入量和体重。

• DOI: 10.1152/ajpregu.00337.2020
• 发表时间: 2021
• 期刊: American journal of physiology. Regulatory, integrative and comparative physiology
• 作者: Stein,LaurenM;McGrath,LaurenE;Lhamo,Rinzin;Koch-Laskowski,Kieran;Fortin,SamanthaM;Skarbaliene,Jolanta;Baader-Pagler,Tamara;Just,Rasmus;Hayes,MatthewR;Mietlicki-Baase,ElizabethG
• 通讯作者: Mietlicki-Baase,ElizabethG