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.

项目摘要 /摘要 肠内分泌细胞（EEC）协调各种信号网络，以维持代谢稳态。 作为肠道上皮的罕见秘密细胞谱系，EEC感知并通过释放A响应腔刺激。 控制营养感的训练，食欲，血糖调节和能量平衡。饮食- 诱导的肥胖症和减肥手术与这些失调和恢复有关 荷尔蒙途径。此外，越来越多的药物策略已经出现 目标关键EEC信号传导途径治疗代谢疾病。但是，泛岩这些进步分子 进行EEC生物学的机制仍未完全定义。为了解决这个知识差距，该提议 旨在确定富含EEC的microRNA（miRNA），miR-375在控制饮食的作用中的作用 以及对EEC生物学的手术干预措施。 miRNA是响应的短而非编码的RNA分子 改变环境环境并在转录后调节基因表达。像这样， miRNA是无数生物途径的关键调节剂，包括肠上皮发育和 功能。我们的实验室以前已经证明，miR-375在肠道干细胞（ISC）中都高度富集 沿着EEC谱系，其表达通过慢性高脂饮食大大降低。另外，我们的 初步数据表明，在减肥手术后，ISC中miR-375的表达显着拯救， 与EEC抽象的增加和循环肠horse级水平相吻合。因此，我假设 miR-375在饮食引起的发病机理和改善期间对EEC生物学产生上下文特异性影响 肥胖。拟议的研究将使用我们的实验室的跨学科方法来检验这一假设 MiR-375敲除（375-KO）小鼠的固定菌落与尖端的基因组和生物学 技术。在AIM 1中，我将评估miR-375的损失如何对饮食特异性影响对分布 通过进行高分辨率的单细胞RNA测序（SCRNA-SEQ）的不同EEC亚型 来自Wildtype（WT）和375-KO动物的肠道加密和Villi喂了慢性食物或高脂饮食。在 AIM 2，我将确定miR-375如何贡献外科手术诱导的EEC适应和代谢 通过对饮食诱发的肥胖WT和375-的地穴和绒毛样品的SCRNA-seq分析进行改进 减肥手术或对照程序后的KO小鼠。使用这些单细胞数据集，我将生化 确定整体EEC抽象，亚型分布的依赖于上下文的变化（与 循环肠hor骨水平）和基因表达（包括鉴定候选miR-375靶标）。 还将通过免疫组织化学和代谢在体内验证这些分子发现 参数，例如体重和葡萄糖耐受性。总之，这些发现将进一步我们的理解 EEC调节的规定，可以为治疗肥胖症及其合并症提供新颖的治疗靶标。

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