Pathophysiology of Diabetic Gastroparesis

糖尿病胃轻瘫的病理生理学

• 批准号: 10618295
• 负责人: GIANRICO FARRUGIA
• 金额: $ 55.41万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10618295
• 关键词: ATAC-seq; Address; Affect; Anti-Inflammatory Agents; Biological; Blood Cells; Cells; Complement; Complications of Diabetes Mellitus; Cytometry; Data; Development; Diabetes Mellitus; Diabetic mouse; Disease; Enhancers; Environment; Epigenetic Process; Equilibrium; Exposure to; Functional disorder; Gastric Emptying; Gastroparesis; Gene Expression; Genes; Genetic; Health; Immune; Infiltration; Inflammation Mediators; Inflammatory; Interstitial Cell of Cajal; Knowledge; Macrophage; Metabolism; Mononuclear; Mus; Obstruction; Oxidative Stress; Patients; Phagocytes; Phenotype; Play; Positioning Attribute; Prevention; Proteins; Role; Signal Pathway; Solid; Source; Stomach; System; Techniques; Technology; Testing; Tissues; Work; cell injury; cell type; cytokine; diabetic; diabetic gastroparesis; epigenomics; experimental study; gastrointestinal; improved; monocyte; paracrine; preservation; response; single-cell RNA sequencing; therapy development

PROJECT SUMMARY/ABSTRACT Diabetes mellitus is associated with multiple complications including diabetic gastroparesis. The systemic effects of diabetes modify all cells in the body but in common with other diabetic complications, gastroparesis does not develop in all patients. Delayed gastric emptying (GE) in the absence of obstruction defines gastroparesis and also occurs in some but not all diabetic mice. These observations suggest that together with genetics, convergence of systemic changes with local factors is necessary to develop delayed GE in diabetes. We previously have found that delayed GE that is associated with loss of interstitial cells of Cajal (ICC), which in turn is associated with fewer CD206+ muscularis propria macrophages (MPMs). MPMs play a central role in modifying the cellular content and biological environment of the GI muscularis propria in health and disease. The source of gastric MPMs and the mechanisms for activation of injurious molecules in MPMs by diabetes are not known. The overall hypothesis of this proposal is that the underlying central determinant of delayed gastric emptying in diabetes is the combination of a programmed activation response evoked in monocytes and the tissue environment of new monocyte-derived MPMs in the gastric muscularis propria. This will be tested in three specific aims (SA). SA1 will determine the effect of diabetes on circulating monocytes that primes them for infiltration and activation in gastric muscularis propria. SA2 will determine the origin of activated MPMs that populate the gastric muscularis propria following onset of diabetes and SA3 will determine how newly established and activated MPMs cause cellular injury in the diabetic stomach. The SAs are supported by extensive preliminary data. 1) We have found that systemic exposure to diabetes modifies the epigenomic landscape of circulating monocytes so that monocytes are poised to express a restricted complement of signaling pathways when they enter the tissue and become MPMs; 2) MPMs in the diabetic stomach are derived from infiltrating monocytes; 3) Diabetes results in the presence of new, distinct, discrete types of MPMs that are unique to the tissue; 4) Lineage-tagged, long-lived MPMs are not changed by diabetes; 5) Disruption of cellular metabolism in the gastric muscularis propria by diabetes activates monocyte-derived MPMs in close apposition to ICC to produce injurious cytokines, which cause damage to ICC and disrupted gastric function. To address the overall hypothesis we have developed techniques to isolate identified MPMs and monocytes, applied highly sophisticated technology for profiling genes by single cell RNASeq, ATAC-Seq, and proteins by mass cytometry (CyTOF), and determined changes in the epigenomic landscape in innate immune cells based on phenotypes identified in single cell studies. The preliminary data and the proposed experiments position us to identify the key steps that determine the responses to diabetes in innate immune cells that result in development of delayed GE. The new knowledge gained will be crucial to direct targeting of the mononuclear phagocyte system for prediction, prevention and treatment of diabetic gastroparesis.

项目摘要/摘要 糖尿病与包括糖尿病性胃轻瘫在内的多种并发症有关。系统性 糖尿病的作用改变了体内的所有细胞，但与其他糖尿病并发症，胃轻瘫相同 并非所有患者发展。在没有阻塞的情况下延迟胃排空（GE）定义 胃轻瘫，也发生在一些但不是所有糖尿病小鼠中。这些观察结果表明 遗传学，全身变化与局部因素的收敛是为了发展糖尿病中的延迟GE。 我们以前已经发现，延迟的GE与Cajal（ICC）的间隙细胞损失有关，该细胞损失 反过来又与较少的CD206+ Muscularis Proprias巨噬细胞（MPMS）有关。 MPM在 修改GI Muscularis Prestrias在健康和疾病中的细胞含量和生物学环境。 胃MPM的来源和糖尿病中MPM中有害分子激活的机制是 不知道。该提议的总体假设是延迟的基本中央决定因素 糖尿病中的胃排空是唤起的编程激活反应的组合 单核细胞和新单核细胞衍生的MPM的组织环境 普遍。这将在三个特定目标（SA）中进行测试。 SA1将确定糖尿病对循环的影响 单核细胞在胃muscularis proprias中浸润和激活。 SA2将确定 糖尿病和SA3发作后，激活的MPM填充了胃肌的过度 确定新建立和激活的MPM如何在糖尿病胃中导致细胞损伤。 SAS 由广泛的初步数据支持。 1）我们发现全身暴露于糖尿病 循环单核细胞的表观基因组景观，以表达有限的单核细胞 信号通路进入组织并成为MPM时的补充； 2）糖尿病患者中的MPM 胃是源自浸润的单核细胞。 3）糖尿病导致存在新的，独特的离散 组织独有的MPM类型； 4）糖尿病不会改变谱系标签的长期MPM； 5）糖尿病胃肌中的细胞代谢破坏会激活单核细胞来源 MPM密切与ICC息息下生产有害细胞因子，这会损害ICC并破坏 胃功能。为了解决总体假设，我们开发了隔离MPM的技术 和单核细胞，将高度复杂的技术应用于单细胞RNASEQ，ATAC-SEQ，ATAC-SEQ， 通过质量细胞仪（CytoF）和蛋白质的蛋白质，并确定了先天性表观基因组景观的变化 免疫细胞基于单细胞研究中鉴定的表型。初步数据和提议 实验将我们定位为确定确定先天免疫中对糖尿病的反应的关键步骤 导致GE延迟发展的细胞。获得的新知识对于直接定位至关重要 用于预测，预防和治疗糖尿病性胃轻瘫的单核吞噬细胞系统。