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 糖尿病与多种并发症有关，包括糖尿病胃轻瘫。系统性的 糖尿病的影响改变了体内的所有细胞，但与其他糖尿病并发症胃轻瘫有共同之处 并不是在所有患者中都会发生。无梗阻的胃排空延迟(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，与 遗传学上，全身改变与局部因素的趋同是糖尿病发生延迟性GE所必需的。 我们先前已经发现与Cajal间质细胞(ICC)丢失相关的延迟性GE，它 反过来，CD206+固有肌巨噬细胞(MPM)也较少。MPM在以下方面发挥了核心作用 在健康和疾病中改变胃肠道固有肌的细胞内容和生物环境。 胃MPM的来源和糖尿病激活MPM中损伤分子的机制如下 不知道。这一提议的总体假设是，延迟的潜在中央决定因素 糖尿病患者的胃排空是一种程序性激活反应的组合 单核细胞与胃肌单核细胞来源的新型MPM的组织环境 专有的。这将在三个具体目标(SA)中进行测试。SA1将决定糖尿病对循环的影响 单核细胞为胃固有肌的渗透和激活做好准备。SA2将决定 糖尿病发作后胃固有肌中激活的MPM的来源和SA3将 确定新建立和激活的MPM如何导致糖尿病胃中的细胞损伤。美国航空公司 都得到了大量初步数据的支持。1)我们发现，全身暴露在糖尿病环境中会改变 循环单核细胞的表观基因组图谱，使单核细胞能够表达限制性的 当它们进入组织并成为MPM时，信号通路的补充；2)糖尿病患者的MPM 胃来源于浸润性单核细胞；3)糖尿病导致新的、明显的、离散的 组织特有的MPM类型；4)有谱系标记的、寿命长的MPM不会因糖尿病而改变； 5)糖尿病破坏胃固有肌细胞代谢，激活单核细胞来源 与ICC紧密相对的MPM产生有害的细胞因子，从而对ICC造成损害并扰乱 胃功能。为了解决这一总体假设，我们开发了分离鉴定的MPM的技术 和单核细胞，应用高度复杂的技术通过单细胞RNAseq，ATAC-Seq， 和蛋白质，并确定了先天的表观基因组图谱的变化 基于单细胞研究中确定的表型的免疫细胞。初步数据和建议的 实验使我们能够确定在先天免疫中决定糖尿病反应的关键步骤 导致发育迟缓的GE的细胞。所获得的新知识将对直接瞄准 单核巨噬细胞系统用于预测、预防和治疗糖尿病胃轻瘫。