Microphysiological systems to interrogate the Islet-Liver-Adipose Axis in normal physiology and Type-2 Diabetes Mellitus

用于询问正常生理和 2 型糖尿病中的胰岛-肝脏-脂肪轴的微生理系统

• 批准号: 10462610
• 负责人: KEVIN Edward HEALY
• 金额: $ 228.78万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-20 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10462610
• 关键词: Address; Adipocytes; Adipose tissue; Animal Model; Beta Cell; Biological Assay; Cell Line; Cell physiology; Cells; Characteristics; Chronic; Coculture Techniques; Complex; Detection; Development; Devices; Diabetes Mellitus; Disease; Fatty Acids; Fatty Liver; Fatty acid glycerol esters; Functional disorder; Generations; Glucose; Glucose tolerance test; Hepatocyte; Hormonal; Human; Immune; Impairment; Inflammation; Inflammatory; Insulin; Insulin Resistance; Investigation; Lipids; Lipolysis; Liver; Metabolic; Metformin; Microfluidic Microchips; Modeling; Muscle; Non-Insulin-Dependent Diabetes Mellitus; Nonesterified Fatty Acids; Normal tissue morphology; Nutrient; Obesity; Pharmacogenetics; Pharmacology; Phase; Physiology; Protocols documentation; Rapid screening; Signal Transduction; Steatohepatitis; System; Technology; Testing; Tissues; chromatin immunoprecipitation; cytokine; design; glucose sensor; glucose tolerance; glucose uptake; human stem cells; induced pluripotent stem cell; inflammatory milieu; inflammatory modulation; insulin secretion; insulin sensitivity; insulin sensitizing drugs; insulin tolerance; islet; macrophage; microphysiology system; nonalcoholic steatohepatitis; novel; obese person; overweight adults; rapid detection; response; sensor; stem cell derived tissues; uptake

PROJECT SUMMARY/ABSTRACT Obesity-related disorders, particularly type-2 diabetes mellitus (T2DM), continuously increase in the US and worldwide with an estimated 1.9 billion overweight adults and over 650 million obese individuals globally. While the mechanistic underpinnings of obesity-induced T2DM remain a topic of investigation, central features include a pro-inflammatory environment and dysregulated lipolysis in adipose tissue leading to elevated levels of circulating free fatty acids with subsequent ectopic accumulation of lipids in multiple tissues. The combination of nutrient excess and pro-inflammatory signaling in turn results in insulin resistance in multiple tissues impairing glucose uptake by muscle and adipose tissue and release by the liver as well as ß-cell function, ultimately resulting in overt diabetes. Interrogation of the complex interplay between these key tissues has, thus far, only been possible using animal models, which do not lend themselves to high-throughput approaches and frequently deviate from humans in key metabolic features, thus greatly impeding efforts to discover treatments for insulin resistance and T2DM. Here we propose to develop an essential set of human induced pluripotent stem cell (iPSC)-derived key metabolic tissues for glucose and fatty acid uptake/release, i.e., liver (L) and adipose (A) tissue, and insulin secretion, i.e., islets (I), in conjunction with an immune component, i.e., macrophages, using interconnected microphysiological systems (MPS). This LAI-MPS will allow for the pharmacological interrogation of glucose and insulin sensitivity in the context of normal tissue interactions, lipid overload and chronic inflammation to address the following major current shortfalls. In 6 milestones we will progress from the 1) generation and metabolic characterization of human iPSC-derived hepatocytes, adipocytes, ß-cells and macrophages – to 2) Development of optimized microfluidic devices for iPSC-derived hepatocytes, adipocytes and ß-cells – to 3) Establish on-chip insulin and glucose sensitivity assays for, WAT and islet MPS. As part of the UH3 phase we will then begin integration of MPS platforms by 4) integration of liver and fat MPS with common medium and determination of insulin sensitivity using in-line sensors – and 5) Use liver and WAT MPS for the generation and quantitation of insulin resistance following scaling of WAT MPS and inclusion of pro-inflammatory macrophages – and finally 6) integrate islet, liver, and WAT MPS and determine impact of pharmacological and pro-inflammatory modulation on glucose tolerance and ß-cell function. Ultimately, this disruptive technology will enable the rapid screening of pharmacological and environmental compounds for beneficial or detrimental effects on insulin sensitivity and for the detection of pharmacogenetic interactions.

项目概要/摘要 肥胖相关疾病，特别是 2 型糖尿病 (T2DM) 在美国持续增加 全球估计有 19 亿成年人超重，超过 6.5 亿肥胖者。尽管 肥胖引起的 T2DM 的机制基础仍然是研究的主题，核心特征 包括促炎环境和脂肪组织中的脂解失调，导致水平升高 循环游离脂肪酸的减少，随后脂质在多个组织中异位积累。这 营养过剩和促炎信号传导的结合反过来会导致多种胰岛素抵抗 组织损害肌肉和脂肪组织对葡萄糖的摄取以及肝脏和β细胞的释放 功能，最终导致明显的糖尿病。探究这些关键组织之间复杂的相互作用 迄今为止，只能使用动物模型，而动物模型不适合高通量 方法，并且在关键代谢特征上经常偏离人类，从而极大地阻碍了 发现胰岛素抵抗和 T2DM 的治疗方法。在这里，我们建议开发一套基本的人类 诱导多能干细胞（iPSC）衍生的葡萄糖和脂肪酸摄取/释放的关键代谢组织， 即肝脏 (L) 和脂肪 (A) 组织，以及胰岛素分泌，即胰岛 (I)，与免疫结合 使用互连的微生理系统（MPS）的组件，即巨噬细胞。该 LAI-MPS 将 允许在正常组织的背景下对葡萄糖和胰岛素敏感性进行药理学询问 相互作用、脂质超载和慢性炎症，以解决当前的以下主要不足。 6年 我们将从 1) 人类 iPSC 衍生的生成和代谢表征开始取得进展 肝细胞、脂肪细胞、β 细胞和巨噬细胞 – 2) 开发优化的微流体装置 iPSC 衍生的肝细胞、脂肪细胞和 ß 细胞 – 3) 建立片上胰岛素和葡萄糖敏感性 WAT 和胰岛 MPS 的测定。作为 UH3 阶段的一部分，我们将开始整合 MPS 平台 4) 将肝脏和脂肪 MPS 与普通培养基整合并使用在线测定胰岛素敏感性 传感器 - 和 5) 使用肝脏和 WAT MPS 来生成和定量胰岛素抵抗 WAT MPS 的缩放和促炎巨噬细胞的包含 - 最后 6) 整合胰岛、肝脏和 WAT MPS 并确定药理和促炎调节对葡萄糖耐量的影响 和β细胞功能。最终，这种颠覆性技术将实现药理学的快速筛选 和环境化合物对胰岛素敏感性的有益或有害影响以及检测 药物遗传学相互作用。

项目成果

Multiplexed microfluidic platform for stem-cell derived pancreatic islet β cells.

• DOI: 10.1039/d2lc00468b
• 发表时间: 2022-11-08
• 期刊: LAB ON A CHIP
• 影响因子: 6.1
• 作者: Goswami, Ishan;de Klerk, Eleonora;Carnese, Phichitpol;Hebrok, Matthias;Healy, Kevin E.
• 通讯作者: Healy, Kevin E.