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.

项目摘要/摘要 肥胖相关疾病，尤其是2型糖尿病（T2DM），在美国不断增加 在全球范围内，全球估计有19亿超重成年人和超过6.5亿肥胖的人。尽管 肥胖引起的T2DM的机械基础仍然是一个调查的话题，即中心特征 包括促炎环境和脂肪组织中的脂解失调，导致水平升高 循环的游离脂肪酸，随后多次脂质的生态积累。这 养分过量和促炎信号的结合反过来导致多种胰岛素抵抗 肌肉和脂肪组织吸收葡萄糖的组织，肝脏以及ß细胞释放 功能，最终导致明显的糖尿病。询问这些关键组织之间的复杂相互作用 到目前为止，仅使用动物模型才有可能，这些模型不适合高通量 方法并经常偏离人类的主要代谢特征，从而极大地阻碍了努力 发现胰岛素抵抗和T2DM的治疗方法。在这里，我们建议开发一组人类 诱导多能干细胞（IPSC）衍生的葡萄糖和脂肪酸摄取/释放的钥匙代谢组织， 即肝（L）和脂肪（A）组织和胰岛素分泌，即胰岛（I），与免疫结合 组件，即巨噬细胞，使用互连的微生理系统（MPS）。这个LAI-MP会 在正常组织的背景下，允许对葡萄糖和胰岛素敏感性的药物询问 相互作用，脂质超负荷和慢性炎症，以解决以下主要当前缺口。在6中 里程碑我们将从1）人IPSC衍生的产生和代谢表征中进步 肝细胞，脂肪细胞，ß细胞和巨噬细胞 - 至2）开发优化的微流体设备 IPSC衍生的肝细胞，脂肪细胞和ß细胞 - 至3）建立片上胰岛素和葡萄糖敏感性 屁股，沃特和小岛议员。作为UH3阶段的一部分，我们将通过 4）将肝脏和脂肪MP与常见培养基的整合，并使用在线测定胰岛素灵敏度 传感器 - 和5）使用肝脏和WAT MPS来生成和定量胰岛素抵抗性 WAT MPS的缩放和促炎性巨噬细胞的包含，最后6）综合胰岛，肝脏和 WAT MPS并确定药物和促炎调节对葡萄糖耐量的影响 和ß细胞功能。最终，这种破坏性技术将使药物快速筛查 和环境化合物，可对胰岛素敏感性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.