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

项目概要/摘要 肥胖相关疾病，特别是2型糖尿病（T2 DM），在美国持续增加， 据估计，全球有19亿超重成年人和超过6.5亿肥胖者。而 肥胖诱导的T2 DM的机制基础仍然是一个研究主题， 包括促炎环境和脂肪组织中脂解失调，导致水平升高 循环中游离脂肪酸与随后在多个组织中的脂质异位积累。的 营养过剩和促炎信号传导的组合反过来导致多个人的胰岛素抵抗。 组织损害肌肉和脂肪组织的葡萄糖摄取以及肝脏和胰岛细胞的葡萄糖释放 功能，最终导致明显的糖尿病。对这些关键组织之间复杂相互作用的研究 到目前为止，只能使用动物模型，这并不适合高通量 在关键代谢特征上接近并经常偏离人类，因此极大地阻碍了 发现胰岛素抵抗和T2 DM的治疗方法。在这里，我们建议制定一套基本的人类 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， 也就是说，肝脏（L）和脂肪（A）组织，以及胰岛素分泌，即，胰岛（I），与免疫 组件，即，巨噬细胞，使用互连微生理系统（MPS）。该LAI-MPS将 允许在正常组织的情况下对葡萄糖和胰岛素敏感性进行药理学询问 目前，该领域的研究主要集中在药物相互作用、脂质过载和慢性炎症方面，以解决以下主要的当前不足。在6 里程碑我们将从1）人类iPSC衍生物的产生和代谢表征 肝细胞、脂肪细胞、巨噬细胞-至2）开发优化的微流体装置， iPSC衍生的肝细胞、脂肪细胞和胰岛细胞-至3）建立芯片上胰岛素和葡萄糖敏感性 测定WAT和胰岛MPS。作为UH 3阶段的一部分，我们将开始MPS平台的集成， 4)肝脏和脂肪MPS与普通培养基的整合以及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的缩放和促炎性巨噬细胞的包含-最后6）整合胰岛，肝脏， WAT MPS并确定药理学和促炎调节对葡萄糖耐量的影响 和胰岛细胞功能。最终，这项颠覆性技术将使快速筛选药理学 和环境化合物对胰岛素敏感性的有益或有害作用，以及用于检测 药物遗传学相互作用

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.