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

项目摘要/摘要 肥胖相关疾病，特别是2型糖尿病(T2 DM)在美国和 据估计，全球有19亿成年人超重，超过6.5亿人肥胖。而当 肥胖导致的2型糖尿病的机制基础仍然是一个研究的主题，主要特征 包括促炎环境和脂肪组织中失调的脂解导致水平升高 循环中的游离脂肪酸，随后脂质在多个组织中异位堆积。这个 营养过剩和促炎信号的结合反过来导致多发性硬化患者的胰岛素抵抗 影响肌肉和脂肪组织摄取葡萄糖并影响肝脏和肝细胞释放葡萄糖的组织 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询问 到目前为止，只有使用动物模型才有可能，动物模型不适合高吞吐量 方法，并经常偏离人类的关键代谢特征，从而极大地阻碍了努力 寻找治疗胰岛素抵抗和2型糖尿病的方法。在这里，我们建议开发一套基本的人类 诱导多能干细胞(IPSC)衍生的关键代谢组织，用于葡萄糖和脂肪酸的吸收/释放； 即肝脏(L)和脂肪(A)组织，以及胰岛素分泌，即胰岛(I)，与免疫相结合 使用互连的微生理系统(MPS)，即巨噬细胞。这位立马议员会 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 相互作用、脂质过载和慢性炎症，以解决当前的以下主要不足。在6年内 里程碑我们将从1)人IPSC的产生和代谢特性中取得进展 肝细胞、脂肪细胞、细胞和巨噬细胞-2)优化的微流控装置的开发 IPSC来源的肝细胞、脂肪细胞和？细胞3)建立芯片上的胰岛素和葡萄糖敏感性 检测MPS、WAT和胰岛MPS。作为UH3阶段的一部分，我们将通过以下方式开始整合MPS平台 4)肝脏和脂肪MPS与普通介质的集成和使用在线测定胰岛素敏感性 传感器-以及5)使用肝脏和Wat MPS产生和定量下列胰岛素抵抗 Wat MPS的鳞片和促炎巨噬细胞的包含-最终将胰岛、肝脏和 WAT MPS和测定药理和促炎调节对糖耐量的影响 和？细胞函数。最终，这种颠覆性的技术将使快速筛选药理作用成为可能。 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.