Human Microphysiology Systems Disease Model of Type 2 Diabetes Starting with Liver and pancreatic Islets

从肝、胰岛入手的2型糖尿病人体微生理系统疾病模型 超过3000万美国人患有糖尿病，约占成年人口的9.4%。额外的84 数以百万计的美国成年人患有糖尿病前期，这两种疾病每年造成的经济损失高达3220亿美元。这个 所有形式的糖尿病的根本原因是相对于代谢需求的胰岛素分泌不足。 虽然1型糖尿病(T1D)的β细胞由于自身免疫破坏而绝对丧失，但 2型糖尿病(T2D)的发病机制与既往的胰岛素抵抗密切相关 存在于许多组织，主要是肝脏、胰岛中的β细胞、白色脂肪组织和骨骼组织 肌肉。胰岛素抵抗和代谢后果因组织而异，更重要的是， 在人口上有很大的差异。此外，来自人体和模型生物体研究的证据 论证了器官串扰的重要性，包括肌肉因子、脂肪因子、肝因子和 炎性细胞的细胞因子以及miRNA的胞外转移在慢性粒细胞白血病的病理生理中的作用 糖尿病。人类和模式生物生理学的种间差异限制了 许多发现(例如来自转基因小鼠的研究)，例如来自β细胞的发现。所有这些都是成功的 有必要设计体外系统来研究允许器官串扰询问的人体生理学。 在人类微生理学系统(MPS)中理解T2D的病理生理学将有助于理解 疾病的进展，识别生物标志物，并制定治疗策略，以预防、减轻或 扭转与糖尿病相关的发病率，改善患者预后。我们的建议集中在以下两个方面 关键器官：肝脏和胰岛。我们将首先演示相关的生理学和 血管化肝腺泡细胞(VLAMPS)和血管胰岛MPS的病理生理 (VPANIS)使用原代人类细胞/组织(目标1)。MPS疾病模型的全部力量将利用患者- 器官中所有关键细胞的衍生成人IPSCs，包括KEY的实时荧光生物传感器 选定基因的生理参数和条件性击倒。我们的建议有一个战略计划，以 优化从UG3期的原代人类细胞到晚期IPSC来源的细胞的迁移 普遍定期审议阶段，包括合作整合综合方案领域的相关进展(目标2和4)。这个 人类原发的、基于细胞的MPS的初始使用将定义每个MPS的最佳正常和疾病指标 模型，以开始疾病的调查，并作为功能参考，以测试生理 IPSC衍生模型的相关性。我们将在功能上将vLAMP耦合到 VPANIS将测试来自胰岛素抵抗肝脏的因素可以加强β细胞功能障碍的假设 高血糖和高胰岛素血症的背景(目标3和4)。我们将使用我们的微生理学数据库 共享数据、协议、试剂、vLAMPS和vPANIS模型和结果的平台(目标5)。

Human Microphysiology Systems Disease Model of Type 2 Diabetes Starting with Liver and Pancreatic Islets Over 30 million Americans have diabetes, constituting about 9.4% of the adult population. An additional 84 million adult Americans have pre-diabetes, both amounting to an economic cost of $322 billion annually. The underlying cause of all forms of diabetes is an inadequate insulin secretion relative to the metabolic needs. While there is an absolute loss of beta cells in type 1 diabetes (T1D) due to an autoimmune destruction, the pathogenesis of type 2 diabetes (T2D) is much more heterogeneous with preceding insulin resistance being present in many tissues, principally the liver, β-cells in pancreatic islets, white adipose tissue and skeletal muscle. The insulin resistance and the metabolic consequences vary between tissues and more importantly, vary enormously in the population. Furthermore, evidence from human and model organism studies has demonstrated the importance of organ crosstalk including the role of myokines, adipokines, hepatokines and cytokines from inflammatory cells, as well as the exosomal transfer of miRNA in the pathophysiology of diabetes. Interspecies differences between human and model organism physiology limits the translatability of many findings (e.g. from transgenic mouse studies), such as those from beta cells. All of these make it necessary to devise in vitro systems to study human physiology that allow organ crosstalk interrogation. Understanding the pathophysiology of T2D in a human microphysiology system (MPS) will help understand the progression of the disease, identify biomarkers and develop therapeutic strategies that can prevent, mitigate or reverse the morbidity associated with diabetes and improve patient outcomes. Our proposal focuses on two of the critical organs: liver and pancreatic islets. We will first demonstrate the relevant physiology and pathophysiology in the vascularized liver acinus MPS (vLAMPS) and the vascularized pancreatic islets MPS (vPANIS) using primary human cells/tissue (Aim 1). The full power of MPS disease models will utilize patient- derived, adult iPSCs of all of the key cells in the organs and include real-time fluorescent biosensors of key physiological parameters and conditional knock-downs of selected genes. Our proposal has a strategic plan to optimize the migration from primary human cells in the UG3 phase to iPSC-derived cells in the later stages of the UH3 phase, including collaborative integration of relevant progress in the iPSC field (Aim 2 and 4). The initial use of human primary, cell-based MPS’s will define the optimal normal and disease metrics in each MPS model to begin the investigation of the disease and to serve as a functional reference to test the physiological relevance of the iPSC-derived models. We will functionally and then physically couple the vLAMPS to the vPANIS to test the hypothesis that factors from the insulin resistant liver can potentiate beta cell dysfunction in the context of hyperglycemia and hyperinsulemia (Aims 3 and 4). We will use our microphysiology database as a platform for sharing data, protocols, reagents, the vLAMPS and vPANIS models and results (Aim 5).