A 3D vascularized islet biomimetic to model type 1 diabetes

用于 1 型糖尿病模型的 3D 血管化胰岛仿生模型

• 批准号: 10449953
• 负责人: CHRISTOPHER C. W. HUGHES
• 金额: $ 100.96万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10449953
• 关键词: 3D; vascularized; islet; biomimetic; model

PROJECT SUMMARY/ABSTRACT Type 1 diabetes (T1D) is an autoimmune disease thought to be caused by immune-mediated destruction of the insulin-producing β-cells in the pancreatic islets. Studying the mechanisms that underlie β-cell destruction in humans with T1D has been challenging because most of the important immunological events occur before diagnosis. Furthermore, while rodent models have been informative in defining some aspects of T1D etiology, there are fundamental differences between the rodent and human pancreas with respect to islet architecture and vasculature, as well as between rodent and human immune systems. Additionally, important aspects of human T1D pathology are not replicated in the rodent models. Therefore, to fully understand human T1D pathophysiology, it is critical to develop a human model, where the interactions of all cells involved in the disease process (e.g. β-cells, endothelial cells (EC), innate and adaptive immune cells) can be studied in the context of normal islet architecture, including vasculature, stromal cells, and native islet matrix. Over the past three years through the NIH “Consortium on Human Islet Biomimetics”, our team (co-PI Sander: human induced pluripotent stem cells (hiPSC) and diabetes; co-PI Hughes: vascular biology and bioengineering; co-I Christman biomaterials and tissue engineering; co-I George microfluidics and transport) has developed a microfluidic-based platform in which primary human islets or hiPSC-derived islet-like clusters are supported by a network of perfused human microvessels. Our 3D vascularized islet micro-organ (VMO-I) platform allows for physiologic, microvessel-mediated delivery of nutrients, disease-relevant stimuli, or immune cells to the islets. We propose to leverage the unique features of our VMO-I platform to model the cell-cell interactions that occur in the islet niche during T1D pathogenesis, namely immune cell extravasation, tissue penetration, and migration as well as β-cell killing. For these studies co-I Teyton will provide expertise in T1D immunology. We propose to employ two distinct in vitro models: The first, developed in the UG3 phase, is non-autologous and comprised of primary human islets and vasculature from primary EC. Here, we will introduce either allogeneic lymphocytes (Aim G1) or islet donor-matched β-cell-reactive T cell clones (Aim G2) to establish parameters for modeling T cell extravasation and T cell-mediated β-cell killing. We will also work towards the goal of generating a VMO-I model entirely derived from hiPSC (Aim G3). The second model, developed in the UH3 phase, will be fully autologous, comprising β-cells, vasculature, and stromal cells derived from T1D patient hiPSC, which will be combined with autoreactive T cells isolated from blood of the same patient. By combining live-sensors and real-time imaging with molecular and biochemical assays, we will use these models to study how cells in the islet respond to T1D- relevant stressors, such as pro-inflammatory cytokines, hyperglycemia, and hypoxia, how immune cells and β- cells interact, and how β-cells are killed. Finally, we will demonstrate that the platform can be used to assess candidate therapies for efficacy with the long-term goal to utilize the platform to screen for new therapeutics.

项目摘要/摘要 1型糖尿病(T1D)是一种自身免疫性疾病，被认为是由免疫介导的 胰岛中产生胰岛素的β细胞。研究β细胞破坏的机制 患有T1D的人类一直具有挑战性，因为大多数重要的免疫学事件都发生在 诊断。此外，虽然啮齿动物模型在定义T1D病因学的某些方面提供了信息， 啮齿类动物和人类胰腺在胰岛结构和 血管系统，以及啮齿动物和人类免疫系统之间的关系。此外，人类的重要方面 在啮齿动物模型中，T1D病理不复制。因此，要充分理解人类的T1D 在病理生理学方面，开发一种人体模型是至关重要的，在这种模型中，参与疾病的所有细胞的相互作用 过程(例如β细胞、内皮细胞(EC)、先天免疫细胞和获得性免疫细胞)可在以下背景下研究 正常的胰岛结构，包括血管、基质细胞和天然胰岛基质。在过去三年中 通过美国国立卫生研究院“人类胰岛仿生联盟”，我们的团队(共同-Pi Sander：人类诱导多能性 干细胞(HiPSC)与糖尿病；合作者皮休斯：血管生物学和生物工程；合作者克里斯曼 生物材料和组织工程；co-i George微流体和运输)已经开发出一种基于微流体的 一种平台，其中主要的人类胰岛或由HiPSC衍生的类胰岛簇由灌流的网络支持 人体微血管。我们的3D血运胰岛微器官(VMO-I)平台允许生理学、 微血管介导的营养物质、疾病相关刺激或免疫细胞向胰岛的输送。我们建议 利用我们的VMO-I平台的独特功能来模拟发生在小岛上的细胞与细胞之间的相互作用 T1D发病过程中的生态位，即免疫细胞外渗、组织穿透、迁移和迁移 β--细胞杀戮。对于这些研究，co-i Teyton将提供T1D免疫学方面的专业知识。我们建议雇用两名 独特的体外模型：第一种模型是在UG3阶段发展起来的，是非自体的，由初级 原发EC的人胰岛和血管构筑。在这里，我们将介绍同种异体淋巴细胞(目标G1) 或胰岛供者匹配的β细胞反应性T细胞克隆(AIM G2)，以建立T细胞建模的参数 渗出和T细胞介导的β细胞杀伤。我们还将朝着生成VMO-I模型的目标努力 完全来源于HiPSC(目标G3)。第二个模型是在UH3阶段开发的，将完全自体， 包括来自T1D患者的β细胞、血管系统和基质细胞，这些细胞将与 从同一患者的血液中分离出自身反应性T细胞。通过将实时传感器和实时成像相结合 通过分子和生化分析，我们将使用这些模型来研究胰岛细胞对T1D- 相关的应激源，如促炎细胞因子、高血糖和低氧，免疫细胞和β- 细胞之间的相互作用，以及β细胞是如何被杀死的。最后，我们将演示该平台可用于评估 疗效的候选疗法，长期目标是利用该平台筛选新的疗法。