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破坏引起的 胰岛中产生胰岛素的β细胞。研究基于β细胞破坏的机制 患有T1D的人受到挑战，因为大多数重要的免疫事件发生在之前 诊断。此外，虽然啮齿动物模型在定义T1D病因的某些方面方面具有丰富的信息，但 啮齿动物和人类胰腺之间在胰岛结构和人类胰腺之间存在根本差异 脉管系统以及啮齿动物和人类免疫系统之间。另外，人类的重要方面 在啮齿动物模型中未复制T1D病理学。因此，要完全了解人类T1D 病理生理学，开发人类模型至关重要，其中所有参与该疾病的细胞的相互作用 过程（例如β细胞，内皮细胞（EC），先天性和适应性免疫小球）可以在 正常的胰岛结构，包括脉管系统，基质细胞和天然胰岛基质。在过去的三年中 通过NIH的“人类胰岛仿生体联盟”，我们的团队（Co-Pi Sander：人类诱发的多能 干细胞（HIPSC）和糖尿病； Co-Pi Hughes：血管生物学和生物工程； Co-i Christman 生物材料和组织工程； Co-I George Microfluidics and Transport）已经开发了基于微流体的 主要的人类胰岛或HIPSC衍生的胰岛样集群的平台由灌注网络支持 人类微血管。我们的3D血管化胰岛微孔器（VMO-I）平台允许生理， 微血管介导的养分，疾病相关的刺激或免疫细胞的递送到胰岛。我们建议 为了利用我们的VMO-I平台的独特功能来建模胰岛中发生的细胞电池相互作用 在T1D发病机理中的生态位，即免疫外渗出，组织穿透和迁移以及 β细胞杀戮。对于这些研究，Co-I Teyton将提供T1D免疫学专业知识。我们建议雇用两个 独特的体外模型：在UG3阶段开发的第一个是非自动的，并且是主要的 主要EC的人类胰岛和脉管系统。在这里，我们将引入同种异体淋巴细胞（AIM G1） 或胰岛供体匹配的β-cell反应性T细胞克隆（AIM G2）以建立建模T细胞的参数 渗出和T细胞介导的β细胞杀伤。我们还将致力于生成VMO-I模型的目标 完全来自HIPSC（AIM G3）。在UH3阶段开发的第二个模型将完全自体， 来自T1D患者HIPSC的β细胞，脉管系统和基质细胞将与该细胞合并 从同一患者的血液中分离出的自动反应性T细胞。通过结合实时传感器和实时成像 通过分子和生化测定，我们将使用这些模型来研究胰岛中的细胞对T1D-的反应 相关的压力源，例如促炎性细胞因子，高血糖和缺氧，免疫细胞和β- 细胞相互作用，以及如何杀死β细胞。最后，我们将证明该平台可用于评估 候选疗法提高效率的长期目标，以利用平台筛选新的治疗疗法。