Engineering a dynamic three-dimensional in vitro platform for the investigation of human Type 1 Diabetes immunopathogenesis

项目概要/摘要 1型糖尿病（T1 D）是一种自身免疫性疾病，由异常T细胞介导的靶向破坏 胰腺中产生胰岛素的β细胞，导致血糖调节丧失，长- 血管性和神经性共病的长期风险。尽管T1 D是研究最多的器官之一， 特定的自身免疫性疾病，旨在干预，预防或逆转这种疾病的各种策略， 疾病未能成功，由于不完全了解其确切的作用机制， 仅对系统性影响进行外围评估（例如，循环细胞因子变化，C肽水平） 可行缺乏对这些干预措施的机械理解，以及大量的时间和成本， 临床试验是改善治疗结果的一个重大障碍。为了解决这些重要的知识 空白，有大量的临床需要，以开发基于人类的离体系统，能够密切研究 胰岛与免疫细胞的相互作用以及环境因素对免疫细胞的影响 激活、归巢和细胞毒性。这一建议的主要假设是，一个小岛的发展- 免疫平台有可能通过研究激活途径，提供对T1 D的独特见解 和介入方法的筛选。因此，本提案的目标是设计、验证和 利用独特的体外3-D平台，通过汇聚 创新细胞与生物材料、原位成像和微生理系统（MPS）。目标1将寻求 建立并验证这个基于生物材料的三维共培养平台。为了验证系统，采用分层方法， 将采用从单抗原鼠模型细胞到人T1 D-抗原细胞的构建。一旦验证， 目标2将把这个平台转化为研究以人为中心的T1 D相关通路和干预措施。最后，Aim 3将寻求通过将3D材料转换为已建立的 微生理系统（MPS）平台，其将允许研究T细胞从流体微环境迁移。 微环境对β细胞的影响这一提议的结果应提供一个经过验证的有利工具 用于研究人类T1 D相关的病理生理学、干预和治疗。虽然拟议的外地 该平台的应用是T1 D，其他自身免疫性疾病可以从该工程台架中受益 platofrm，因为它们共享免疫细胞失调的同源标志。

PROJECT SUMMARY/ABSTRACT Type 1 Diabetes (T1D) is an autoimmune disease caused by aberrant T-cell mediated targeted destruction of insulin-producing beta cells in the pancreas, resulting in loss of blood glucose regulation, with increased long- term risks of vascular and neuropathic comorbidities. Despite the fact that T1D is one of the most studied organ- specific autoimmune diseases, the various strategies aimed at intervention, prevention, or reversal of this disease have failed to succeed due to incomplete knowledge about the precise mechanisms of their action, as only peripheral assessments of systemic impacts (e.g., circulating cytokine changes, C-peptide levels) are feasible. This lack of mechanistic understanding of these interventions, as well as substantial time and cost of clinical trials, is a profound obstacle in improving therapeutic outcomes. To address these significant knowledge gaps, there is a substantial clinical need to develop human-based ex vivo systems capable of intimately studying the interplay of islets and immune cells, as well as the contribution of environmental factors on immune cell activation, homing, and cytotoxicity. The primary hypothesis of this proposal is that the development of an islet- immune platform has the potential to provide unique insight into T1D, with investigation of activation pathways and screening of interventional approaches. Thus, the objective of this proposal is to engineer, validate, and utilize a unique in vitro 3-D platform for the interrogation of human T1D immunopathogenesis by converging innovative cells with biomaterials, in situ imaging, and microphysiological systems (MPS). Aim 1 will seek to establish and validate this 3D biomaterial-based co-culture platform. To validate the system, a tiered approach, building from single antigen murine model cells to human T1D-antigen cells, will be employed. Once validated, Aim 2 will translate this platform to study human-centric T1D-relevant pathways and interventions. Finally, Aim 3 will seek to integrate spatial and fluidic features by translating the 3D material to an established microphysiological system (MPS) platform, which will permit the study of T cell migration from a fluidic microenvironment to the beta cell niche. Results from this proposal should provide a validated and enabling tool for the study of human T1D-relevant pathophysiology, interventions, and therapeutics. While the proposed field of application for this platform is T1D, other autoimmune diseases can benefit from this engineered benchtop platofrm, as they share homologous hallmarks of immune cell dysregulation.