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

项目摘要/摘要 1型糖尿病(T1D)是一种自身免疫性疾病，由异常的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，长时间血糖增加。 血管和神经性合并症的长期风险。尽管T1D是被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 疾病由于对其作用的确切机制的不完全了解而失败，如 只有对全身影响的外围评估(例如，循环细胞因子变化、C肽水平)才是 可行。缺乏对这些干预措施的机械性理解，以及大量的时间和成本 临床试验是改善治疗结果的一个严重障碍。要解决这些重要的知识 有很大的临床需求，需要开发能够深入研究的人体体外系统 胰岛与免疫细胞的相互作用及环境因素对免疫细胞的贡献 激活、归巢和细胞毒性。这项提议的主要假设是，一个小岛的发展- 免疫平台有可能通过对激活途径的研究来提供对T1D的独特见解 以及对介入方法的筛选。因此，本提案的目标是设计、验证和 利用独特的体外三维平台研究人类T1D免疫致病机制 具有生物材料、原位成像和微生理系统(MPS)的创新细胞。目标1将寻求 建立并验证了这个基于3D生物材料的共培养平台。为了验证该系统，采用分层方法， 将采用从单一抗原小鼠模型细胞到人T1D抗原细胞的构建。一旦得到验证， Aim 2将翻译这一平台来研究以人类为中心的T1D相关途径和干预。最后，瞄准 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，将允许研究T细胞从流体中的迁移 微环境对β细胞的影响。此建议书的结果应提供经过验证的支持工具 用于人类T1D相关病理生理学、干预和治疗的研究。虽然拟议的领域 这个平台的应用是T1D，其他自身免疫性疾病也可以从这个工程化的平台中受益 Platofrm，因为它们具有免疫细胞失调的同源特征。

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.