Alzheimer's disease-replicated brain microphysiological system to model AD physiopathology and its influenceon gliovasculature and immune system

摘要 阿尔茨海默病的两个特征是细胞外淀粉样斑块和细胞内淀粉样斑块堆积 神经原纤维缠绕，扰乱大脑功能。最近的证据表明，脑功能障碍 血管系统和免疫细胞过度活跃也在AD的进展中起着关键作用。在体外没有合适的 可靠地复制神经-神经胶质血管-免疫系统的生理病理或相互作用的模型 人类。此外，无论是基于大脑切片的分析，还是动物模型，都不能复制人类的全部光谱 阿尔茨海默病的神经病理和相关的胶质血管偶联炎症特征。早发性家族性 AD(FAD)是研究AD各个方面的有用和有代表性的模型，因为它是由一种 至少三个已知基因中的一个发生突变：早老素1、早老素2和淀粉样前体蛋白(APP)。 我们已经开发了一个FAD-大脑微生理系统(FAD-BMPS)来模拟健康和FAD-BMPS。 与脑源性细胞外基质(ECM)相关的神经元组织，并检测其生成 阿尔茨海默病的基本病理特征和特点。在一项初步研究中，我们建立了神经胶质血管- 整合神经元、星形胶质细胞、小胶质细胞和内皮细胞构建健康BMPS的免疫单位 分离的组织：脑组织和无膜血管。我们的概念验证研究也 论证了AD标志物产生的可行性及其对血管和炎症反应的影响。 我们建议开发一种3D无膜微流控FAD-BMPS，并通过以下方式验证AD的生理病理学 1)患者IPSC来源的FAD神经元组织与患者来源的ECM的集成，2)基于微流控的 无膜神经胶质血管系统；3)驻留和循环免疫细胞。我们将使用FAD神经元 并研究AD标志的产生，包括神经细胞中磷酸化tau沉积和淀粉样β蛋白 在患者来源的细胞外基质(ECM)中的积聚，以及淀粉样β蛋白的转运，吸收， 以及它的中介毒性。我们将比较胶质血管功能障碍和过度活跃的炎症反应 FAD-BMPS与来自健康BMPS的BMPS进行比较，评估AD病理恶化，并测试 改善FAD的现有药物和研究药物。这个创新的项目将结合优雅的 基于微流控技术的高通量、高含量脑内复杂相互作用成像能力 AD中的组织。这项研究最关键的方面是使用无膜神经胶质血管免疫。 系统以可重现的方式通常可应用于AD，提供现实的和临床相关的 数据，为药物筛选和个性化用药提供平台。皮耶洪云，一位 北卡罗来纳农工州立大学生物工程师，在脑微生理系统方面有丰富的经验 开发，由临床医生和干细胞科学家组成的团队提供支持。如果成功，这项BMPS将为 作为模拟AD的平台，减少动物使用，填补体外和体内之间的科学空白 模型，以及加速药物筛选和发现。

Abstract Two hallmarks of Alzheimers Disease are accumulation of extracellular amyloid plaques and intracellular neurofibrillary tangles, which disrupt brain function. Recent evidence indicates that dysfunction of cerebral vasculature and immune cell hyperactivity also play key roles in AD progression. There is no suitable in vitro model that reliably replicates the physiopathology or interaction of the neuro-gliovascular-immune system in humans. In addition, neither brain slice-based assays nor animal models replicate the full spectrum of human neuropathology and associated gliovascular-coupled inflammatory characteristics of AD. Early-onset familial AD (FAD) is a useful and representative model for studying various aspects of AD, since it is caused by a mutation in one of at least three knowns genes, presenilin 1, presenilin 2, and amyloid precursor protein (APP). We have developed a FAD-Brain MicroPhysiological System (FAD-BMPS) to model both healthy and FAD- relevant neuronal tissue with brain-derived extracellular matrix (ECM), and have tested the generation of essential pathological features and hallmarks of AD. In a pilot study, we established a neuro-gliovascular- immune unit of healthy BMPS by integrating neurons, astrocytes, microglia, and endothelial cells into two separate tissues: brain tissue and a membrane-free blood vessel. Our proof-of concept study also demonstrated the feasibility of AD hallmark generation and its impact on vascular and inflammatory responses. We propose to develop a 3D membrane-free microfluidic FAD-BMPS and to validate AD physiopathology by integrating 1) patient iPSC-derived FAD neuronal tissue with patient-derived ECM, 2) a microfluidic-based membrane-free gliovascular system, and 3) resident and circulating immune cells. We will use FAD-neurons and study AD hallmark generation, including phosphorylated tau deposit in neural cell and amyloid beta accumulation in the patient-derived extracellular matrix (ECM), as well as amyloid beta transport, absorption, and its mediated toxicity. We will compare gliovascular dysfunction and overactive inflammatory response in a FAD-BMPS with those from a healthy BMPS, assess AD pathology exacerbation, and test the efficacy of existing and investigational drugs in ameliorating FAD. This innovative project will combine the elegance of microfluidics-based high-throughput and high-content imaging capability with the complex interactions of brain tissue in AD. The most critical aspect of this study is employing a membrane-free neuro-gliovascular-immune system in a reproducible manner that can be generally applied to AD, providing realistic and clinically relevant data, and offering a platform for drug screening and personalized medicine. The PI Yeohung Yun, a bioengineer at NC A&T State University with considerable experience with brain microphysiological system development, is supported by a team of clinicians and stem cell scientists. If successful, this BMPS will serve as a platform for modeling AD, reducing animal use, filling the scientific gap between in vitro and in vivo models, and accelerating drug screening and discovery.