Development of 3D vascularized model of Blood Brain Barrier and its application to Alzheimer disease research

摘要 随着人口的老龄化，对研究脑血管疾病的改进方法的需求迅速增加 增加了。与此同时，人们对复制血液的生理特性的兴趣也在迅速增长- 体外脑屏障(BBB)，这对基础和临床研究都有帮助。因此，存在这样的需求 标准化模型作为帮助科学家更好地了解生理和病理机制的工具 涉及脑血管疾病，包括阿尔茨海默病(AD)。拟议的研究旨在 开发一种通用的血脑屏障模型，在三维模型中概括血脑屏障微血管网络(µVNS) (3D)微流控平台，并将该模型应用于AD发病机制的研究。建立在我们广泛的 初步和相关数据，我们建议在细胞外基质(ECM)内重建3D AD-BBB？VNS 支架，使细胞间信号和暴露在明确定义的生化梯度 微环境。 AD是一种进行性神经退行性疾病，其特征是认知功能恶化和 β-淀粉样蛋白(A-β)多肽沉积。我们之前曾报道过人类神经前体细胞过度表达 人淀粉样蛋白前体蛋白(APP)和早老素1(PSEN1)基因的家族性AD(FAD)突变 3D培养系统成功地概括了AD病理(3D AD培养模型)。然而，这种模式缺乏 血脑屏障成分，这是神经功能和AD发病机制的关键。我们将开发3D- 在BBB？VNS(3D AD-BBB？VN模型)存在的情况下，3D微流体平台中的差异化AD细胞。 我们将调查血脑屏障在AD中是如何被破坏的，以及优化血脑屏障是否可以改善AD 进步。我们的3D AD-BBB？VN模型将适用于两种与BBB相关的疾病致病级联，例如 作为AD，以及在类似人类大脑的环境中发现药物。

SUMMARY As the population grows older, the demand for improved methods to study cerebrovascular diseases has rapidly increased. At the same time, there has been burgeoning interest in reproducing physiological properties of blood- brain-barrier (BBB) in vitro that can be helpful in both basic and clinical studies. A need therefore exists for standardized models as tools to help scientists better understand the physiological and pathological mechanisms involved in cerebrovascular diseases, including Alzheimer’s disease (AD). The proposed studies are aimed at developing a versatile BBB model that recapitulates BBB microvascular networks (µVNs) in a three dimensional (3D) microfluidic platform, and apply this model to AD pathogenesis research. Building on our extensive set of preliminary and related data, we propose to recreate 3D AD-BBB µVNs within extracellular matrix (ECM) scaffolds that enable intercellular signaling and exposure to biochemical gradients in a well-defined microenvironment. AD is a progressive neurodegenerative disease which is characterized by deterioration of cognitive function and deposition of β-amyloid (Aβ) peptides. We previously reported that human neural progenitor cells overexpressing Familial AD (FAD) mutations in the amyloid-β precursor protein (APP) and presenilin 1 (PSEN1) genes grown in a 3D culture system successfully recapitulate AD pathologies (3D AD culture model). However, this model lacks BBB components, which are critical to neurological function and AD pathogenesis. We will develop 3D- differentiated AD cells in a 3D microfluidic platform in the presence of the BBB µVNs (3D AD-BBB µVN model). We will investigate how the BBB is disrupted in AD and whether optimizing the BBB can ameliorate AD progression. Our 3D AD-BBB µVN model will be useful for both BBB-related disease pathogenic cascades, such as AD, and drug discovery in a human brain-like environment.