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

摘要 随着人口老龄化，对改进脑血管疾病研究方法的需求迅速增长 增加。与此同时，人们对再现血液生理特性的兴趣日益浓厚。 体外脑屏障（BBB），对基础和临床研究都有帮助。因此需要 标准化模型作为工具，帮助科学家更好地理解生理和病理机制 涉及脑血管疾病，包括阿尔茨海默病（AD）。拟议的研究旨在 开发一种多功能 BBB 模型，以三维方式概括 BBB 微血管网络 (μVN) (3D)微流控平台，并将该模型应用于AD发病机制研究。建立在我们广泛的一套 初步和相关数据，我们建议在细胞外基质（ECM）内重建 3D AD-BBB µVN 支架能够实现细胞间信号传导并以明确的方式暴露于生化梯度 微环境。 AD是一种进行性神经退行性疾病，其特征是认知功能退化和 β-淀粉样蛋白 (Aβ) 肽的沉积。我们之前报道过人类神经祖细胞过度表达 淀粉样β前体蛋白 (APP) 和早老素 1 (PSEN1) 基因中生长的家族性 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，该模型缺乏 BBB 成分对神经功能和 AD 发病机制至关重要。我们将开发3D- 在 BBB µVN 存在的情况下，在 3D 微流体平台中分化 AD 细胞（3D AD-BBB µVN 模型）。 我们将研究 AD 中 BBB 是如何被破坏的以及优化 BBB 是否可以改善 AD 进展。我们的 3D AD-BBB µVN 模型将适用于 BBB 相关疾病致病级联，例如 例如AD，以及在类似人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.