A Neurovascular Microphysiological System

项目摘要/摘要 最近，儿童神经发育障碍(NDDS)的发病率急剧增加。 例如，自闭症谱系障碍从20世纪80年代的每2000人中有1人增加到最近报告的 根据疾控中心的报告，每68人中就有1人。此外，因接触寨卡病毒而导致的原发性小头畸形症病例有 引起了全球的关注，沙利度胺和产前酒精暴露造成的出生缺陷突出了 开发合适的模型系统以了解、检测和治疗这些疾病的原因的重要性。 血管组织发育的重要性及其对疾病病理学的贡献往往被忽视， 因为重点放在神经和神经胶质细胞类型的作用上。然而，一些神经发育和 神经退行性疾病与血管畸形直接相关，包括小头畸形， 脑部海绵状畸形、自闭症、阿尔茨海默病、亚历山大病和雷特综合征。这个 大脑发育在神经发育的早期阶段是最脆弱的，也是 脆弱性是发育中的大脑从相关的神经周围血管丛形成的初始血管化 (PNVP)，血脑屏障尚未完全形成。到目前为止，还没有人的模型可以模仿 在这一关键阶段的脑血管系统。我们建议生成一种神经血管微生理系统(NV- MPS)，模拟发育中大脑的初始血管形成，并改进模型以支持长期 一种经过改造的可复制脑器官的长期维护。拟议的研究将实现三个具体目标 目标。具体目标1将使用无管状管创建一个正在开发的大脑微生理系统(MPS) 微流控设备、合成水凝胶和人类PSC来源的细胞类型。《特定目标2》将生成模型 并测量已确定的分子、细胞和组织水平的输出 是神经周围血管丛形成、神经血管整合和随后的皮质的指示物 成熟。特指目标3将测量和刺激NV-MPSS的神经元电生理功能 使用集成的蜂窝报告器和光学透明的嵌入电极。完成这项研究 将提供一种简单、健壮和可靠的NV-MPS，它将为研究初始血管形成提供手段 神经血管对神经系统疾病的贡献以及测试其效果的方法 在神经发育的早期阶段暴露于异种生物，最终将创造出可灌流的血管系统 来喂养和维持工程神经有机体。

PROJECT SUMMARY/ABSTRACT There has recently been a dramatic increase in incidences of neurodevelopment disorders (NDDs) in children. For instance, autism spectrum disorders have increased from 1 in 2000 people in 1980s to the recently reported 1 in 68 as reported by the CDC. In addition, instances of primary microcephaly due to Zika virus exposure have gained global attention, and birth defects caused by thalidomide and prenatal alcohol exposures highlight the importance of developing suitable model systems to understand, detect, and treat causes of these disorders. The importance of vascular tissue development and its contributions to disease pathologies are often overlooked, as the focus is instead on the role of neural and glial cell types. However, a number of neurodevelopmental and neurodegenerative disorders are directly associated with vascular malformations, including microcephaly, cerebral cavernous malformations, autism, Alzheimer’s disease, Alexander’s disease, and Rett syndrome. The developing brain is at its most vulnerable during the early stages of neurodevelopment, and one key point of vulnerability is the initial vascularization of the developing brain from the associated perineural vascular plexus (PNVP), where the blood brain barrier has yet to fully form. There is to date no human model that mimics the brain vasculature at this critical stage. We propose to generate a neurovascular microphysiological system (NV- MPS) that mimics the initial vascularization of the developing brain and to progress the model to support the long term maintenance of an engineered reproducible brain organoid. The proposed studies will achieve three specific aims. Specific Aim 1 will create a developing brain microphysiological system (MPS) using a tubeless microfluidics device, synthetic hydrogels and human PSC-derived cell types. Specific Aim 2 will generate models of a diseased developing brain and measure molecular, cellular and tissue level outputs that have been identified to be indicators of perineural vascular plexus formation, neurovascular integration and subsequent cortical maturation. Specific Aim 3 will measure and stimulate neuronal electrophysiological function in the NV-MPSs using integrated cellular reporters and optically transparent embedded electrodes. Completion of this research will provide a simple, robust and reliable NV-MPS that will provide the means to study the initial vascularization of the developing brain, the neurovascular contributions to neurological disorders and a means to test the effect of xenobiotic exposures at the early stages of neural development and finally will create a perfusable vasculature that feeds and maintains engineered neural organoids.