Engineering Immuno-Glial-Neurovascular 3D-Brain-Chips with a Perfusable BBB for Accelerating Alzheimer’s Disease Drug Discovery and Translation

项目摘要。虽然阿尔茨海默病（AD）是第六大死亡原因，但仍然没有 市场上的药物治疗，减缓或停止AD中的神经元损伤。载脂蛋白-E4（APOE 4） 是散发性AD最强的遗传风险因素， 携带。重要的是，进一步阻碍有效治疗开发的是缺乏基于人类的模型， 可以概括AD病理学与所有的脑驻留细胞类型的免疫胶质细胞神经血管单位，帽， 关键细胞非自主效应，包括炎症和脂质失调，以及血脑 屏障（BBB）介导的营养物和治疗剂的转运。为了解决这些限制，在一个雄心勃勃的 在我博士后工作的第一阶段，我设计了一个新颖的大脑模拟矩阵 （NeuroMatrix），支持来自患者特异性诱导多能干细胞的所有7种脑细胞类型的共培养 具有成熟表型的细胞以形成多细胞整合脑模型（miBRAIN）。该模型概括了 APOE 4相关失调和AD病理学标志神经元过度兴奋、淀粉样蛋白过度兴奋、 此外，我已经开发了一种新的收缩最小化微流体， 在NeuroMatrix中自组装的神经血管单元内实现可灌注血管的器械。我提议 目的是：（目标1）利用miBRAIN平台构建多元化的“体外患者队列” APOE基因型与等基因系，以探测APOE 4特异性效应，并进行重要验证， 模型，因为我的目标是磨练这个平台技术的广泛实用性和（目标2）工程3D-miBRAIN- 通过将我的新型微流控装置与miBRAIN相结合， 在miBRAIN培养物中朝向完全可灌注的BBB，并利用它们来功能性地评估BBB渗透性。 在患者细胞系和APOE状态之间的能力和选择性。与项目官员协商后 考虑到我的长期目标，我决定进一步关注（目标3）研究最佳前进道路 部署这项技术以实现最大效用，包括制定商业计划和确定 潜在合作伙伴和重点应用程序。因此，这项工作可能会导致AD机制的重要见解 了解与APOE 4风险相关的其他神经退行性疾病的直接相关性， 开发一种经过充分验证的3D-miBRAIN-Chip平台技术，该技术是一种集成的免疫-神经胶质-神经血管- 具有患者特异性遗传学和仿生表型和功能的cular单元， CNS病理学的治疗进展。同时，它提供了重要的研究方法培训， ods（平台开发，生物信息学，脂质组学和炎症分析，专家合作指导， 导师和科学咨询委员会）和职业发展（沉浸式创业培训， 利基特定的课程，从专家共同导师和创业咨询委员会指导，追求 雄心勃勃的独立愿景），这将在启动我的独立职业生涯中产生显着的影响。

PROJECT SUMMARY. While Alzheimer’s Disease (AD) is the sixth leading cause of death, there is still no pharmacologic treatment on the market that slows or stops neuronal damage in AD. Apolipoprotein-E4 (APOE4) is the strongest genetic risk factor for sporadic AD and one that roughly a quarter of the general population carries. Importantly, further precluding effective therapeutic development is the lack of human-based models that can recapitulate AD pathology with all the brain-resident cell types in the immune-glial-neurovascular unit, cap- turing critical cell non-autonomous effects, including in inflammatory and lipid dysregulation, and blood brain barrier (BBB)-mediated transport of nutrients and therapeutics. To address these limitations, in an ambitious moonshot project in the first phase of my postdoctoral work, I have engineered a novel brain-mimetic matrix (NeuroMatrix) that supports the co-culture of all 7 brain cell types from patient-specific induced pluripotent stem cells with mature phenotypes to form a multi-cellular integrated brain model (miBRAIN). This model recapitulates APOE4-associated dysregulation and AD pathological hallmarks of neuronal hyperexcitability, amyloid accumu- lation, phosphorylated tau burden, etc. Further, I have developed a novel constriction-minimizing microfluidic device that enables perfusable vasculature within neurovascular units self-assembled in NeuroMatrix. I propose here to: (Aim 1) leverage the miBRAIN platform to construct a diverse “in vitro patient cohort” across APOE genotype with isogenic lines to probe APOE4-specific effects and conduct important validation of the model as I aim to hone this platform technology for broad utility and (Aim 2) engineer 3D-miBRAIN- Chips by combining my novel microfluidic devices with the miBRAIN and successively integrating each cell type towards a fully perfusable BBB within miBRAIN culture and harness them to functionally assess BBB perme- ability and selectivity across patient cell lines and APOE status. After consultation with the Program Officer and given my long-term goals, I have decided to further focus on (Aim 3) investigating the best path forward for deploying this technology for maximal utility, including development of a business plan and identification of potential partners and focus applications. This work thus could result in important insights for AD mechanistic understanding, of immediate relevance to other neurodegenerative diseases associated with APOE4 risk, while developing a well-validated 3D-miBRAIN-Chip platform technology that is an integrated immune-glial-neurovas- cular unit with patient-specific genetics and biomimetic phenotypes and functions that could be of great utility for therapeutic development across CNS pathologies. Simultaneously, it provides vital training in research meth- ods (platform development, bioinformatics, lipidomic and inflammation analyses, mentoring from expert Co- Mentors and Scientific Advisory Committee) and career development (immersive entrepreneurial training, niche-specific coursework, mentoring from expert Co-Mentors and Entrepreneurial Advisory Committee, pursuit of ambitious independent vision) that will make a marked difference in launching my independent career.