Miniaturized AD/ADRD Microphysiological Systems Platform for High-throughput Screening

摘要 阿尔茨海默病（AD）的特征在于b-淀粉样蛋白（Ab）积累，神经元缠结（NFT）， 神经炎症和广泛的神经元和突触损失。到目前为止，还没有可用的治疗方法， 尽管免疫疗法即，勒坎莫布信守承诺。物种差异是 将动物模型中发现的治疗方法转化为人类脑特异性疾病，例如，AD和AD相关 痴呆（ADRD）。人类诱导多能干细胞（hiPSC）技术的进步使人类能够更好地 特异性疾病建模，特别是当鼠或啮齿动物中不存在疾病相关的遗传突变时 模型（例如，许多AD GWAS基因），但由于难以模拟体内环境而面临挑战 在当前的体外模型中。具有确定的细胞组成的微生理系统（MPS）可以提供 可扩展的，可重复的大脑模型，更好地概括了体内环境，其中临床前药物 发现努力可以转化为识别目标和化合物的更高成功率。这个项目 提出了一种使用AD的迷你脑组装类器官（类器官）微流体平台的开发 患者来源的APOE 4和同基因基因编辑的APOE 3 iPSC，以促进有效和可重复的筛选 用于AD治疗。NeuCyte采用强大的分化方案来生成神经元、星形胶质细胞和 小胶质细胞大量促进神经免疫组装体（NIA）的产生，其中3D 微环境概括了显著的离体脑表型，例如，神经退行性和细胞类型特异性 由于基因突变而导致的表型，使得能够改进可翻译的高通量临床前药物发现。 虽然同基因，但该平台也是模块化的，即，可以研究小胶质细胞突变的影响， 对神经元的影响，促进模仿人脑细胞复杂性的机制研究。的 AD/ADRD MPS微流控平台采用声学技术，能够经济地检查 通过微型化促进体外AD病理学，降低与细胞数量、试剂和药物相关的成本。 药物库的数量，以促进高通量药物筛选。第一阶段的成功完成将建立 AD/ADRD药物筛选平台商业化的可行性。

Abstract Alzheimer Disease (AD) is characterized by b-amyloid (Ab) accumulation, neurofibrillary tangles (NFTs), neuroinflammation, and widespread neuronal and synaptic loss. To date, there are no therapies available, although immunotherapies i.e., Lecanemab hold promise. Species differences underlie the difficulties in translating therapeutics uncovered in animal models for human brain-specific diseases, e.g., AD and AD-related dementia (ADRD). Human induced pluripotent stem cell (hiPSC) technological advances enable better human- specific disease modeling, particularly when disease-related genetic mutations are absent in murine or rodent models (e.g., many AD GWAS genes), but face challenges due to the difficulty in mimicking the in vivo context in current in vitro models. Microphysiological systems (MPS) with defined cellular compositions can provide scalable, reproducible brain models that better recapitulate the in vivo environment, in which preclinical drug discovery efforts can translate to a higher success rate for identified targets and compounds. This project proposes the development of a mini-brain assembled organoids (assembloids) microfluidics platform using AD patient-derived APOE4 and isogenic gene-edited APOE3 iPSCs to facilitate effective and reproducible screening for AD therapeutics. NeuCyte employs robust differentiation protocols to generate neurons, astrocytes, and microglia in large quantities facilitating the generation of NeuroImmune Assembloids (NIA) in which the 3D microenvironment recapitulates salient ex vivo brain phenotypes, e.g., neurodegenerative and cell-type specific phenotypes due to a genetic mutation, enabling improved translatable high-throughput preclinical drug discovery. While isogenic, this platform is also modular, i.e., the impact of a mutation in microglia can be studied to model effects on neurons facilitating mechanistic studies mimicking the cellular complexity of the human brain. The AD/ADRD MPS microfluidic platform incorporates acoustic technology and enables economical examination of AD pathology in vitro facilitated by miniaturization, reducing costs associated with cell numbers, reagents, and drug library quantities to facilitate high-throughput drug screening. Successful completion of Phase I will establish the feasibility for commercialization of an AD/ADRD drug screening platform.