A physiologically relevant pre-clinical drug screening platform for Alzheimer's Disease and Traumatic Brain Injury with integrated stretchable microelectrodes

摘要 拟议的工作旨在商业化的微流控芯片为基础的平台建模 阿尔茨海默病和阿尔茨海默病相关痴呆（AD/ADRD）的临床前体外机制 研究和药物测试。AD/ADRD是一个日益增长的健康问题，占所有痴呆症的50-75% 病例，目前影响估计580万美国人。除了人类的痛苦， AD/ADRD的年成本为2900亿美元。目前FDA批准的治疗方法只能帮助控制 这种疾病然而，尽管有数百种临床试验，但没有治疗方法可以阻止或逆转其进展。 审判成功开发治疗的一个主要障碍是缺乏合适的临床前模型。在 此外，有大量的流行病学证据表明，创伤性脑损伤与 损伤（TBI）、长期AD/ADRD病理和认知下降。然而，AD和 TBI未知。本申请旨在通过开发一种新型的微流体技术来解决这两个问题， 基于三维体外AD模型，并将该芯片与BMSEED现有的体外TBI模型平台相结合， 微电极阵列拉伸刺激和记录设备（MEASSuRE），以满足以下需求 临床前AD/ADRD研究。这个新平台提供了一个有效的和生理相关的临床前 用于AD治疗的药物筛选平台。该平台也非常适合调查TBI的影响 有或没有预先存在的遗传倾向发展AD的人。关键的创新在于 用于微流体药物中神经元健康的功能评估的可拉伸微电极阵列 筛选平台，以及调查AD和TBI之间的机制联系和相似性的能力 在3D细胞培养基质（3D-sMEA）中使用该可拉伸微电极阵列。3D-MEASSURE 平台提供了一个更真实的在体外复制的天然在体内的生化和生物力学 与现有的2D系统相比，细胞的微环境。第一阶段的重点是证明 Concept（概念）使用单孔（SW）3D-MEASSuRE平台，细胞来源于遗传修饰的 (3xTg，5xFAD）和野生型小鼠。第二阶段的目标是：（a）通过发展高效率的 通量多孔（MW）3D-MEASSuRE平台，和（B）通过以下方式增加与临床翻译的相关性 使用源自AD的诱导多能干细胞（hiPSC）的人细胞评估平台 患者和年龄匹配的对照组。3D-MEASSuRE平台用于遗传学研究的能力 易患AD和大脑中不同细胞类型之间的串扰在介导 将评估和验证TBI相关应变损伤后的神经元健康。本提案的重点是 开发AD/ADRD的临床前药物筛选平台，然而， 研究也将适用于其他神经退行性疾病的药物筛选，例如，帕金森 疾病在第二阶段结束时，3D-MEASSuRE平台将为市场做好准备。

Abstract The proposed work aims at the commercialization of a microfluidic chip-based platform for modeling Alzheimer’s disease and Alzheimer’s disease related dementias (AD/ADRD) for preclinical in vitro mechanistic studies and drug testing. AD/ADRD are a growing health concern, accounting for 50-75% of all dementia cases, and currently affecting an estimated 5.8 million Americans. In addition to the human suffering, the annual cost of AD/ADRD is $290 billion. Current FDA approved treatments only help manage the symptoms of the disease. However, there is no treatment to stop or reverse its progression despite hundreds of clinical trials. A major obstacle to successful treatment development is the dearth of suitable preclinical models. In addition, there is substantial epidemiological evidence of an intricate relationship between traumatic brain injury (TBI), longer-term AD/ADRD pathology, and cognitive decline. However, the exact link between AD and TBI is not known. This application aims to solve both of these problems by developing a novel microfluidics- based 3D in vitro AD model, and merging this chip with BMSEED’s existing in vitro TBI model platform, the MicroElectrode Array Stretching Stimulating und Recording Equipment (MEASSuRE), to meet the needs for pre-clinical AD/ADRD research. This new platform presents an efficient and physiologically relevant pre-clinical drug screening platform for AD treatments. The platform is also well-suited to investigate the effects of a TBI on a person with or without a pre-existing genetic disposition to develop AD. The key innovations are the use of a stretchable microelectrode array for functional assessment of neuronal health in a microfluidics drug screening platform, and the capability to investigate the mechanistic links and similarities between AD and TBI using this stretchable microelectrode array in a 3D cell culture matrix (3D-sMEA). This 3D-MEASSuRE platform provides a more realistic in vitro facsimile of the natural in vivo biochemical and biomechanical microenvironment of the cells compared to existing 2D systems. Phase I is focused on demonstrating Proof-of- Concept (PoC) using a single-well (SW) 3D-MEASSuRE platform with cells derived from genetically modified (3xTg, 5xFAD) and wild type mice. Phase II is directed towards (a) improving efficiency by developing a high throughput multi-well (MW) 3D-MEASSuRE platform, and (b) increasing relevance to clinical translation by evaluating the platform using human cells derived from induced-pluripotent stem cells (hiPSCs) from AD patients and age matched controls. The capability of the 3D-MEASSuRE platform for research on the genetic pre-disposition to develop AD and the role of crosstalk between different cell types in the brain in mediating neuronal health after TBI-relevant strain injury will be evaluated and validated. The focus of this proposal is the development of a pre-clinical drug screening platform for AD/ADRD, however, the products developed in this research will also be applicable in drug screening for other neurodegenerative diseases, e.g., Parkinson’s Disease. At the end of Phase II, the 3D-MEASSuRE platform will be ready for the marketplace.