The first adaptable, 3D-formfitting microelectrode array for organoid-based models of neurological and neurodegenerative diseases

摘要 拟议的工作旨在开发一种基于有机化合物的增强型体外临床前药物。 神经和神经退行性脑疾病筛查平台。2D体外细胞培养和非 几十年来，人类动物一直是临床前药物开发和机制研究的中流砥柱。 然而，2D细胞培养和动物并不能准确地概括 因此，人类生理学在许多关键方面都不同于它们在体内和人类的同行 细胞行为的特征，限制了我们准确模拟大脑疾病的能力。感谢 人类诱导多能干细胞(HiPSC)技术进展，复杂结构类似于 发育中的器官，称为类器官，已经为许多类型的器官产生，包括大脑。 有机化合物。这些人类有机化合物复制了在动物身上没有观察到的关键器官和组织特有的特征 模型或2D细胞培养，从而为模拟人体器官结构和功能提供了独特的机会 在健康和疾病条件下。脑有机体研究的一个主要限制是缺乏足够的 监测神经网络的空间和时间组织的仪器。具体地说，有机化合物是 球形而商用微电极阵列(MEA)是扁平的，这降低了测定的准确性 神经网络组织，因为用于记录神经信号的细胞表面积是有限的 有机物体在平面上进行重塑。提高脑有机化合物在临床前研究和研究中的价值 疾病建模，需要一种MEA技术，能够监控多个 尽可能保持生理上完好无损的器官表面。目前还不存在这样的商业平台。 这项应用旨在通过利用BMSEED的可伸展微电极来解决这一问题 不同大小的口袋，以容纳类器官，保持其形状和生理功能，并将其包裹 用微电极记录其表面的神经活动。这个新奇的3D平台，有机生物- 基于刺激和记录真空设备(OBSuRVE)，集成了三个模块：(I)创建 用于有机物研究的自适应轮廓中的口袋多维电极阵列(ConforMEA)，(Ii) 记录神经信号，以及(Iii)成像细胞和细胞过程。具体地说，这项提议有三个目标。 第一个具体目标是构建OBSuRVE平台，并调整ConforMEA以满足 需要有机化合物的研究。第二个具体目标是评估电气和机械 平台的属性。第三个具体目标是验证用于药物筛选的OBSuRVE平台 以及使用脑器官进行疾病建模。这项提议的重点是人脑有机化合物，因为 神经和神经退行性疾病，如自闭症、阿尔茨海默病和帕金森病， 是我们社会面临的最普遍和代价最高的健康问题之一。然而，结果将是 也适用于心血管疾病模型的其他类型的有机体，例如心脏球体。

Abstract The proposed work aims at the development of an enhanced organoid-based in vitro pre-clinical drug screening platform for neurological and neurodegenerative brain diseases. 2D in vitro cell cultures and non- human animals have been the mainstay of pre-clinical drug development and mechanistic studies for decades. However, 2D cell cultures and animals do not accurately recapitulate the complexity and unique features of human physiology, thus behave differently from their in vivo and human counterparts in many key characteristics of cellular behavior, limiting our ability to accurately model brain diseases. Thanks to advancements in human induced pluripotent stem cell (hiPSC) technology, complex structures resembling developing organs, named organoids, have been generated for many types of organs, including brain organoids. These human organoids replicate critical organ and tissue-specific features not observed in animal models or 2D cell cultures, thus providing a unique opportunity to model human organ structure and function under healthy and disease conditions. A major limitation for brain organoid research is the lack of adequate instrumentation to monitor spatial and temporal organization of neural networks. Specifically, organoids are spherical whereas commercial microelectrode arrays (MEAs) are flat, which reduces the accuracy to determine neural network organization because the cellular surface area for recording neural signals is limited and organoids remodel on flat surfaces. To enhance the value of brain organoids for preclinical research and disease modeling, an MEA technology is needed that enables monitoring of neural signals across as much of the surface of the physiologically intact organoid as possible. No such commercial platform currently exists. This application aims to solve this problem by utilizing BMSEED’s stretchable microelectrodes to create pockets of variable sizes to contain the organoid, retain its shape and physiological function, and envelope it with microelectrodes for recording of neural activity across its surface. This novel 3D platform, the Organoid- Based Stimulating und Recording Vacuum Equipment (OBSuRVE), integrates three modules that (i) create the pockets in the adaptable contour for organoid research Multidimensional Electrode Array (conforMEA), (ii) record neural signals, and (iii) image cells and cellular processes. Specifically, this proposal has three aims. The first specific aim is focused on building the OBSuRVE platform, and to adapt the conforMEAs to meet the need for organoids research. The second specific aim is the evaluation of the electrical and mechanical properties of the platform. The third specific aim is the validation of the OBSuRVE platform for drug screening and disease modeling using brain organoids. The focus of this proposal are human brain organoids because neurological and neurodegenerative diseases, such as Autism, Alzheimer’s Disease, and Parkinson’s Disease, are among the most prevalent and costly health problems facing our society. However, the results will be applicable to other types of organoids, e.g., cardiac spheroids, for cardiovascular disease models as well.