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增强的基于类器官的体外临床前药物 神经系统和神经退行性脑部疾病的筛查平台。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理研究的支柱。 然而，2D细胞培养物和动物不能准确地概括细胞培养物的复杂性和独特特征。 因此，人类生理学在许多关键方面与它们在体内和人类对应物表现不同 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）技术的进步，类似于 发育中的器官，称为类器官，已经产生了许多类型的器官，包括大脑 类器官这些人类类器官复制了动物中未观察到的关键器官和组织特异性特征 模型或2D细胞培养，从而提供了一个独特的机会，模拟人体器官的结构和功能 在健康和疾病的条件下。脑类器官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。具体来说，类器官是 球形，而商业微电极阵列（MEA）是平坦的，这降低了确定的准确性 神经网络组织，因为用于记录神经信号的细胞表面积有限， 类器官在平面上重塑。为了提高脑类器官在临床前研究中的价值， 疾病建模，需要一种MEA技术，能够监测尽可能多的 尽可能地保护生理上完整的类器官表面。目前还没有这样的商业平台。 该应用旨在通过利用BMSEED的可拉伸微电极来解决这个问题， 不同大小的口袋，以容纳类器官，保持其形状和生理功能，并包裹它 用微电极记录其表面的神经活动。这个新的3D平台，器官- 基于刺激和记录真空设备（OBSuRVE），集成了三个模块，（i）创建 用于类器官研究的适应性轮廓中的口袋多维电极阵列（conforMEA），（ii） 记录神经信号，和（iii）对细胞和细胞过程成像。具体而言，这项建议有三个目标。 第一个具体目标是建立OBSuRVE平台，并调整一致性多边环境协定，以满足 对类器官研究的需求。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OBSuRVE平台用于药物筛选 和疾病模型的研究。这项提案的重点是人脑类器官， 神经和神经变性疾病，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 适用于其它类型的类器官，例如，心脏球体，也用于心血管疾病模型。

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.