Advancing the functional maturity of brain organoids by synthetic afferentation.

项目总结 人类多能干细胞(HiPSCs)是理解支配 人类大脑的发育和神经系统疾病的建模。最近的一个重要事件 这项技术的应用是三维(3D)脑细胞培养或“有机类”的产生。脑区 直到中晚期，有机化合物已经被证明在结构、转录和功能上具有相似性。 与早产儿人类脑电记录和人类转录组图谱比较的妊娠期人脑。 目前，它们代表了目前可用的最接近天然人脑组织的细胞模型。当一个强大的 探索产前发育机制的系统试图促进大脑的成熟状态 有机化合物已被证明是递增的，不确定的，或者复制得很差。因此，大脑器官系统 仍然在很大程度上不适合对出生后的大脑进行建模。非常有必要开发一种新的 产生适合于审问发育后期的脑器官模型。这 提案的重点是理解生理输入作为更健壮的、 脑有机体活动的可重复的、成熟的生理状态。该项目将开发方法 用于引入与发育相关的输入，并读出人脑有机体中的神经元活动。 来自其他系统的广泛工作已经确定，传入网络活动用于建立、维护 并完善活跃的功能大脑回路。在这里，我们将测试一系列替代或模仿的创新策略 通过对人脑器官的长时间图案化刺激而错过外部输入。然后我们将观察到 从而对网络活动和基因表达产生影响。我们将把这些观察结果作为现有数据的基准 SET来自人类皮质器官以及人类和啮齿类动物的大脑。已建立的工具和方法 这里旨在很容易地适应其他器官系统的细胞培养模型，对于这些器官系统，神经元输入 是很重要的。如果成功，这个项目将建立一个新的平台，用于审问依赖活动的人 并将使人类大脑发育和人类进入更高级的阶段 神经学和神经精神病学疾病状态。

PROJECT SUMMARY Human pluripotent stem cells (hiPSCs) are a powerful tool for understanding the principles that govern the development of the human brain and for modeling diseases of the nervous system. One important recent application of this technology is the generation of tri-dimensional (3D) brain cell cultures or "organoids." Brain organoids have been shown to develop structural, transcriptional, and functional similarities up to the mid-to-late gestation human brain as compared to preterm human EEG recordings and human transcriptome profiles. Currently they represent the closest cellular model to native human brain tissue available. While a powerful system for probing mechanisms of prenatal development attempts to advance the maturation state of brain organoids have proved to be incremental, inconclusive, or poorly reproduced. As a result, brain organoid systems remain largely inappropriate for modeling the postnatal brain. There is a significant need to develop a new generation of brain organoid models that are appropriate for interrogating later developmental stages. This proposal focuses on understanding the role of physiological inputs as a necessary driver of more robust, reproducible, and mature physiological states of activity in brain organoids. This project will develop approaches for introducing developmentally-relevant inputs and for reading out neuronal activity in human brain organoids. Extensive work from other systems has established that afferent network activity serves to establish, maintain, and refine active functional brain circuits. Here, we will test a series of innovative strategies to replace or mimic missing external inputs via prolonged patterned stimulation of human brain organoids. We will then observe the resulting effects on network activity and gene expression. We will benchmark these observations to existing data sets from human cortical organoids as well as the human and rodent brain. The tools and approaches established here are intended to be readily adapted to cell culture models of other organ systems for which neuronal inputs are important. If successful, this project will establish a novel platform for interrogating activity-dependent maturation and will enable access to more advanced stages of human brain development and human neurological and neuropsychiatric disease states.