RECODE-ing the Logic of Ectodermal Organoid Patterning via Combinatorial Morphogen Treatments and Longitudinal Monitoring

脑类器官是从干细胞衍生的人脑的活体模型。类器官也是了解大脑如何发育和研究神经系统疾病的有前途的工具。它们也可用于发现新药和开发治疗方法。然而，类器官难以大规模生产。该项目旨在通过创建分化地图和设计类器官当前状态的“位置跟踪器”来改善复杂脑类器官的生产。基于这些工具，将开发将分化路由到期望的类器官类型的策略。该项目中开发的方法可用于扩大STEM的参与，允许学生在类器官分化后进行远程实验。该RECODE项目的前提是，分化轨迹的定量知识与主动传感相结合，将支持脑类器官的可重复，可推广和自我校正的生产。为了系统地记录分化期间的类器官细胞类型和组成，将测量跨一系列类器官方案的单细胞中的基因表达，这通过多种类型的细胞条形码实现。这些轨迹将与体内不同大脑区域的发育进行比较。将开发由自动化和微流体技术实现的改进的生命支持系统，以支持可扩展性。将利用基于非牺牲分子和成像的方法来纵向跟踪分化期间的类器官状态，并提供缺失的营养和图案化因子。该项目的结果将支持用于疾病模型和再生医学的稳健和可重复的类器官生成的新策略。最后，通过启用远程实验，本研究中开发的方法支持将不同社区纳入科学过程。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Brain organoids are living models of the human brain derived from stem cells. Organoids are also promising tools for understanding how the brain develops and studying neurological disorders. They may also be used for discovering new drugs and developing therapies. However, organoids are difficult to produce at large scales. This project aims to improve production of complex brain organoids by creating a map of differentiation and engineering a “location tracker” of an organoid's current state. Based on these tools, strategies to route differentiation to a desired organoid type will be developed. The approaches developed in this project can be used for broadening participation in STEM, allowing student to perform remote experiments following organoid differentiation. This RECODE project is based on the premise that quantitative knowledge of differentiation trajectories combined with active sensing will support the reproducible, generalizable, and self-correcting production of brain organoids. To systematically record organoid cell types and composition during differentiation, gene expression in single cells across a range of organoid protocols will be measured, enabled by multiple types of cellular barcodes. These trajectories will be compared to the development of diverse brain regions in vivo. Improved life support systems, enabled by automation and microfluidics, will be developed to support scalability. Non-sacrificial molecular and imaging based methods will be utilized to longitudinally track organoid states during differentiation and to supply missing trophic and patterning factors. The results of this project will support novel strategies for robust and reproducible organoid generation for disease models and regenerative medicine. Finally, by enabling remote experimentation the methods developed in this study support inclusion of diverse communities in the scientific process.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.