RECODE: New technologies to illuminate and harness cadherins for the reproducible production of cortical tissue in human cerebral organoids

脑类器官是模仿人类大脑许多特征的细胞群。当人类干细胞分化成脑细胞时，这些簇就形成了。在人类大脑上进行实验面临许多挑战。因此，在许多早期大脑发育、神经系统疾病和药物发现研究中，脑类器官被用来代替大脑。脑类器官产生的高度可变性限制了这些研究的实用性。这个项目的目标是在类器官发育过程中通过控制细胞-细胞粘附来减少变异。预期的结果将是一系列细胞-细胞粘附干预措施，广泛提高脑类器官生产的可重复性，因此它们可以用作人类神经发育和疾病的可靠模型。作为该项目的一部分，研究生和本科生将在干细胞生物学和工程不同领域的团队领导方面受到指导。细胞-细胞粘附在发育过程中动态地重塑了大脑。我们的中心假设是调节细胞-细胞粘附可以从任何人类干细胞系制造可复制的脑类器官。在脑类器官分化的不同阶段，系统地询问和调节细胞粘附的新工具将被开发和实施。这将通过三个具体目标来实现：(1)开发一套技术来控制人类多能干细胞的细胞-细胞粘附；(2)明确细胞-细胞粘附调节神经元分化的机制；(3)通过新的细胞-细胞粘附调节方案测量分化后可重复性的变化。该项目的预期成果包括细胞-细胞粘附在人类神经元分化早期阶段的作用的基础知识；调节细胞-细胞粘附的新技术；培养和监测类器官的新技术；人类多能干细胞系脑类器官发育的可行分化策略。这些新的工具和见解可能会被翻译为理解在大脑类器官以外的许多细胞类型的分化中基于粘附的形态发生。该项目是由工程生物学和健康集群在英国/CBET和生物力学和机械生物学计划在英国/CMMI联合支持。该奖项反映了美国国家科学基金会的法定使命，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 clusters of cells that mimic many features of the human brain. As human stem cells differentiate into brain cells, these clusters form. Experimenting on human brains poses many challenges. As a result, brain organoids are used in place of the brain in many studies of early brain development, neurological disorders, and drug discovery research. High variability in the production of brain organoids limits the usefulness in these studies. The goal of this project is to reduce variability through control of cell-cell adhesion during organoid development. The expected outcome will be a collection of cell-cell adhesion interventions that broadly improve the reproducibility of brain organoids production so they can be used as reliable models for human neural development and disease. As part of the project, graduate and undergraduate students will be mentored in team leadership across diverse fields of stem cell biology and engineering.Cell-cell adhesion dynamically remodels the brain during development. Our central hypothesis is that modulation of cell-cell adhesion can manufacture reproducible cerebral organoids from any human stem cell line. New tools to systematically interrogate and modulate cell-cell adhesion at different stages of cerebral organoid differentiation will be developed and implemented. This will be achieved with three specific aims: (1) Develop a suite of technologies to control cell-cell adhesion in human pluripotent stem cells; (2) Identify mechanisms by which cell-cell adhesion regulates neuronal differentiation; and (3) Measure changes in reproducibility upon differentiation with new cell-cell adhesion modulation protocols. The expected outcomes of this project include fundamental knowledge of the role of cell-cell adhesion at early stages of human neuronal differentiation; new technologies to modulate cell-cell adhesion; new technologies to culture and monitor organoids; and a workable differentiation strategy for cerebral organoid development across human pluripotent stem cell lines. These new tools and insights can likely be translated to understand adhesion-based morphogenesis in the differentiation of many cell types beyond cerebral organoids.This project is being jointly supported by the Engineering Biology and Health Cluster in ENG/CBET and the Biomechanics and Mechanobiology Program in ENG/CMMI.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.