Development of a validated human iPSCs-derived microglia and brain organoid co-culture platform to investigate neuro-immune interactions

这个项目的首要目标是开发一个经过验证的人类小胶质细胞和皮质脑器官共培养平台，以研究神经免疫相互作用。人脑器官是从培养皿中培养的人多能干细胞(HPSCs)中提取的脑细胞的3D自组装结构。脑有机化合物模拟了人脑发育的关键方面，这使它们成为研究人类特有的脑发育和神经疾病方面的优秀模型系统。然而，脑有机化合物天生只产生外胚层起源的脑细胞，而不是来自其他血统的细胞，如小胶质细胞。小胶质细胞是存在于大脑中的免疫细胞，约占人脑细胞总数的5%-10%，在脑发育、内环境稳定和病理条件下的神经炎症中发挥着重要作用。因此，建立hPSC来源的小胶质细胞整合到3D人脑器官中的平台为研究人类小胶质细胞和建立脑部疾病模型提供了独特的机会。之前的研究表明了产生含有小胶质细胞的有机物的可行性，但未能验证该系统。在本项目中，我们旨在通过对不同条件和新方法的系统比较来优化hPSC来源的小胶质细胞和脑器官共培养方法，并通过在细胞、转录和功能水平上与新鲜的人类胚胎组织进行比较来验证平台的有效性。此外，我们将开发一条从转录数据中对神经元-免疫相互作用进行生物信息学分析的管道。

The overarching goal of this project is to develop a validated human microglia and cortical brain organoid co-culture platform to investigate neuro-immune interactions. Human brain organoids are 3D self-assembled structures of brain cells derived from human pluripotent stem cells (hPSCs) that are grown in a dish. Brain organoids mimic key aspects of human brain development, which makes them an excellent model system for studying human-specific aspects of brain development and neurological disorders. However, brain organoids innately generate only ectodermal-originated brain cells not cells from other lineages such as microglia. Microglia are resident immune cells in the brain comprising about 5-10% of total cells in the human brain and play important roles in brain development, homeostasis, and neuroinflammation during pathological conditions. Therefore, establishing a platform for hPSC-derived microglia integrated into 3D human brain organoids provides unique opportunities to study human microglia and to model brain disorders. Previous studies that have shown the feasibility of generating microglia-containing organoids failed to validate the system. In this project, we aim to optimize hPSC-derived microglia and brain organoid co-culture method by systematic comparisons of different conditions and novel approaches and validate the platform by comparing it with fresh human embryonic tissue at the cellular, transcriptomic, and functional levels. Furthermore, we will develop a pipeline for bioinformatic analyses of neuron-immune interactions from transcriptomic data.