Construction of the integrated human cortical organoids to investigate neurodevelopmental disorders

项目概要 主要根据症状描述进行诊断，很难区分其潜在机制 神经精神疾病，包括精神分裂症（SCZ）和自闭症谱系障碍（ASD）。最近 先进的人类遗传学研究已经确定了多种神经精神疾病的遗传基础 疾病，表明大多数 SCZ 和 ASD 具有遗传异质性，并且是由罕见的疾病引起的 新突变。特别是 SET1/COMPASS 复合蛋白或相关蛋白的罕见突变 在 SCZ 或 ASD 患者中明显发现。我们的中心假设是研究 SCZ 的功能 或皮层和皮层下发育中的 ASD 相关蛋白将揭示分子机制 具有相似催化活性的蛋白质突变会导致临床上不同的疾病。缺乏可访问性 利用人脑组织直接解决这些问题一直是人类脑组织面临的挑战。 因此，为了实现这一目标，我们将使用在结构和功能上 再现正在发育的人类大脑区域。人类多能干细胞 (hPSC) 的使用 彻底改变了人脑研究。 hPSC 可以进行无限的自我更新，并且可以分化成任何类型的细胞。 细胞类型，包括脑细胞。我们已经报道了 3 维 (3-D) 结构的生成 概括发育中的人类皮质（hCO、皮质类器官）、内侧神经节隆起的 hPSC （hMGEO，MGE 类器官），或间脑丘脑（hThO，丘脑类器官）。将 hCO 与 hMGEO 融合，或 hThO 再现了发育上不同的两个区域的相互作用，例如 MGE 的切向迁移 细胞与皮质，或皮质丘脑或丘脑皮质的相互连接。使用先进的干细胞 工具，我们将追求目标以实现目标。 1）我们将使用CRISPR/CAS9基因编辑工具来引入 将 SCZ 和 ASD 基因突变入 hPSC 系，并研究其细胞和分子功能 使用 hThOs 和 hCOs 进行皮质和丘脑发育。 2）我们将开发重现的方法 丘脑中的多个核，以进一步提高 hThOs 的区域规范。 3）我们将调查 SCZ 和 ASD 基因在皮质和皮质下连接中的功能。总的来说，我们先进的人类大脑 基于类器官的方法与基因组学和神经生物学工具相结合将定义分子 机制明显受 SCZ 和 ASD 相关基因调控，并提供前所未有的独特 构建功能性皮质丘脑连接的平台。

Project Summary Diagnosed mainly based on symptomatic description, it is difficult to distinguish the underlying mechanisms of neuropsychiatric disorders, including schizophrenia (SCZ) and autism spectrum disorders (ASD). Recent advanced human genetic studies have identified the genetic underpinning of the a variety of neuropsychiatric disorders, showing that majority of SCZ and ASD are genetically heterogeneous and caused by with rare de novo mutations. Particularly, a rare mutations at SET1/COMPASS complex proteins or related proteins were distinctly discovered at SCZ or ASD patients. Our central hypothesis is that investigating the function of SCZ- or ASD-associated proteins in cortical and subcortical development will reveal the molecular mechanism how mutations of proteins with similar catalytic activity result in clinically distinct disorders. The paucity of accessible human brain tissues has been challenging to directly addressing these questions by using human brain tissue. Thus, in order to achieve the goal, we will use human brain organoids that structually and functionally reproduce the developing human brain regions. The use of human pluripotent stem cells (hPSCs) has revolutionized the human brain studies. hPSCs undergo unlimited self-renewal and can differentiate into any cell types, including brain cells. We have reported the generation of 3-dimensional (3-D) structures from hPSCs that recapitulate the developing human cortex (hCO, cortical organoids), medial ganglionic eminence (hMGEO, MGE organoid), or diencephalic thalamus (hThO, thalamic organoids). Fusing hCO with hMGEO, or hThO reproduced the interaction of developmentally distinct two regions, such as tangential migration of MGE cells to cortex, or reciprocal corticothalamic or thalamocortical connections. Using the advanced stem cell tools, we will purse the aims to achieve the goal. 1) We will use CRISPR/CAS9 gene editing tools to introduce mutations of SCZ and ASD genes into hPSC lines, and investigate the cellular and molecular function in cortical and thalamic development using hThOs and hCOs. 2) We will develop methods to reproduce the multiple nuclei in thalamus to further improve the regional specification of hThOs. 3) We will investigate the function of SCZ and ASD genes in cortical and subcortical connectivity. Overall, our advanced human brain organoid-based approaches combined with genomics and neurobiological tools will define the molecular mechanism distinctly regulated by SCZ and ASD-associated genes and provide unprecedented unique platforms to construct the functional corticothalamic connection.