Construction of the integrated human cortical organoids to investigate neurodevelopmental disorders

构建整合的人类皮质类器官来研究神经发育障碍

• 批准号: 10216628
• 负责人: In-Hyun Park
• 金额: $ 63.9万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-09 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10216628
• 关键词: 3-Dimensional; ASD patient; Address; Affect; Animal Model; Area; Axon; BMP7 gene; Brain; Brain imaging; Brain region; CRISPR/Cas technology; Cell Line; Cell Nucleus; Cell physiology; Cells; Cellular Structures; Chromatin Structure; Clinical; Complex; Data; Defect; Development; Diagnosis; Disease; Dorsal; Fibroblast Growth Factor; Ganglia; Generations; Genes; Genetic; Genetic Transcription; Genetic study; Genomics; Goals; Human; Human Genetics; Impairment; Insulin; Investigation; Light; Medial; Mental disorders; Methods; Molecular; Mutation; Neurobiology; Neurodevelopmental Disorder; Neurons; Organoids; Outcome; Patients; Play; Process; Prosencephalon; Proteins; Reporting; Role; SHH gene; Schizophrenia; Signal Transduction; Structure; Subthalamic structure; System; Technology; Thalamic structure; Transcriptional Regulation; WNT Signaling Pathway; Work; autism spectrum disorder; base; brain cell; brain tissue; cell type; de novo mutation; developmental disease; diencephalon; endophenotype; human pluripotent stem cell; imaging study; improved; migration; neurodevelopment; neuropsychiatric disorder; protein complex; rare variant; self-renewal; single-cell RNA sequencing; species difference; stem cells; three dimensional structure; tool; transcriptome

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.

项目摘要 诊断主要基于症状描述，很难区分 神经精神障碍，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各种神经精神疾病的遗传基础。 这些疾病表明，大多数SCZ和ASD是遗传异质性的，由罕见的遗传性疾病引起。 新生突变特别地，在SET 1/COMPASS复合蛋白或相关蛋白上的罕见突变被发现。 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ASD相关蛋白在皮质和皮质下发育中的作用将揭示其分子机制， 具有相似催化活性的蛋白质的突变导致临床上不同的病症。缺乏无障碍设施 人脑组织已经挑战了通过使用人脑组织直接解决这些问题。 因此，为了实现这一目标，我们将使用在结构和功能上都符合要求的人脑类器官， 复制发育中的人脑区域。人多能干细胞（hPSC）的使用已经被证实是一种新的干细胞。 彻底改变了人类大脑的研究hPSC经历无限的自我更新，并且可以分化成任何 细胞类型，包括脑细胞。我们已经报道了从三维（3-D）结构的生成， 重演发育中的人类皮质（hCO，皮质类器官）的hPSC，内侧神经节隆起 （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， 细胞到皮质，或相互的皮质丘脑或丘脑皮质连接。利用先进的干细胞 工具，我们将追求目标，以实现目标。1)我们将使用CRISPR/CAS9基因编辑工具来介绍 将SCZ和ASD基因突变导入hPSC细胞系，并研究其细胞和分子功能。 使用hThO和hCO进行皮质和丘脑发育。2)我们将开发方法来复制 丘脑中的多个核团，以进一步改善hThO的区域特异性。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， 由SCZ和ASD相关基因明显调节的机制，并提供前所未有的独特的 构建功能性皮质丘脑连接的平台。