Human Isogenic Organoid Models of Genetic Forms of Autism to Identify Convergent and Divergent Pathomechanisms in Autism

自闭症遗传形式的人类同基因类器官模型，用于识别自闭症趋同和发散的病理机制

• 批准号: 10736309
• 负责人: Stefan Aigner
• 金额: $ 79万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-07 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10736309
• 关键词: 3-Dimensional; Acceleration; Address; Affect; Architecture; Autopsy; Awareness; Biological Assay; Brain; Calcium; Categories; Cell Line; Cell model; Cells; Child; Chromium; Chromosome Mapping; Clinical; Complex; Copy Number Polymorphism; Corpus striatum structure; Data; Data Set; Defect; Development; Diagnostic; Disease model; Enzymes; Etiology; FRAP1 gene; Frequencies; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Models; Genetic Transcription; Genome; Genome engineering; Human; Image; Individual; Interneurons; Investigation; Measures; Messenger RNA; Methodology; Methods; Modeling; Molecular; Nature; Neurodevelopmental Disorder; Neurons; Noise; Operative Surgical Procedures; Organoids; Pathologic; Pathology; Pathway interactions; Patients; Phenotype; Population; Prosencephalon; Protein Biosynthesis; Proteins; RNA Editing; Recurrence; Regulation; Reproducibility; Research Personnel; Resolution; Resources; Ribosomal Interaction; Ribosomal Proteins; Ribosomes; Role; Series; Signal Transduction; Sirolimus; Site; Synapses; Testing; Tissues; Translations; Variant; apoB mRNA editing catalytic subunit; autism spectrum disorder; biomarker discovery; biomarker identification; brain tissue; candidate validation; cell type; diagnostic biomarker; disorder risk; drug testing; functional outcomes; induced pluripotent stem cell; insight; microdeletion; migration; multi-electrode arrays; neuronal circuitry; pharmacologic; predictive signature; rabies viral tracing; risk variant; single-cell RNA sequencing; stable cell line; stem cell model; stem cells; targeted treatment; therapeutic development; tool; transcriptome; transcriptomics; transgene expression; translatome

PROJECT SUMMARY Autism spectrum disorder (ASD) is a clinically complex, heterogeneous condition affecting 1 in 44 children in the U.S. The identification of common etiologies across multiple forms of genetic and idiopathic forms of ASD will critically advance diagnostic biomarker discovery and therapeutic development. Dysregulation of cellular translation has emerged as a pathophysiological mechanism common to at least a subset of ASD forms. However, systematic investigation of the cellular mechanisms that converge onto the ASD phenotype has been hampered by a paucity of robust and reproducible human ASD cellular models and scalable experimental tools for cell-type resolved characterization at the level of translation. To address these bottlenecks and to directly address the role of translational dysregulation as a common feature in ASD, we have (1) used advanced genome engineering tools to generate an extensively validated, isogenic series of induced pluripotent stem cell (iPSC) lines modeling 15 syndromic forms of ASD caused by highly penetrant gene and genome variants, representing ~10% of the total ASD population (the largest such panel created to date, to our knowledge), (2) established a robust human iPSCs-derived cortical organoid model of brain development, and (3) developed ribo-STAMP, a method for translational profiling of individual cells in heterogeneous cell populations, which is the first and only method enabling translation to be measured at single- cell resolution. In this project, we identify common and divergent pathological mechanisms in genome- engineered isogenic stem cell based organoid models of ASD, using single-cell transcriptomic and translatomic approaches. We validate our findings using cellular and functional phenotypic assays and in patient-derived iPSC models. If successful, our study will identify common and unique translation-aware single-cell resolved gene expression signatures that predict cellular and functional outcomes. We anticipate that our datasets and insights into cell-type specific deficits in gene expression of genetic forms of autism will critically accelerate the development of a unified framework that enables molecular categorization of both genetic and idiopathic cases, facilitating the identification of biomarkers and the development of targeted therapies.

项目总结 自闭症谱系障碍(ASD)是一种临床上复杂的、异质性的疾病，每44名儿童中就有1名受到影响。 美国对多种形式的遗传性和特发性ASD常见病因的识别将 关键地推进诊断生物标记物的发现和治疗开发。细胞调节失调 翻译已经成为至少一部分ASD形式共有的一种病理生理机制。 然而，对汇聚到asd表型的细胞机制进行了系统的研究。 受制于缺乏健壮和可重复的人类ASD细胞模型和可扩展的实验工具 用于翻译级别的单元格类型解析表征。为了解决这些瓶颈，并直接 解决翻译失调在ASD中的作用作为一个共同的特征，我们(1)使用高级基因组 工程工具用于产生广泛验证的等基因系列诱导多能干细胞(IPSC) 模拟由高渗透性基因和基因组变异引起的15种ASD综合征形式的线条，代表 约10%的自闭症患者(据我们所知，这是迄今为止创建的最大规模的自闭症患者小组)，(2)建立了 健壮的人类IPSCs衍生的大脑发育皮质器官模型，以及(3)开发了Ribo-Stamp，一个 异质细胞群体中单个细胞的翻译图谱方法，这是第一个也是唯一的方法 能够以单单元格分辨率测量平移的方法。在这个项目中，我们确定了共同和 基于基因组工程等基因干细胞的ASD器官模型的不同病理机制， 使用单细胞转录组和翻译组方法。我们用手机和手机验证了我们的发现 功能表型分析和患者衍生的IPSC模型。如果成功，我们的研究将确定共同的 和独特的翻译感知单细胞可分辨的基因表达特征，预测细胞和功能 结果。我们预计，我们对细胞类型特异性缺陷基因表达的数据集和洞察力 遗传形式的自闭症将关键地加速发展一个统一的框架，使分子 对遗传和特发性病例进行分类，促进识别生物标记物和 开发靶向治疗。