IBIS-iPSC: Organoid modeling of cortical surface area hyperexpansion in autism spectrum disorder

IBIS-iPSC：自闭症谱系障碍皮质表面积过度扩张的类器官建模

• 批准号: 10656866
• 负责人: Jason Louis Stein
• 金额: $ 86万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656866
• 关键词: Age Months; Area; Behavior; Behavior assessment; Behavioral; Biological; Biological Models; Biological Process; Blood; Brain; Brain imaging; Cell Cycle; Cell Line; Cells; Clinical; Control Groups; Development; Diagnosis; Diagnostic; Disease; Exhibits; Family; Gene Expression; Generations; Genotype; Growth; In Vitro; Individual; Individual Differences; Infant; Life; Light; Measurement; Measures; Microscopy; Modeling; Molecular; Molecular Profiling; Neocortex; Neurons; Organoids; Outcome; Participant; Pathogenesis; Pathway interactions; Peripheral Blood Mononuclear Cell; Phenotype; Play; Population; Proliferating; Reporting; Resources; Risk; Role; Sampling; Sampling Studies; School-Age Population; Siblings; Structure; Surface; Symptoms; System; Time; Tissues; Validation; Variant; autism spectrum disorder; behavior measurement; brain overgrowth; brain size; brain volume; cell behavior; cell type; cognitive ability; cohort; high risk; imaging study; in vivo; in vivo Model; individual variation; individuals with autism spectrum disorder; induced pluripotent stem cell; infancy; insight; inter-individual variation; neocortical; nerve stem cell; neurodevelopment; neurogenesis; neuroimaging; personalized medicine; phenotypic data; postnatal; proband; serial imaging; single-cell RNA sequencing; therapeutic target

Project Summary/Abstract Many individuals with monogenic and idiopathic forms of autism spectrum disorder (ASD) exhibit brain enlargement early in life. However, the underlying cellular and molecular mechanisms leading to early brain overgrowth in ASD are unknown. To identify the mechanisms leading to brain overgrowth, we will use an appropriate model system, iPSC-derived organoids, from a well-powered, deeply phenotyped cohort with multiple control groups, the Infant Brain Imaging Study (IBIS). IBIS is the largest longitudinal neuroimaging study of infants (>250 participants) at high familial risk for autism by virtue of having an older sibling/proband with ASD. Importantly, IBIS participants have previously undergone longitudinal neuroimaging at multiple time points in infancy (between 6-24 months of age) and school age, extensive behavioral assessments at these time points, as well as rare and common variant genotyping. The extensive phenotypic data generated in this cohort make it an ideal population from which to generate iPSC-derived organoid models and relate in vitro phenotypes to in vivo brain growth and behavioral trajectories. Our study also represents a unique opportunity to evaluate how well organoid phenotypes model the in vivo brain growth trajectories of the individual from whom they were derived. In this proposal, we will derive and validate iPSCs from blood for participants from high risk families who developed ASD (HR+), high risk participants who did not develop ASD (HR-), and low risk individuals without ASD (LR-) totaling 99 participants. We will differentiate the iPSC lines to cortical organoids to model inter- individual differences in brain development. We will use single cell (sc)RNA-seq to identify cell types, cell cycle states, and differentiation trajectories in each participant-derived organoid across two time points modeling the period of cortical neurogenesis, totaling 2.38M sequenced cells. We will validate cell type counts and states using tissue clearing followed by lightsheet microscopy of the cortical organoids. We will identify cell types, fate decisions, and cell cycle states that correlate with both cross-sectional and longitudinal cortical surface area growth and ASD symptoms and cognitive ability over time. Leveraging this unique, deeply characterized clinical cohort, we will determine both the in vivo relevance of cortical organoids and the cellular and molecular mechanisms underlying brain overgrowth in ASD.

项目摘要/摘要 许多具有自闭症谱系障碍（ASD）的单基因和特发性形式的人表现出大脑 生命早期扩大。但是，导致早期大脑的潜在细胞和分子机制 ASD中的过度生长是未知的。为了确定导致大脑过度生长的机制，我们将使用 适当的模型系统，IPSC衍生的类器官，来自富有驱动的，深厚的表型队列 多个对照组，婴儿脑成像研究（IBIS）。 IBIS是最大的纵向神经成像研究 凭借与ASD的较旧的兄弟姐妹/概率，婴儿（> 250名参与者）对自闭症的家庭风险很高。 重要的是，IBIS参与者以前在多个时间点进行了纵向神经影像学。 婴儿期（在6-24个月之间）和学龄，在这些时间点进行广泛的行为评估， 以及稀有和常见的变体基因分型。该队列中产生的广泛表型数据使得 从中产生IPSC衍生的类器官模型并将体外表型与IN相关联的理想人群 体内大脑的生长和行为轨迹。我们的研究还代表了评估如何评估的独特机会 良好的器官表型模拟了他们所在的个体体内脑生长轨迹 衍生的。在此提案中，我们将为来自高风险家庭的参与者而从血液中得出并验证IPSC 发达的ASD（HR+），未发展ASD（HR-）的高风险参与者，而没有人为低风险的人 ASD（LR-）总计99名参与者。我们将将IPSC系与皮质器官区分开，以建模 大脑发育中的个体差异。我们将使用单个单元格（SC）RNA-Seq识别细胞类型，细胞周期 在两个时间点上，每个参与者衍生的类器官中的状态和分化轨迹建模 皮质神经发生的周期，总计238万个测序细胞。我们将验证细胞类型计数和状态 使用组织清除，然后使用皮质器官的灯表显微镜。我们将确定细胞类型，命运 决策，细胞周期状态与横截面和纵向皮质表面积相关 随着时间的流逝，生长和ASD症状和认知能力。利用这种独特的，深刻的临床特征 队列，我们​​将确定皮质器官的体内相关性以及细胞和分子 ASD中大脑过度生长的机制。