Relating interindividual differences in cerebral organoid models to longitudinal infant brain growth and cognitive development

项目摘要/摘要 诱导多能干细胞(IPSCs)和人脑有机体有望为我们提供关于 神经发育障碍患者的脑发育变化。尽管分子的保真度 皮质类有机化合物已经相对于初级胎儿大脑参考数据集进行了评估，目前尚不清楚如何进行 皮质器官表型可以预测出生后脑纵向发育的个体差异。 确定哪些体外表型与体内性状有关将有助于解释实验 实施有机化合物以了解神经发育障碍的机制。为了测试这种关系，它 是从特征良好的个体衍生出IPSC系的群体所必需的。本提案使用IPSC- 从婴儿脑成像研究(IBIS)参与者那里提取脑器官来测试两者之间的关系 有机类神经上皮芽结构和基因表达表型与活体脑发育的关系 和认知行为测量。具体地说，该提案使用从5个IPSC行生成的17个IPSC行 参与者没有神经精神障碍家族史，9名有自闭症兄弟姐妹的参与者， 以及3名患有自闭症的参与者，每人在6个月、12个月和/或24个月大时至少接受2次脑MRI扫描。全 参与者有马伦斯早期学习量表、Vinland适应行为和自闭症诊断 24月龄观察量表评分。每株IPSC克隆3个。要确定内部- 我们将评估IPSC捐赠者之间以及来自同一个人的IPSC克隆内的个体差异 单细胞基因的表达和器官结构在分化过程中的变化。单细胞RNA测序将 在神经上皮芽形成(分化后14天)和神经元成熟(84天)后进行 后分化)。完整的有机免疫标记将在这些时间点和更早的时间完成 神经发生(分化后56天)。完成后，该项目将确定优势和 大脑器官作为人类皮质发育的可重复性、预测性模型的局限性。这 Proposal寻求利用以下网址提供的严谨课程、指导、网络和技术培训 北卡罗来纳大学教堂山分校和IBIS的合作者。通过完成这项建议， 申请者将拥有适当的技术培训和沟通技能，以成为一名独立的 在神经发育领域拥有强大的专业网络的调查员。

PROJECT SUMMARY/ABSTRACT Induced pluripotent stem cells (iPSCs) and human cerebral organoids are poised to provide insight into how brain development changes in individuals with neurodevelopmental disorders. Though the molecular fidelity of cortical organoids have been assessed relative to the primary fetal brain reference datasets, it is unknown how well cortical organoid phenotypes predict inter-individual differences in postnatal longitudinal brain growth. Identifying what in vitro phenotypes are related to in vivo traits will be useful for interpreting experiments implementing organoids to understand mechanisms of neurodevelopment disorders. To test this relationship, it is necessary to derive a population of iPSC lines from well-characterized individuals. This proposal uses iPSC- derived cerebral organoids from participants in the Infant Brain Imaging Study (IBIS) to test the relationship between organoid neuroepithelial bud structure and gene expression phenotypes to in vivo brain development and cognitive behavioral measurements. Specifically, this proposal uses 17 iPSC lines generated from 5 participants without a family history of neuropsychiatric disorders, 9 participants who had a sibling with ASD, and 3 participants with ASD, each with at least 2 brain MRI scans at 6, 12 and/or 24 months of age. All participants have Mullens Scale of Early Learning, Vineland Adaptive Behavior and Autism Diagnostic Observation Scale score at 24 months of age. 3 clones of each iPSC line were generated. To identify inter- individual differences between iPSC donors and within iPSC clones from the same individual, we will assess single cell gene expression and organoid structure across the differentiation. Single cell RNA sequencing will be performed after neuroepithelial buds form (14 days post differentiation) and neuronal maturation (84 days post differentiation). Intact organoid immunolabeling will be completed at these time points and earlier in neurogenesis (56 days post differentiation). When completed, this project will identify the strengths and limitations of cerebral organoids as reproducible, predictive models of human cortical development. This proposal seeks to leverage the rigorous coursework, mentorships, networks and technical training available at the University of North Carolina at Chapel Hill and through IBIS collaborators. By completing this proposal, the applicant will have the appropriate technical training and communication skills to become an independent investigator with a strong professional network in the field of neurodevelopment.