Biological substrates of risk and resilience using patient-derived stem cells

使用患者来源干细胞的风险和复原力的生物基质

基本信息

批准号：
10240561
负责人：
FLORA M VACCARINO
金额：
$ 31.73万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-07 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10240561
关键词：
3-Dimensional Affect Attention Biological Biology Brain Brain imaging Cell Line Cell model Cellular Structures Cerebral cortex Child Clinical Collaborations Copy Number Polymorphism Data Deltastab Development Diagnosis Discipline Disease Early Diagnosis Fathers Fetal Development Functional Imaging Funding Mechanisms Future Gene Expression Gene Expression Profile Gene Expression Profiling Genes Genetic Genetic Risk Human Image In Vitro Inhibitory Synapse Link Macrocephaly Magnetic Resonance Imaging Measures Modeling Molecular Mutation Neurobiology Neurons Organoids Pathway Analysis Patient Recruitments Patients Phenotype Production Rest Risk Risk Factors Severities Siblings Structure Symptoms Synapses Syndrome Work autism spectrum disorder autistic children base cellular imaging clinical predictors cohort connectome density differential expression disorder risk early detection biomarkers excitatory neuron exome sequencing fetal gray matter high resolution imaging high risk imaging approach in vitro Model induced pluripotent stem cell inhibitory neuron male molecular imaging neurodevelopment predict clinical outcome predictive modeling prenatal prenatal disorder proband protective factors resilience stem cells synaptogenesis tool transcriptome transcriptome sequencing transcriptomics variant detection white matter

项目摘要

Autism spectrum disorder (ASD) is a disorder of prenatal brain development. While syndromic forms of ASD have received considerable attention, to what extent findings in these heterogeneous disorders apply also to the broader or idiopathic form of ASD with no identified single genetic risk is unclear. In this project, we will study how the normal trajectory of prenatal neurobiological development of the brain is disrupted in idiopathic ASD. To identify neurobiological factors that are associated with risk or protection from ASD during prenatal development, we will recruit participants from a well-characterized cohort of younger siblings of children with ASD, who were followed longitudinally. The siblings will be either concordant for ASD diagnosis (ASD:ASD; n=12 pairs) or will be discordant (ASD:TYP; n=12 pairs). We will use induced pluripotent stem cells (iPSC) derived cortical organoids, 3D cellular structures which model in vitro the fetal development of the human cerebral cortex. Organoids will be analyzed by high resolution imaging approaches, molecular tools and transcriptomics. In Aim 1 we will obtain sets of biological measures (excitatory/inhibitory neuron fate, density of synapse, and neuronal arborization), comparing and contrasting phenotypes between ASD:ASD concordant sibs ASD:TYP discordant sibs. This comparison will refine our ability to isolate risk/protective factors that will be exclusively at work in the discordant pairs. In Aim 2 we will perform global gene expression analysis by RNA-seq and network analyses, aiming at finding differences in gene expression and network organization between the ASD:ASD concordant network and the ASD:TYP discordant network. We will perform correlation analyses where neurobiological measures and gene expression will be correlated with each other and with clinical severity scores. The correlations between neurobiological and gene expression measures with symptoms severity may help discriminate between risk and protection. In Aim 3, in collaboration with Project 2, we will obtain structural MRI and BOLD-based functional connectivity data on the concordant (ASD:ASD) and discordant (ASD:TYP) sib pairs that participate in Aim 1 and Aim 2 studies. We will then make correlations between imaging and neurobiological and gene expression measures. We hypothesize that increased inhibitory neuron fate in ASD may be correlated with less efficient cortical network connectivity and that increased synaptogenesis and neuronal arborization may be correlated with higher gray matter ratio, and also to altered connectivity. This project will feed data to the statistical core, where imaging and neurobiological measures can be used to predict clinical severity, allowing a more powerful analysis of risk factors for ASD. In summary, our in vitro ASD risk human cellular model will allow, in principle, to develop future biomarkers for early diagnosis and the exploration of new treatment options based on the underlying biology.

自闭症谱系障碍 (ASD) 是一种产前大脑发育障碍，也是 ASD 的综合症形式。这些异质性疾病的发现在多大程度上也适用于没有确定的单一遗传风险的更广泛或特发性 ASD 形式尚不清楚。研究特发性产前大脑神经生物学发育的正常轨迹是如何被破坏的 ASD。确定与产前 ASD 风险或保护相关的神经生物学因素。为了发展，我们将从一群具有良好特征的患有以下疾病的儿童的弟弟妹妹中招募参与者 ASD，纵向随访的兄弟姐妹的 ASD 诊断将一致（ASD：ASD； n=12 对）或不一致（ASD:TYP；n=12 对）。我们将使用诱导多能干细胞 (iPSC)。衍生的皮质类器官、3D 细胞结构，可在体外模拟人类胎儿的发育大脑皮层将通过高分辨率成像方法、分子工具和在目标 1 中，我们将获得一组生物学测量值（兴奋性/抑制性神经元命运、神经元密度）。突触和神经元树枝化），比较和对比 ASD:ASD 一致性之间的表型同胞 ASD：TYP 不一致的同胞这种比较将提高我们隔离风险/保护因素的能力。在目标 2 中，我们将通过以下方式进行全局基因表达分析。 RNA-seq和网络分析，旨在发现基因表达和网络组织的差异我们将在 ASD:ASD 一致网络和 ASD:TYP 不一致网络之间进行关联。分析神经生物学测量和基因表达之间的相互关系以及与临床严重程度评分与神经生物学和基因表达测量之间的相关性。症状严重程度可能有助于区分风险和保护。在目标 3 中，与项目 2 合作，我们将获得一致词 (ASD:ASD) 的结构 MRI 和基于 BOLD 的功能连接数据然后，我们将对参与目标 1 和目标 2 研究的不一致 (ASD:TYP) 同胞对进行关联。成像、神经生物学和基因表达测量之间的关系。自闭症谱系障碍（ASD）中的抑制性神经元命运可能与皮质网络连接效率较低有关，并且突触发生和神经元树枝化的增加可能与较高的灰质比例相关，并且该项目将向统计核心提供数据，其中包括成像和神经生物学。测量可用于预测临床严重程度，从而对自闭症谱系障碍的危险因素进行更强有力的分析。总之，我们的体外 ASD 风险人类细胞模型原则上将允许开发未来的生物标志物基于基础生物学的早期诊断和新治疗方案的探索。