Modeling Inherited Neurodevelopmental Disorders with Human Induced Pluripotent Stem Cells

用人类诱导多能干细胞模拟遗传性神经发育障碍

• 批准号: 10397399
• 负责人: HYDER A JINNAH
• 金额: $ 58.65万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10397399
• 关键词: Address; Animal Model; Autopsy; Biological Process; Biology; Brain; CRISPR/Cas technology; Cell Line; Cell Lineage; Cell model; Cells; Clinical; Complex; Defect; Development; Disease; Dissection; Environmental Risk Factor; Enzymes; Epigenetic Process; Evaluation; Event; Experimental Models; Fibroblasts; Future; Gene Expression Profiling; Genes; Genetic; Genotype; HPRT1 gene; Human; Hypoxanthine Phosphoribosyltransferase; Individual; Inherited; Lesch-Nyhan Syndrome; Link; Mendelian disorder; Methods; Modeling; Molecular; Mutation; Neurobiology; Neurodevelopmental Disorder; Neuronal Dysfunction; Neurons; Pathogenesis; Pathway interactions; Patients; Penetrance; Pharmacology; Phenotype; Poverty; Purines; Resources; Role; Severities; Severity of illness; Source; Time; Variant; base; brain dysfunction; cell type; clinically relevant; developmental disease; dopaminergic neuron; gene therapy; imaging study; induced pluripotent stem cell; insight; neurite growth; neurobehavioral; neuron development; novel; patient variability; prototype; stem cell model; tool; transcriptome sequencing

PROJECT SUMMARY The pathogenesis of neurodevelopmental disorders is challenging to address for many reasons. The human brain is relatively inaccessible for direct evaluation, human autopsy and imaging studies provide limited opportunities to study molecular or cellular mechanisms, animal models sometimes do not replicate important disease features, and immortalized neuron-like cell lines cannot be used to model normal development. Induced pluripotent stem cells (iPSCs) provide a powerful new experimental tool to address these limitations. They can be created from patients with known developmental disorders caused by defined mutations, they provide a renewable resource, and they can be differentiated into specific cell lineages to provide species-specific and lineage-specific experimental models to study developmental mechanisms at the cellular and molecular levels. In the current proposal we address several fundamental questions that are broadly relevant to iPSC modeling for neurodevelopmental disorders. We focus on Lesch-Nyhan disease (LND) as a prototype disorder, because it has numerous features that make it an unusually tractable Mendelian disorder for iPSC modeling. LND and its milder variants are caused by mutations in the HPRT1 gene, resulting in deficiency of the purine salvage enzyme, hypoxanthine-guanine phosphoribosyltransferase (HGprt). The neurobehavioral abnormalities in this disorder have been linked with developmental dysfunction of brain dopamine neurons. Aim 1 focuses on establishing a well-characterized bank of iPSCs across the spectrum of disease severity, to address issues related to how specific mutations causing different disease severity are reflected in iPSC models. Aim 2 focuses on establishing a well-characterized bank of isogenic iPSCs in which specific mutations have been introduced via gene editing methods, to address issues related to the genetic background of individuals from which iPSC models are derived. In Aim 3 we differentiate these lines into dopamine neurons, to address how iPSC models can reveal the timing and mechanisms of pathogenesis related to LND. Overall, this project will address fundamental questions that are broadly relevant for iPSC modeling of neurodevelopmental disorders, as well as more specific insights into the pathogenesis of the neurobehavioral abnormalities of LND.

项目概要 由于多种原因，神经发育障碍的发病机制很难解决。人类 大脑相对难以直接评估，人体尸检和影像学研究提供的信息有限 研究分子或细胞机制的机会，动物模型有时无法复制重要 疾病特征和永生化神经元样细胞系不能用于模拟正常发育。诱发 多能干细胞（iPSC）提供了一种强大的新实验工具来解决这些局限性。他们可以 是由患有已知由特定突变引起的发育障碍的患者创建的，它们提供了 可再生资源，并且它们可以分化成特定的细胞谱系，以提供物种特异性和 用于研究细胞和分子水平发育机制的谱系特异性实验模型。 在当前的提案中，我们解决了与 iPSC 建模广泛相关的几个基本问​​题 用于神经发育障碍。我们重点关注 Lesch-Nyhan 病 (LND) 作为一种原型疾病，因为 它具有众多特征，使其成为 iPSC 建模中异常容易处理的孟德尔疾病。 LND 和 其较温和的变异是由 HPRT1 基因突变引起的，导致嘌呤补救不足 酶，次黄嘌呤鸟嘌呤磷酸核糖转移酶（HGprt）。此过程中的神经行为异常 疾病与大脑多巴胺神经元的发育功能障碍有关。目标 1 侧重于 建立一个涵盖各种疾病严重程度的特征良好的 iPSC 库，以解决问题 与导致不同疾病严重程度的特定突变如何反映在 iPSC 模型中有关。目标2重点 建立一个特征良好的同基因 iPSC 库，其中引入了特定突变 通过基因编辑方法，解决与 iPSC 个体遗传背景相关的问题 模型被推导出来。在目标 3 中，我们将这些细胞系分化为多巴胺神经元，以解决 iPSC 建模的问题 可以揭示LND相关的发病时机和机制。总体而言，该项目将解决 与神经发育障碍的 iPSC 建模广泛相关的基本问题，以及 对 LND 神经行为异常的发病机制有更具体的见解。