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

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要神经发育障碍的发病机理出于许多原因而具有挑战性。人类大脑相对无法直接评估，人类尸检和成像研究有限研究分子或细胞机制的机会，动物模型有时不会复制重要疾病特征和永生的神经元样细胞系不能用于模拟正常发育。诱导多能干细胞（IPSC）提供了一种有力的新实验工具来解决这些局限性。他们可以是由由定义突变引起的已知发育障碍的患者创建的，它们提供了可再生资源，它们可以分化为特定的细胞谱系，以提供特定物种的特定和谱系特异性的实验模型，以研究细胞和分子水平的发育机制。在当前的建议中，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。我们专注于Lesch-Nyhan病（LND）作为一种原型疾病，因为它具有许多功能，使其成为IPSC建模的异常处理的Mendelian疾病。 lnd和它的温和变体是由HPRT1基因突变引起的，导致嘌呤救助酶，低黄嘌呤 - 瓜氨酸磷酸贝糖基转移酶（HGPRT）。神经行为异常疾病与脑多巴胺神经元的发育功能障碍有关。 AIM 1专注于在各种疾病严重程度中建立良好的IPSC银行，以解决问题与IPSC模型中如何反映导致不同疾病严重程度的特定突变有关。 AIM 2重点建立一个良好的等源性IPSC银行，其中引入了特定的突变通过基因编辑方法，解决与IPSC的个体的遗传背景有关的问题模型被得出。在AIM 3中，我们将这些线区分为多巴胺神经元，以解决IPSC模型可以揭示与LND相关的发病机理的时间和机制。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理的更具体见解。