Human Induced Pluripotent Stem Cells As Models for Inherited Developmental Disorders

人类诱导多能干细胞作为遗传性发育障碍的模型

• 批准号: 9512060
• 负责人: HYDER A JINNAH
• 金额: $ 54.6万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9512060
• 关键词: Address; Affect; Animal Model; Attenuated; Autopsy; Biochemical; Biochemical Pathway; Biological Process; Biological Response Modifier Therapy; Biology; Brain; CRISPR/Cas technology; Cell Line; Cell Lineage; Cell model; Cells; Clinical; Defect; Development; Developmental Process; Disease; Enzymes; Epigenetic Process; Etiology; Evaluation; Experimental Models; Feasibility Studies; Fibroblasts; Future; Gene Expression; Generations; Genes; Genetic; HPRT1 gene; Human; Hypoxanthine Phosphoribosyltransferase; In Vitro; Inherited; Intervention; Knock-out; Lesch-Nyhan Syndrome; Link; Measurement; Mendelian disorder; Methods; Modeling; Molecular; Morphology; Mutation; Neurobiology; Neurodevelopmental Disorder; Neuronal Differentiation; Neuronal Dysfunction; Neurons; Outcome; Pathogenesis; Pathway interactions; Patients; Pharmacology; Pharmacology Study; Phenotype; Pilot Projects; Process; Proteome; Proteomics; Purines; Resources; Role; Severities; Severity of illness; Technology; Testing; Therapeutic Intervention; Variant; base; clinical phenotype; clinically relevant; developmental disease; dopaminergic neuron; enzyme deficiency; gene product; gene therapy; imaging study; immortalized cell; individual patient; induced pluripotent stem cell; metabolome; metabolomics; neurite growth; neurobehavioral; neurodevelopment; novel; protein expression; prototype; purine metabolism; relating to nervous system; restoration; tool; transcriptome; transcriptomics

PROJECT SUMMARY The mechanisms responsible for neural dysfunction in many neurodevelopmental disorders are 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 mechanism, animal models sometimes do not replicate key features, and immortalized neuron-like cell models have uncertain value for studying normal developmental processes. Induced pluripotent stem cells (iPSCs) provide a powerful new experimental tool to address these limitations, because they can be created from patients with defined mutations, and they can be differentiated in vitro into specific cell lineages to provide species-specific and lineage-specific cell models. In the current proposal we describe the development and characterization of a resource of iPSCs to serve as experimental cell-based models for Lesch-Nyhan disease (LND) and its attenuated variants. These disorders are caused by mutations in the HPRT1 gene, resulting in deficiency of the purine salvage enzyme, hypoxanthine-guanine phosphoribosyltransferase (HGprt). LND provides an exceptionally tractable Mendelian disorder for iPSC modeling. Because this technology has not yet been methodically exploited for this disorder, the main focus of the current proposal is the development and initial characterization of a resource of iPSCs for discovery of novel biological processes and therapeutic interventions. Specifically, we will establish a well- characterized and clinically annotated bank of iPSCs from patients with a spectrum of disease severity as well as isogenic lines in which specific mutations have been introduced via gene editing methods (Aim 1); characterize morphological defects associated with differentiation of these iPSCs into neurons (Aim 2); determine the molecular and biochemical pathways affected in these cells using comprehensive transcriptomics, proteomics and metabolomics (Aim 3); and determine how early in the developmental process HGprt must be restored to correct neuronal defects (Aim 4). This project will result in a novel and critically important resource for future studies of the pathogenesis of LND and future studies of pharmacological or genetic therapeutic interventions.

项目概要 许多神经发育障碍中神经功能障碍的机制对于科学家来说具有挑战性。 地址有很多原因。人类大脑相对难以直接评估、人体尸检 和成像研究提供的研究机制的机会有限，动物模型有时不提供 复制关键特征，永生化神经元样细胞模型对于研究正常细胞具有不确定的价值 发展过程。诱导多能干细胞（iPSC）提供了一种强大的新实验工具 解决了这些限制，因为它们可以从具有明确突变的患者身上产生，并且可以 在体外分化为特定的细胞谱系，以提供物种特异性和谱系特异性的细胞模型。在 在当前的提案中，我们描述了 iPSC 资源的开发和表征，以充当 Lesch-Nyhan 病 (LND) 及其减毒变种的实验性细胞模型。这些疾病 由 HPRT1 基因突变引起，导致嘌呤补救酶缺乏， 次黄嘌呤鸟嘌呤磷酸核糖转移酶（HGprt）。 LND 提供了一种非常容易处理的孟德尔 iPSC 建模障碍。由于这项技术尚未被系统地用于治疗这种疾病， 当前提案的主要重点是 iPSC 资源的开发和初步表征 发现新的生物过程和治疗干预措施。具体来说，我们将建立一个完善的 来自具有一系列疾病严重程度的患者的特征化和临床注释的 iPSC 库 作为通过基因编辑方法引入特定突变的同基因系（目标 1）； 表征与这些 iPSC 分化为神经元相关的形态缺陷（目标 2）； 使用综合方法确定这些细胞中受影响的分子和生化途径 转录组学、蛋白质组学和代谢组学（目标 3）；并确定在发育过程的早期阶段 必须恢复 HGprt 以纠正神经元缺陷（目标 4）。该项目将产生一种新颖且具有批判性的 未来 LND 发病机制研究和药理学或未来研究的重要资源 基因治疗干预。

项目成果

Lesch-Nyhan disease causes impaired energy metabolism and reduced developmental potential in midbrain dopaminergic cells.

• DOI: 10.1016/j.stemcr.2021.06.003
• 发表时间: 2021-07-13
• 期刊: Stem cell reports
• 影响因子: 5.9
• 作者: Bell S;McCarty V;Peng H;Jefri M;Hettige N;Antonyan L;Crapper L;O'Leary LA;Zhang X;Zhang Y;Wu H;Sutcliffe D;Kolobova I;Rosenberger TA;Moquin L;Gratton A;Popic J;Gantois I;Stumpf PS;Schuppert AA;Mechawar N;Sonenberg N;Tremblay ML;Jinnah HA;Ernst C
• 通讯作者: Ernst C

Macrocytic anemia in Lesch-Nyhan disease and its variants.

• DOI: 10.1038/s41436-018-0053-1
• 发表时间: 2019-03
• 期刊: Genetics in medicine : official journal of the American College of Medical Genetics
• 作者: Cakmakli HF;Torres RJ;Menendez A;Yalcin-Cakmakli G;Porter CC;Puig JG;Jinnah HA
• 通讯作者: Jinnah HA