Project 3

项目3

基本信息

批准号：
10271309
负责人：
PENG JIN
金额：
$ 44.64万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-25 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10271309
关键词：
3-Dimensional Address Animal Model Behavioral Biological Assay Biological Models Brain CGG repeat Cells Cerebrum Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Cognitive Collaborations Complex Data Development Disease Drug Screening Electrophysiology (science)Evaluation Exhibits FDA approved FMR1 Failure Fragile X Syndrome GABA Receptor Gene Expression Profile Genes Genetic Genomics High-Throughput Nucleotide Sequencing Human Human Characteristics Human Development Human body Individual Inherited Intellectual functioning disability Libraries Link Mediating Messenger RNA Metabotropic Glutamate Receptors Metformin Methods Modeling Molecular Mus Names Neurons Organ Organogenesis Organoids Outcome Pathologic Pathway interactions Patients Pharmaceutical Preparations Pharmacotherapy Phase Phase III Clinical Trials Phenotype Pluripotent Stem Cells Property Publishing Regulation Reporter Ribosomal Protein S6 Kinase Role Sampling Signal Transduction Structure Testing Therapeutic Therapeutic Effect Tissues Work autism spectrum disorder base cell type disease phenotype drug development drug repurposing epigenomics functional loss gamma-Aminobutyric Acid genome editing human disease human model induced pluripotent stem cell inhibitor/antagonist interest kinase inhibitor molecular phenotype mouse model nerve stem cell new therapeutic target novel novel strategies novel therapeutic intervention pre-clinical risk variant screening small molecule stem cell model synergism therapeutic genome editing therapeutic target three dimensional cell culture three-dimensional modeling tool transcriptome sequencing treatment strategy virtual

项目摘要

Project Summary Fragile X syndrome (FXS) is the most common inherited form of intellectual disability and a leading genetic cause of autism spectrum disorders (ASD). FXS is caused by the loss of functional fragile X mental retardation protein (FMRP). Previous works have focused on the role of FMRP as a translational regulator, and many mRNA targets of FMRP have been shown to be ASD-linked genes. Despite major progress to characterize underlying disease mechanisms in animal models that has led to several clinical trials, including phase 3 clinical trials of drugs modulating metabotropic glutamate and GABA receptors, improvements of behavioral and cognitive outcomes in patients have unfortunately been largely unsuccessful. We believe that a major gap in the preclinical phase of drug development for FXS can be addressed by the development of human FXS induced pluripotent stem cell (iPSC) derived models, which will enable us to identify human specific therapeutic targets and evaluate novel therapeutic approaches. Human iPSCs are pluripotent and are able to generate many different cell types. Three-dimensional (3D) organoid culture of iPSCs has evolved from embryoid body culture, quite faithfully following human organogenesis, and provides a new platform to investigate human brain development in a dish, otherwise inaccessible to experimentation. We have developed FXS iPSC models, including 2D neural progenitor cells (NPCs)/cortical neurons and 3D cortical organoids, and identified a number of FMRP target mRNAs in the human context. Furthermore, we have observed abnormalities associated with the loss of FMRP at molecular, cellular and electrophysiological levels in FXS iPSC models. Intriguingly, our preliminary data suggest that PI3K inhibitors, but not mGluR5 antagonists, could rescue cellular phenotypes in human FXS iPSC derived model systems, potentially validating the failure of positive preclinical mouse studies with negative human trials. In this proposed study, we aim to use human specific iPSC models as translational tools to develop novel therapeutic approaches for FXS. First, we will determine the therapeutic effects of compounds targeting candidate pathways in FXS organoids (Aim 1). Second, we will develop CRISPR-based genomic and epigenomic editing therapeutic approaches to reactivate FMR1 expression in FXS organoids (Aim 2). Third, we will conduct molecular phenotype and FMR1-reactivation-based small molecule screens (Aim 3). Our proposed works will lead to the identification of novel therapeutic targets and the development of new treatment strategies for FXS.

项目摘要脆弱的X综合征（FXS）是最常见的智障形式，是领先的遗传自闭症谱系障碍的原因（ASD）。 FXS是由功能脆弱X智力低下的丧失引起的蛋白质（FMRP）。以前的作品集中在FMRP作为转化调节器和许多mRNA的作用上 FMRP的靶标已显示为ASD连接基因。尽管进展取得了重大进展动物模型中的疾病机制导致了几项临床试验，包括3期临床试验调节代谢性谷氨酸和GABA受体的药物，行为和认知的改善不幸的是，患者的结果在很大程度上没有成功。我们认为临床前的主要差距 FXS的药物开发阶段可以通过人类FXS引起的多能的发展来解决干细胞（IPSC）派生的模型，这将使我们能够识别人类特定的治疗靶标并评估新颖的治疗方法。人IPSC具有多能，并且能够生成许多不同的细胞类型。 IPSC的三维（3D）类器官培养已经从胚胎体培养中演变出来，非常忠实遵循人体器官发生，并提供了一个新的平台来调查菜肴中的人脑发育，否则无法访问实验。我们开发了FXS IPSC模型，包括2D神经祖细胞（NPC）/皮质神经元和3D皮质器官，并鉴定出许多FMRP靶标在人类背景下的mRNA。此外，我们观察到与FMRP丢失有关的异常 FXS IPSC模型中的分子，细胞和电生理水平。有趣的是，我们的初步数据建议PI3K抑制剂（而不是MGLUR5拮抗剂）可以挽救人FXS IPSC中的细胞表型衍生的模型系统，有可能验证阳性临床前小鼠研究的失败人类试验。在这项拟议的研究中，我们旨在使用人类特异性IPSC模型作为转化工具来开发 FXS的新型治疗方法。首先，我们将确定靶向化合物的治疗作用 FXS器官中的候选途径（AIM 1）。其次，我们将开发基于CRISPR的基因组和表观基因组学编辑治疗方法重新激活FXS器官中的FMR1表达（AIM 2）。第三，我们将进行基于分子表型和基于FMR1折射的小分子筛选（AIM 3）。我们提出的作品将导致鉴定新的治疗靶标和FXS的新治疗策略的发展。