Project 3

项目3

基本信息

批准号：
10443849
负责人：
PENG JIN
金额：
$ 44.73万
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/10443849
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 antagonist 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 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.

项目概要脆性 X 综合征 (FXS) 是智力障碍最常见的遗传形式，也是一种主要的遗传性疾病自闭症谱系障碍（ASD）的原因。 FXS是由于功能性脆性X智力障碍丧失而引起的蛋白质（FMRP）。之前的工作主要集中在 FMRP 作为翻译调节因子的作用，以及许多 mRNA FMRP 的目标已被证明是 ASD 相关基因。尽管在描述潜在特征方面取得了重大进展动物模型中的疾病机制已导致多项临床试验，包括 3 期临床试验调节代谢型谷氨酸和 GABA 受体的药物，改善行为和认知不幸的是，患者的治疗结果基本上不成功。我们认为临床前研究存在重大差距 FXS 药物开发阶段可以通过开发人类 FXS 诱导多能来解决干细胞（iPSC）衍生模型，这将使我们能够识别人类特定的治疗靶点并评估新的治疗方法。人类 iPSC 具有多能性，能够产生许多不同的细胞类型。 iPSC 的三维 (3D) 类器官培养是从类胚体培养发展而来，非常忠实地遵循人类器官发生，并提供一个新的平台来研究培养皿中人类大脑的发育，否则无法进行实验。我们开发了 FXS iPSC 模型，包括 2D 神经网络祖细胞 (NPC)/皮质神经元和 3D 皮质类器官，并确定了许多 FMRP 靶点人类环境中的 mRNA。此外，我们还观察到与 FMRP 丢失相关的异常情况 FXS iPSC 模型中的分子、细胞和电生理水平。有趣的是，我们的初步数据表明 PI3K 抑制剂（而非 mGluR5 拮抗剂）可以挽救人类 FXS iPSC 中的细胞表型衍生的模型系统，可能验证阳性临床前小鼠研究的失败与阴性人体试验。在这项拟议的研究中，我们的目标是使用人类特定的 iPSC 模型作为转化工具来开发 FXS 的新治疗方法。首先，我们将确定靶向化合物的治疗效果 FXS 类器官中的候选途径（目标 1）。其次，我们将开发基于CRISPR的基因组和表观基因组编辑治疗方法以重新激活 FXS 类器官中的 FMR1 表达（目标 2）。三、我们将进行分子表型和基于 FMR1 重新激活的小分子筛选（目标 3）。我们提出的作品将导致 FXS 新治疗靶点的确定和新治疗策略的开发。