Olfactory neuroepithelial cell derived induced neurons for study of schizophrenia

嗅觉神经上皮细胞衍生的诱导神经元用于精神分裂症研究

• 批准号: 10747189
• 负责人: Karin Borgmann-Winter
• 金额: $ 47.58万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10747189
• 关键词: ASCL1 gene; ATAC-seq; Age; Animals; Bar Codes; Brain; Bypass; Cell Differentiation process; Cell Nucleus; Cells; Cellular Assay; Characteristics; Chromatin; Complex; Development; Disease; Electrophysiology (science); Epigenetic Process; Evolution; Fibroblasts; Functional disorder; Generations; Genes; Genetic; Genetic Transcription; Genetic Variation; Goals; Growth; In Vitro; Individual; Induced pluripotent stem cell derived neurons; Mental disorders; Methodology; Methods; MicroRNAs; Modeling; Modification; Molecular; Molecular Conformation; Molecular Genetics; Mood Disorders; Neurobiology; Neuroepithelial; Neuroepithelial Cells; Neuronal Differentiation; Neurons; Organoids; Pathway interactions; Patients; Phenotype; Pluripotent Stem Cells; Predisposition; Process; Protocols documentation; Psychotic Disorders; RNA analysis; Reporter; Research; Schizophrenia; Shapes; Skin; Somatic Cell; Source; System; Testing; Transcript; cell type; developmental disease; disease phenotype; epigenetic variation; falls; genetic variant; human subject; in vitro Model; induced pluripotent stem cell; multiple omics; neural; neuropsychiatry; novel; programs; risk variant; sex; stem; stem cell model; stem cells; theories; transcription factor; transcriptome; transcriptome sequencing

PROJECT SUMMARY The pathophysiology of common psychiatric illnesses such as schizophrenia is highly complex and heterogeneous, in which multitudes of common and rare genetic variants and their epigenetic modifications precipitate disease phenotypes. Over the last decade, in vitro modeling of neuropsychiatric illnesses has been led by induced pluripotent stem cell (iPSC) based paradigms. These approaches have a special strength in offering developmental readouts of genetic variations in neurons and organoids. However, the ability to recapitulate complex phenotypes of the illness is hampered because epigenetic characteristics of donors are erased by the stem generation process. The goal of this project is to develop and test directly-converted induced neurons (called DCiNs henceforth) for their ability to manifest neurobiological characteristics of schizophrenia. DCiNs are generated by bypassing the stem cell stage and are thus more likely to harbor epigenetic characteristics of donors. The extent to which DCiNs cells can recapitulate neurobiological phenotypes of donors, however, will vary depending on the epigenetic landscape of the source cells and the methodology for conversion of source cells into neurons. Presently, skin fibroblasts (SFs) are commonly used source cells for iN paradigms, although SFs, as non-neural cells, may not manifest epigenetic characteristics of donors with neuropsychiatric illnesses. We propose using olfactory neuroepithelial (OE) cells, the only neural cells readily obtainable from human subjects, as source cells for DCiNs paradigms. Importantly, the chromatin accessibility of OEs is vastly different from SFs, particularly around neural genes. Among various iN paradigms, we will test a transcriptional modulation-based approach, specifically using a) two transcription factors, ASCL1 and NGN2 (N2A) and b) the transcriptional effector, microRNA (mir) 9/9*-124. We will first characterize the source cells (OE vs. SF), in combination with transcriptional modulators (N2A vs. mir 9/9*- 124), for the ability to differentiate into neurons and recapitulate the epigenetic landscape of donors’ neural cells (Aim 1). As a test of the utility of this paradigm for study of common neuropsychiatric illnesses, we will apply this model to schizophrenia by examining DCiNs-OEs from patients with schizophrenia and controls to delineate the extent to which they recapitulate disease-associated changes (Aim 2). The results of this project will define the extent to which DCiNs -OEs recapitulate neurobiological phenotypes of donors’ and pave the path for establishing the iN paradigms for other neuropsychiatric illnesses.

项目摘要 精神分裂症等常见精神病的病理生理非常复杂，并且 异质，其中众多常见和稀有遗传变异及其表观遗传修饰 沉淀疾病表型。在过去的十年中，神经精神疾病的体外建模一直是 由诱导多能干细胞（IPSC）范式领导。这些方法具有特殊的优势 提供神经元和器官遗传变异的读数。但是，能够 概括疾病的复杂表型受到了阻碍，因为供体的表观遗传特征是 被STEM生成过程删除。该项目的目的是开发和测试直接转换 诱导神经元（称为DCINS），其能力表现出神经生物学特征的能力 精神分裂症。 DCIN是通过绕过干细胞阶段而产生的，因此更有可能藏有 捐助者的表观遗传特征。 DCINS细胞可以概括神经生物学的程度 但是，供体的表型将根据源细胞和源细胞的表观遗传景观而有所不同 将源细胞转化为神经元的方法。目前，通常使用皮肤成纤维细胞（SF） 范式中的源细胞，尽管SFS（作为非神经细胞）可能没有表现出表观遗传特征 神经精神疾病的供体。我们建议使用嗅觉神经上皮（OE）细胞，这是唯一的神经元 细胞很容易从人类受试者中获得，作为DCINS范式的源细胞。重要的是，染色质 OE的可访问性与SFS大不相同，尤其是在神经基因周围。在各种in 范式，我们将测试一种基于转录调制的方法，特别是使用a）两次转录 因素，ASCL1和NGN2（N2A）和B）转录效应子MicroRNA（MIR）9/9*-124。我们将首先 表征源细胞（OE与SF），结合转录调节剂（N2A与miR 9/9* - 124），为了区分神经元并概括供体神经元的表观遗传景观 细胞（目标1）。作为该范式研究常见神经精神疾病的实用性的测试，我们将 通过检查精神分裂症患者的DCINS-OE，将此模型应用于精神分裂症 描述它们概括与疾病相关的变化的程度（AIM 2）。这个项目的结果 将定义DCINS -OE概括供体的神经生物学表型的程度，并铺平路径 用于建立其他神经精神疾病的范式。