Olfactory neuroepithelial cell derived induced neurons for study of schizophrenia

项目总结 精神分裂症等常见精神疾病的病理生理学非常复杂， 异质性，其中大量常见和罕见的遗传变异及其表观遗传修饰 沉淀疾病表型。在过去的十年里，神经精神疾病的体外建模一直是 以诱导多能干细胞(IPSC)为基础的范例。这些方法在以下方面具有特殊的优势 提供神经元和器官中遗传变异的发育读数。然而，能够 概括疾病的复杂表型受阻，因为捐赠者的表观遗传特征 通过茎的生成过程擦除。该项目的目标是开发和测试直接转换 诱导神经元(以下称为DCins)具有表现出神经生物学特性的能力 精神分裂症。DCiN是通过绕过干细胞阶段产生的，因此更有可能 捐献者的表观遗传学特征。DCins细胞可以在多大程度上重现神经生物学 然而，捐赠者的表型将根据源细胞的表观遗传格局和 将源细胞转化为神经元的方法学。目前，皮肤成纤维细胞(SFS)被广泛使用 尽管作为非神经细胞的SFS可能不表现出表观遗传学特征，但在范例中的来源细胞 患有神经精神疾病的捐赠者。我们建议使用嗅神经上皮(OE)细胞，这是唯一的神经 从人类受试者身上容易获得的细胞，作为DCins范例的来源细胞。重要的是，染色质 OES的可及性与SFS有很大的不同，特别是在神经基因方面。在各种In中 范例，我们将测试基于转录调控的方法，特别是使用a)两个转录 因子，ASCL1和Ngn2(N2a)和b)转录效应因子，microRNA(Mir)9/9*-124。我们将首先 表征源细胞(OE与SF)，结合转录调节因子(N2A与MIR 9/9*- 124)，用于分化为神经元并概括捐赠者神经的表观遗传图景 细胞(目标1)。作为对这一范例对常见神经精神疾病研究效用的测试，我们将 通过检测精神分裂症患者和对照组的DCins-Oes，将该模型应用于精神分裂症 勾勒出它们概括与疾病相关的变化的程度(目标2)。这个项目的成果 将定义DCins-Oes在多大程度上概括了捐赠者的神经生物学表型，并为 为其他神经精神疾病建立In范例。

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.