Novel Functions of OLIG2 in Regulating Human Interneuron Production in Health and Disease

OLIG2 在健康和疾病中调节人类中间神经元产生的新功能

基本信息

批准号：
9906920
负责人：
Peng Jiang
金额：
$ 31.54万
依托单位：
RUTGERS, THE STATE UNIV OF N.J.
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-04-01 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9906920
关键词：
Animal Model Brain CRISPR/Cas technology Cell Transplantation Cells Chromosome 21 Cognitive deficits Collaborations Comb animal structure Development Disease Down Syndrome Electrophysiology (science)Embryo Embryonic Development Equilibrium Family Ganglia Gene Dosage Generations Genes Health Human Human Chromosomes Intellectual functioning disability Interneurons Knock-out Light Link Maps Molecular Biology Mus Mutation Neurons Patients Pattern Population Production Property Prosencephalon Rodent Role Structure Synaptic plasticity Syndrome System Technology Telencephalon Testing Therapeutic Transplantation Trisomy brain tissue causal variant genome editing in vivo induced pluripotent stem cell insight mouse model nerve stem cell nervous system disorder neural circuit novel overexpression progenitor relating to nervous system transcription factor

项目摘要

Disruption of excitatory/inhibitory balance (E/I imbalance) is one of the underlying causes of cognitive deficit of Down syndrome (DS), a neurodevelopmental disease characterized by triplication of human chromosome 21 (HSA21). This E/I imbalance in DS is largely resulted from overproduction of GABAergic interneurons. To study the mechanisms of intellectual disability of DS, we must better understand how interneuron production from neural progenitor cells (NPCs) is regulated during development. OLIG genes, including OLIG1 and OLIG2, are mapped to HSA21 and triplicated in DS. Studies in mouse models demonstrate that during embryonic development, both Olig1 and 2 are abundantly expressed in the ganglionic eminence, a brain structure located in the ventral embryonic telencephalon and from where most cortical interneurons are born. Moreover, Olig genes critically regulate interneuron production. Notably, expression of OLIG genes is starkly different in the human versus rodent developing ganglionic eminence. We found that differentiation of human induced pluripotent stem cell (hiPSCs) to ventral forebrain NPCs recapitulated the previous findings in human brain tissue that OLIG2 was expressed in a subpopulation of human NPCs in the ganglionic eminence. In contrast, OLIG1 was expressed in very few of these NPCs and had a complimentary expression pattern with OLIG2. Up to now, the functions of human OLIG genes in the development of human GABAergic neuron is largely unknown. Using DS patient-derived hiPSCs, we further identified that OLIG2 was overexpressed in the DS hiPSC-derived ventral forebrain NPCs. Therefore, we hypothesize that abnormal expression of OLIG genes, particularly OLIG2, in human DS ventral forebrain NPCs determines the overproduction of GABAergic neurons from these progenitors, which leads to E/I imbalance and significantly contributes to intellectual disability of DS. To test the hypothesis, I proposed three specific aims. Aim 1: to determine the role of OLIG2 in regulating human GABAergic neuron production from normal hiPSCs. We will employ OLIG2 knockout hiPSCs generated by using CRISPR/Cas9 technology to study the role of OLIG2 in human interneuron development. Aim 2: to determine whether OLIG2 is a causal gene of the overproduction of GABAergic neurons in DS. By using control and DS hiPSCs, as well as the DS hiPSCs with normalized OLIG2 gene dosage, we will determine whether overexpression of OLIG2 causes the overproduction of GABAergic neurons in DS. Aim 3: by using a novel humanized neuronal chimeric mouse model, we will further examine interneuron specification of the normal and DS hiPSC-derived ventral forebrain NPCs and their integration and contribution to E/I imbalance in vivo within intact neural circuits. Findings from this proposed study will provide novel insights into the function of OLIG2 in regulating human GABAergic neuron production and shed new light on developing potential therapeutic applications for DS by regulating the expression of OLIG2 gene.

兴奋/抑制平衡的破坏（E/I失衡）是认知不足的根本原因之一唐氏综合症（DS），一种神经发育疾病，其特征是人类染色体21 （HSA21）。 DS中的E/I不平衡主要是由于GABA能中间神经元的过量产生。到研究DS智力残疾的机制，我们必须更好地了解中间神经元如何产生在发育过程中调节神经祖细胞（NPC）。 Olig基因，包括Olig1和Olig2，被映射到HSA21，并在DS中进行了一式定链。小鼠模型的研究表明，在胚胎中开发，olig1和2均在神经节杰出中大量表达，这是一种位于大脑结构在腹胚胎端脑以及大多数皮质中间神经元出生的地方。而且，olig 基因严重调节中间神经元的产生。值得注意的是，olig基因的表达在人与啮齿动物发展神经节的杰出。我们发现人类诱发的分化多能干细胞（HIPSC）到腹前脑NPC概括了人脑的先前发现 Olig2的组织在神经节杰出中人类NPC的亚群中表达。相比之下， OLIG1在这些NPC中很少表达，并且具有Olig2的免费表达模式。向上到现在，人类olig基因在人GABA能神经元发展中的功能在很大程度上是未知。使用DS患者来源的HIPSC，我们进一步确定了DS中的Olig2过表达 HIPSC衍生的腹前脑NPC。因此，我们假设olig基因的异常表达，特别是Olig2，在人类DS腹侧前脑NPC中决定了GABA能神经元的过量生产从这些祖细胞导致E/I不平衡的祖细胞中，显着导致DS的智力残疾。为了检验假设，我提出了三个特定目标。目标1：确定olig2在调节中的作用正常HIPSC的人GABA能神经元产生。我们将使用OLIG2敲除HIPSC生成通过使用CRISPR/CAS9技术来研究Olig2在人间神经元开发中的作用。目标2：到确定olig2是否是DS中GABA能神经元过度生产过多的因果基因。通过使用对照和DS HIPSC以及具有归一化olig2基因剂量的DS HIPSC，我们将确定 Olig2的过表达是否会导致DS中GABA能神经元的过量产生。目标3：使用新型人性化神经嵌合小鼠模型，我们将进一步研究正常和DS HIPSC衍生的腹前脑NPC及其对E/I不平衡的贡献完整的神经回路内的体内。这项拟议的研究的发现将为功能提供新的见解 Olig2在调节人Gabaergic神经元产生中，并为发展潜力提供了新的启示通过调节Olig2基因的表达来用于DS的治疗应用。