Defining the role of Tox3 in congenital cerebellar hypoplasia and ataxia

定义 Tox3 在先天性小脑发育不全和共济失调中的作用

基本信息

批准号：
10799992
负责人：
Joshua John Breunig
金额：
$ 58.45万
依托单位：
CEDARS-SINAI MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-15 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10799992
关键词：
ACTL6B gene ATAC-seq Ablation Acute Anatomy Ataxia Attention deficit hyperactivity disorder Biological Models Brain Brain Diseases Brain region CD8-Positive T-Lymphocytes Calcium Signaling Cell Fate Control Cells Cerebellar Ataxia Cerebellar Diseases Cerebellum Childhood Chromium Cognition Cognitive Complement Congenital cerebellar hypoplasia Coupled DNA Binding Domain Data Development Disease Dyslexia Electroporation Elements Embryo Emotions Epigenetic Process Equilibrium Family Fourth ventricle structure Fractionation Functional disorder Gene Expression Generations Genes Genetic Goals Histones Human Image Immune system In Vitro Investigation Knock-out Knockout Mice Label Language Light Lip structure Malignant Neoplasms Malignant neoplasm of brain Maps Mediating Methods Molecular Motor Movement Mus Mutagenesis Nervous System Neuroanatomy Neurons Parkinson Disease Pathologic Pathology Pattern Phenotype Population Predisposition Pregnancy Primordium Process Prosencephalon Protein Family Proteins Regulation Repression Research Restless Legs Syndrome Role Satiation Signal Transduction Site Specific qualifier value Structure Substantia nigra structure T-Lymphocyte Tamoxifen Tissues Titrations Ventricular Work autism spectrum disorder bioinformatics pipeline cell type chromatin remodeling chronic infection cognitive function comparison control conditional knockout developmental disease dopaminergic neuron epigenetic regulation exhaustion frontal lobe gene network genome wide association study granule cell histogenesis human disease human model in vivo insight malignant breast neoplasm member motor control mouse model multimodality multiple omics mutant nerve stem cell nestin protein neural notch protein novel pars compacta precursor cell single-cell RNA sequencing spatial memory spatiotemporal transcription factor transcriptomics virtual

项目摘要

PROJECT SUMMARY / ABSTRACT The cerebellum is an exquisitely laminated structure regulating the balance, voluntary motor coordination and modulating language and cognition through connections with the forebrain. Sitting just under the forebrain, cerebellum is estimated to contain roughly 80% of the neurons in the human brain. Accordingly, cerebellar dysgenesis, pathology, or dysfunction is associated with a host of diseases or disorders, including ataxia, autism, and intellectual deficit. Most cerebellar neurons are a subtype called cerebellar granule cells (CGCs), which are generated embryonically from a structure termed the rhombic lip (RL)—a germinal zone residing at the interface between the ventricular zone (VZ) and roof plate of the fourth ventricle. The mechanisms leading to the specification of the RL germinal niche and subsequent generation of granule cells remain incompletely understood. Specifically, the epigenetic and transcriptomic changes underlying how the ventricular zone precursor cells choose the to become RL are largely unknown. We have generated a novel conditional knockout (cKO) mouse for Tox3, a member of the TOX family of transcription factors previously associated with the regulation of epigenetics in T Cells. Loss of TOX3 prior to RL specification leads to almost complete agenesis of CGCs and 100% penetrant ataxia. We hypothesize that TOX3 mediates an epigenetic switch necessary for the generation of cerebellar granule cell precursors from the rhombic lip and associated ventricular zone—the loss of which results in developmental ataxia. Using our mature bioinformatics pipelines and single-cell approaches in combination with both mouse and human models systems, we will propose to investigate the mechanisms by which Tox3 regulates cerebellar histogenesis. We propose to carry out this work in two parts. The focus of Specific Aim 1 is to interrogate the epigenetic and transcriptomic consequences of cKO of Tox3 using multimodal single-cell RNA- and ATAC-sequencing combined with single-cell Cut&Tag to determine genetic networks regulating VZ/RL precursors and CGC genesis. The main goal of Specific Aim 2 is to define the role of TOX3 on murine cerebellar lineages using inducible Cre drivers and somatic mutagenesis.

项目概要/摘要小脑是一个精致的层状结构，调节平衡、随意运动协调和通过与前脑的连接调节语言和认知，据估计，小脑包含人类大脑中大约 80% 的神经元。发育不全、病理学或功能障碍与许多疾病或病症相关，包括共济失调、大多数小脑神经元是一种称为小脑颗粒细胞（CGC）的亚型，它们是从胚胎产生的一种称为菱形唇（RL）的结构——位于心室区 (VZ) 和第四脑室顶板之间的界面。 RL 生发生态位的规范和后续颗粒细胞的产生仍然不完全具体来说，是脑室区的表观遗传和转录组变化。前体细胞选择成为 RL 的情况在很大程度上是未知的。我们已经生成了一种新的条件。 Tox3 基因敲除 (cKO) 小鼠，Tox3 是先前相关的转录因子 TOX 家族的成员 RL 规范化之前 TOX3 的缺失导致 T 细胞的表观遗传学几乎完全丧失。 CGC 发育不全和 100% 渗透性共济失调我们发现 TOX3 介导表观遗传开关。从菱形唇和相关的小脑颗粒细胞前体的产生所必需的脑室区——其缺失会导致发育性共济失调。管道和单细胞方法与小鼠和人类模型系统相结合，我们将提议研究 Tox3 调节小脑组织发生的机制。我们建议分两部分开展这项工作，具体目标 1 的重点是探究表观遗传学。使用多模式单细胞 RNA 和 ATAC 测序分析 Tox3 cKO 的转录组结果结合单细胞 Cut&Tag 确定调节 VZ/RL 前体和 CGC 的遗传网络具体目标 2 的主要目标是利用小鼠小脑谱系来定义 TOX3 的作用。诱导型 Cre 驱动程序和体细胞突变。