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RNA Regulatory Networks in Neuronal Cell Type Diversity and Function

神经元细胞类型多样性和功能中的 RNA 调控网络

基本信息

批准号：
10816681
负责人：
Chaolin Zhang
金额：
$ 2.95万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-12-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10816681
关键词：
Alternative Splicing Disease Fumonisin B1 Gene Expression Regulation Molecular Mus Nervous System Neurons RNA RNA-Protein Interaction Regulation cell type clinically relevant insight posttranscriptional programs

项目摘要

PROJECT SUMMARY RNA regulatory networks in neuronal cell type diversity and function The mammalian brain is probably the most complex organ in the body and its proper function requires coordination of many diverse types of excitatory and inhibitory neurons that form distinct functional circuitries. Characterization of neuronal cell types is fundamental to understand not only how the brain works, but also how specific cell types are selectively affected in multiple neuronal disorders and how this process can be reversed. A large number of neuronal cell types were recenUy defined based on their gene expression profiles in bulk and single cells, and these cell types are organized into a hierarchical cell taxonomy. Alternative splicing is a mechanism to generate multiple transcript and protein variants with distinct functions, thus providing a major driving force of the molecular diversity in mammals. The overarching goal of my research group is to understand the contribution of alternative splicing and the underlying RNA-regulatory networks in the brain and brain-related disorders. Previous work from multiple groups, including ours, unambiguously demonstrated that the brain has unique splicing-regulatory programs compared to non-neuronal tissues. Our studies also revealed the establishment of a pan-neuronal splicing program regulated by multiple RNA-binding proteins (RBPs) during neural development, and demonstrated the important role of the Rbfox protein family in regulating axonal maturation and neuronal excitability. However, how alternative splicing contributes to the distinct molecular profiles of diverse neuronal cell types in the cortex is currenUy poorly understood. We hypothesize that the transcriptome diversity generated by highly regulated alternative exons is a major component that specifies neuronal cell type identity and function. In this application, we describe our preliminary analysis of neuronal cell type-specific alternative splicing regulation in mouse cortex, which provides strong support for the following specific aims we would like to pursue: 1) Perform systematic analysis of neuronal cell type-specific alternative splicing to identify novel neuronal subclasses and regulators using deep sc-RNA-seq data. 2) Validate our computational predictions and characterize mechanisms of neuronal cell type-specific splicing regulation and function using two complementary model systems: the distinction of two major subclasses of GABAergic interneurons originating from caudal (CGE) and medial (MGE) ganglionic eminences, and a GABAergic neuronspecific microexon in Ank3/Ankyrin G. To achieve our goal, we will use a multidisciplinary approach that integrates cutting-edge statistical an machine learning methods and multiple in vitro and in vivo experimental models. This study will generate a global and mechanistic view of precise alternative splicing regulation across diverse neuronal cell types and illuminate its impact on neuronal structural and functional properties.

项目概要神经元细胞类型多样性和功能中的 RNA 调控网络哺乳动物的大脑可能是体内最复杂的器官，其正常功能需要许多不同类型的兴奋性和抑制性神经元的协调，形成不同的功能回路。神经元细胞类型的表征对于理解大脑如何工作以及如何工作至关重要特定细胞类型在多种神经元疾病中选择性地受到影响，以及如何逆转这一过程。最近根据大量的基因表达谱定义了大量的神经元细胞类型单细胞，这些细胞类型被组织成分层细胞分类法。选择性拼接是产生具有不同功能的多个转录物和蛋白质变体的机制，从而提供了主要的哺乳动物分子多样性的驱动力。我的研究小组的首要目标是了解选择性剪接的贡献以及大脑中潜在的 RNA 调控网络和大脑相关失调。包括我们在内的多个小组之前的工作明确表明，大脑已经与非神经元组织相比，独特的剪接调节程序。我们的研究还揭示了建立由多种RNA结合蛋白（RBP）调控的泛神经元剪接程序神经发育，并证明了 Rbfox 蛋白家族在调节轴突中的重要作用成熟度和神经元兴奋性。然而，选择性剪接如何有助于形成独特的分子目前人们对皮质中不同神经元细胞类型的概况知之甚少。我们假设由高度监管的替代外显子产生的转录组多样性是指定的主要组成部分神经元细胞类型的识别和功能。在此应用中，我们描述了对神经元细胞的初步分析小鼠皮层中类型特异性选择性剪接调控，为以下方面提供了强有力的支持我们想要追求的具体目标：1）对神经元细胞类型特异性替代方案进行系统分析使用深度 sc-RNA-seq 数据进行剪接以识别新的神经元亚类和调节因子。 2）验证我们的计算预测和表征神经元细胞类型特异性剪接调节的机制和使用两个互补模型系统的函数：GABAergic 的两个主要子类的区别源自尾侧 (CGE) 和内侧 (MGE) 神经节隆起的中间神经元，以及 GABA 能神经元特异性 Ank3/Ankyrin G 中的微外显子。为了实现我们的目标，我们将使用多学科方法整合了尖端的统计和机器学习方法以及多种体外和体内实验模型。这项研究将对跨领域的精确选择性剪接调控产生全局和机械的观点。不同的神经元细胞类型，并阐明其对神经元结构和功能特性的影响。