Discover the signaling basis for OPC homeostasis

发现 OPC 稳态的信号传导基础

基本信息

批准号：
10525872
负责人：
Hui Zong
金额：
$ 45.31万
依托单位：
UNIVERSITY OF VIRGINIA
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：
2022-08-09 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10525872
关键词：
Address Apoptosis Biological Biological Assay Biology Brain Candidate Disease Gene Cell Count Cell Culture Techniques Cell Cycle Cell Density Cell Lineage Cell Proliferation Cell Survival Cell division Cells Coculture Techniques Data Data Set Degenerative Disorder Dissociation Ensure Ephrins Equilibrium Event Glioma Gliomagenesis Growth Factor Health Homeostasis Label Ligands Light Malignant Glioma Malignant Neoplasms Mediating Modeling Molecular Mosaicism NF1 gene NF1 mutation Nature Network-based Neuroglia Noise Pathway interactions Patients Pattern Phosphopeptides Platelet-Derived Growth Factor alpha Receptor Proteins Proteomics Reagent Reporting Role Siblings Side Signal Transduction Stable Isotope Labeling System TP53 gene Testing Therapeutic Wild Type Mouse Work Yang Yin Yin-Yang base candidate validation cell regeneration cell type experimental study insight intravital imaging mouse genetics mouse model mutant myelin degeneration novel strategies oligodendrocyte precursor phosphoproteomics precursor cell prevent statistical learning success tumor unsupervised learning

项目摘要

In this proposal, we will address a fundamentally important problem in glia biology: how do oligodendrocyte precursor cells (OPCs) robustly maintain their numbers? Previous intravital imaging experiments demonstrated that OPCs exert mutual inhibition when they contact each other, but promptly enter cell cycle when neighboring OPCs either differentiate or die, and halt proliferation again when homeostasis is achieved. However, the molecular basis behind OPC homeostasis remains largely unknown. Recently, using a mouse genetic system called Mosaic Analysis of Double Markers (MADM) to model glioma, our lab discovered that OPC is a cell of origin for glioma, and revealed that, instead of passively over-expanding, mutant OPCs actually outcompete WT OPCs and eventually take over the entire brain. Most importantly, when we genetically blocked cell competition of mutant OPCs, glioma can be completely prevented. Putting the observations of OPC homeostasis in health and OPC competition in gliomagenesis together, we realized that these are the two sides of the same coin, and that there is a Yin/Yang mechanism for OPC proliferation that counterbalances each other in health but gets deregulated in cancer. While the Yang network of RTK signaling is well studied, the Yin network is much less understood, let alone their counter-interactions. Here, we hypothesize that phospho-proteomic/proteomic profiling and subsequent candidate validation using highly sensitive OPC competition platforms should enable us to provide mechanistic insights into this Yin/Yang network. To test this hypothesis, we have assembled a team of experts on OPC purification and culture, proteomic profiling with limited materials, and using advanced statistical and unsupervised learning approaches to predict signaling network based on phospho-proteomic/proteomic profiles. As a team, we have successfully performed a pilot experiment that led to a handful of candidate genes. In Aim 1 of this proposal, we will validate the role of these candidate genes in OPC competition. In Aim 2 of this proposal, we will perform further in-depth profiling experiments to gain a comprehensive insight into the signaling network that controls OPC homeostasis and competition. The findings from our project should motivate further functional studies to clearly delineate the entire pathway, deepen our understanding of OPC homeostasis, and shed light on paradigm-shifting therapeutic strategies for glioma based on the concept of OPC competition.

在此提案中，我们将解决神经胶质生物学的根本重要问题：少突胶质细胞如何前体细胞（OPC）可靠地保持其数量？以前的插入成像实验证明了当他们相互接触时，OPC会施加相互抑制，但在相邻时立即进入细胞周期 OPC可以区分或死亡，并在实现稳态时再次停止增殖。但是， OPC稳态背后的分子基础在很大程度上仍然未知。最近，使用小鼠遗传系统我们的实验室发现了对神经胶质瘤的模型，称为双标记（MADM），发现OPC是一个细胞神经胶质瘤的起源，并揭示了突变的OPC实际上超过了，而不是被动过度膨胀 WT OPC，最终接管了整个大脑。最重要的是，当我们遗传阻断细胞时突变OPC的竞争可以完全预防神经胶质瘤。提出OPC的观察神经胶质作用的健康和OPC竞争中的体内平衡，我们意识到这是两个同一枚硬币的一面，并且有一个阳光/杨机构用于OPC增殖，以平衡彼此在健康方面，但在癌症中受到管制。虽然对RTK信号的Yang网络进行了充分的研究，但阴网络的理解少得多，更不用说它们的反互动了。在这里，我们假设使用高度敏感的OPC，磷酸化 - 蛋白质组/蛋白质组学分析和随后的候选验证竞争平台应使我们能够为这个阴/阳网络提供机械见解。测试这个假设，我们召集了一个专家团队OPC纯化和文化，蛋白质组学分析与材料有限，并使用先进的统计和无监督的学习方法来预测信号基于磷酸蛋白质组/蛋白质组谱的网络。作为一个团队，我们成功地执行了飞行员导致少数候选基因的实验。在本提案的目标1中，我们将验证这些作用 OPC竞争中的候选基因。在该提案的目标2中，我们将进行进一步的深入分析实验以获得控制OPC稳态和的信号网络的全面见解竞赛。我们项目的发现应激发进一步的功能研究，以清楚地描述整个道路，加深我们对OPC稳态的理解，并在范式转移时阐明基于OPC竞争概念的神经胶质瘤治疗策略。