Translational regulation of tissue resident macrophages by GCN2

GCN2 对组织驻留巨噬细胞的翻译调节

基本信息

批准号：
10611500
负责人：
Soroush Tahmasebi
金额：
$ 47.97万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
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：
2022-05-01 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10611500
关键词：
Address Adhesives Adopted Affect Amino Acids Animal Model Apoptosis Binding Proteins Biochemical Biomedical Engineering Bone Marrow Candidate Disease Gene Cardiac Cell physiology Cells ChIP-seq Chicago Collaborations Consultations Data Defect Development Disease EIF-2alpha Erythroblasts Erythrocytes Erythroid Erythrophagocytosis Erythropoiesis Eukaryotic Initiation Factors Fetal Liver Gene Expression Genes Genetic Transcription Goals Heme High Fat Diet Homeostasis In Vitro Inflammatory Knock-in Mouse Knock-out Knockout Mice Knowledge Letters Life Cycle Stages Link Macrophage Malignant Neoplasms Mammalian Cell Mechanical Stress Mediating Memory Messenger RNA Modeling Molecular Mus Nutrient Pathologic Pathway interactions Phagocytes Phagocytosis Phosphorylation Phosphotransferases Physiological Piezo 1 ion channel Play Population Heterogeneity Process Production Prokaryotic Initiation Factor-2 Protein Biosynthesis Protein-Serine-Threonine Kinases Regulation Resistance Resolution Ribosomes Role Scientist Seminal Signal Pathway Signal Transduction Stress Techniques Technology Testing Time Tissue Engineering Tissues Transgenic Mice Translation Initiation Translational Regulation Translations Universities Up-Regulation activating transcription factor 4 biological adaptation to stress cell type experience experimental study gene network genome-wide human disease in vivo inhibitor innovation invention mRNA Translation mechanical force mouse model novel pathogen predictive modeling programs response tissue repair transcription factor translatome

项目摘要

SUMMARY mRNA translation, or protein synthesis, is a fundamental cellular process that can be dysregulated in several human diseases. Macrophages are heterogeneous populations that are present in most tissues and adopt tissue specific functions. The role of mRNA translational control in macrophages and in regards to their tissue specific functions is not well understood. The broad goal of the proposed studies is to understand how dysregulation of mRNA translation controls tissue-resident macrophage function during stress. The specific goals of this study are to identify how GCN2 (general control nonderepressible 2)-dependent translational control in macrophages affects macrophage function in RBC production and clearance and to uncover the genes mediating this effect. The GCN2 is a serine/threonine-protein kinase that belongs to a signaling network that coordinates cellular response to nutrient stress through translational regulator eIF2 (Eukaryotic translation initiation factor 2). GCN2 senses amino acid levels and phosphorylates eIF2 in response to amino acid deficiency. p-eIF2 inhibits global mRNA translation but paradoxically stimulates the translation of a subset of key stress-response genes such as ATF4 (Activating Transcription Factor 4). Upregulation of stress-response genes in response to GCN2/eIF2 signaling activates a transcription program that helps the cells to overcome unfavorable conditions or undergo apoptosis. GCN2 function has been previously linked to important physiological and pathological conditions such as memory formation, cancer and inflammatory diseases. However, the role of GCN2 in regulating tissue-resident macrophages and their functions in RBC production and clearance has not been characterized. Our current model suggests that GCN2 controls RBC production and clearance during stress through regulation of mRNA translation in macrophages. Therefore, we propose the following aims to achieve our goals: First we will determine the importance of GCN2 in RBC clearance by macrophages (Aim 1) and define molecular mechanisms through which GCN2 impact this process. Next, we will elucidate how GCN2 controls RBC maturation and production by macrophages (Aim 2). Finally, we will examine how mechanical force sense by macrophage through GCN2 (Aim 3). To achieve these goals we will use transgenic mice lacking GCN2 or carrying phospho-resistant eIF2 in macrophages and state-of-art technology to study mRNA translation at genome-wide level. Our mouse models and in vivo and in vitro experiments will rigorously assess the central role of macrophages in development of GCN2-dependent defects during stress. Our genome-wide approach and in vitro functional analysis of selected targets will discover novel translationally regulated genes downstream of GCN2 that play important roles in macrophage regulation of RBC production during stress.

概括 mRNA 翻译或蛋白质合成是一个基本的细胞过程，可能在多种情况下失调。人类疾病。巨噬细胞是存在于大多数组织中的异质群体，并采用组织特定功能。 mRNA 翻译控制在巨噬细胞及其组织中的作用具体功能不太了解。拟议研究的总体目标是了解如何 mRNA 翻译失调控制应激期间组织驻留的巨噬细胞功能。具体的本研究的目标是确定 GCN2（一般控制非去抑制 2）依赖性翻译如何巨噬细胞的控制会影响巨噬细胞在红细胞生成和清除中的功能，并揭示巨噬细胞的功能介导这种效应的基因。 GCN2 是一种丝氨酸/苏氨酸蛋白激酶，属于信号网络通过翻译调节因子 eIF2（真核翻译）协调细胞对营养应激的反应引发因素2)。 GCN2 感知氨基酸水平并响应氨基酸磷酸化 eIF2 不足。 p-eIF2 抑制整体 mRNA 翻译，但矛盾的是刺激了一部分 mRNA 的翻译关键的应激反应基因，例如 ATF4（激活转录因子 4）。应激反应的上调响应 GCN2/eIF2 信号的基因会激活转录程序，帮助细胞克服不利条件或发生细胞凋亡。 GCN2 功能之前已被认为与重要的生理和病理状况，例如记忆形成、癌症和炎症性疾病。然而，GCN2 在调节组织驻留巨噬细胞中的作用及其在红细胞生成中的功能并且间隙尚未被表征。我们当前的模型表明 GCN2 控制红细胞的产生以及通过巨噬细胞中 mRNA 翻译的调节在应激期间进行清除。因此，我们建议为了实现我们的目标，我们将采取以下措施：首先，我们将通过以下方式确定 GCN2 在 RBC 清除中的重要性：巨噬细胞（目标 1）并定义 GCN2 影响该过程的分子机制。接下来，我们将阐明 GCN2 如何控制巨噬细胞的红细胞成熟和生成（目标 2）。最后，我们将检查巨噬细胞如何通过 GCN2 感知机械力（目标 3）。为了实现这些目标，我们将使用缺乏 GCN2 或巨噬细胞中携带磷酸抗性 eIF2 的转基因小鼠和最先进的技术在全基因组水平上研究 mRNA 翻译的技术。我们的小鼠模型以及体内和体外实验将严格评估巨噬细胞在 GCN2 依赖性发育中的核心作用压力期间的缺陷。我们对选定目标的全基因组方法和体外功能分析将发现 GCN2 下游的新型翻译调控基因，这些基因在巨噬细胞中发挥重要作用应激期间红细胞生成的调节。