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种基本细胞过程，在几种中可能会失调人类疾病。巨噬细胞是大多数组织中存在并采用的异质种群组织特异性功能。 mRNA转化控制在巨噬细胞和组织中的作用特定功能尚不清楚。拟议研究的广泛目标是了解如何 mRNA翻译的失调控制应力期间组织驻留的巨噬细胞功能。具体这项研究的目标是确定GCN2（通用控制不可压2）如何依赖性翻译巨噬细胞的控制会影响RBC生产和清除率中的巨噬细胞功能，并发现介导这种作用的基因。 GCN2是属于信号网络的丝氨酸/苏氨酸蛋白激酶通过翻译调节剂EIF2（真核翻译）协调细胞对营养应激的反应起始因子2）。 GCN2感觉氨基酸水平并磷酸化EIF2对氨基酸的响应不足。 P-EIF2抑制全局mRNA翻译，但矛盾地刺激了一部分的翻译关键应力反应基因（例如ATF4）（激活转录因子4）。应力反应的上调响应GCN2/EIF2信号传导的基因激活了一个转录程序，该程序可帮助细胞克服不利条件或凋亡。 GCN2功能先前已链接到重要生理和病理状况，例如记忆形成，癌症和炎症性疾病。但是，GCN2在调节组织居民巨噬细胞及其在RBC产生中的功能中的作用清除尚未表征。我们当前的模型表明GCN2控制RBC的生产通过调节巨噬细胞中的mRNA翻译在压力期间的清除。因此，我们提出以下目的是实现我们的目标：首先，我们将确定GCN2在RBC清除中的重要性巨噬细胞（AIM 1）并定义了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RBC成熟和生产（AIM 2）。最后，我们会的检查巨噬细胞通过GCN2通过巨噬细胞感测的方式（AIM 3）。为了实现这些目标，我们将使用缺乏GCN2或在巨噬细胞中携带耐磷酸EIF2的转基因小鼠在全基因组水平上研究mRNA翻译的技术。我们的小鼠模型，体内和体外实验将严格评估巨噬细胞在GCN2依赖性发展中的核心作用压力期间的缺陷。我们全基因组方法和所选目标的体外功能分析将发现在巨噬细胞中起重要作用的GCN2下游的新型翻译调节基因压力期间RBC产生的调节。