MAP2K1 AND MAP2K2 IN ACUTE LUNG INJURY AND RESOLUTION

MAP2K1 和 MAP2K2 在急性肺损伤中的作用及缓解

基本信息

批准号：
10741574
负责人：
Anne M. Manicone
金额：
$ 73.65万
依托单位：
UNIVERSITY OF WASHINGTON
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：
2023-08-24 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10741574
关键词：
Acceleration Acute Lung Injury Acute Respiratory Distress Syndrome Address Alveolar Macrophages Antibodies Automobile Driving Binding Biological Body Weight Cell Survival Cell physiology Cells Cessation of life Clinical Data Dependence Disease Dose Enterobacteria phage P1 Cre recombinase Extracellular Signal Regulated Kinases Gene Expression Gene Expression Profile Genetic Polymorphism Human IFNAR1 gene Immune system Impairment In Vitro Inflammation Inflammatory Inflammatory Response Injury Interferon Type I Interleukin-4 KRP protein Leukocytes Lung MAP2K1 gene MAPK3 gene MEKs Macrophage Macrophage Activation Measures Mitogen-Activated Protein Kinases Modeling Mus Myelogenous Myeloid Cells Neutrophil Infiltration Neutrophilia Outcome Pathway interactions Patients Phenotype Phosphorylation Phosphotransferases Process Protein Isoforms Protein Kinase Publishing Pulmonary Inflammation Recovery Regulation Regulatory Pathway Resolution Rodent Role Route SARS-CoV-2 infection Signal Pathway Signal Transduction Site Source Sterility Testing Therapeutic Time Toxic effect Translating Wild Type Mouse Work epithelial repair experience extracellular improved injured injury recovery lung injury mortality mouse model mutant new therapeutic target novel novel therapeutic intervention pre-clinical repaired response single-cell RNA sequencing wound healing

项目摘要

ABSTRACT With the surge of ARDS cases associated with SARS-CoV-2 infection, there is an urgent need to understand novel pathways involved in resolution of lung inflammation and injury to provide the basis for new therapeutic approaches. Studies comparing rodent and human lung injury gene expression signatures reveal conserved pathways, including MAPK/ERK activation during injury. More recently, we found a genetic polymorphism in MAP2K2 associates with death in ARDS, suggesting a biological role in ALI recovery. We demonstrated that MAP2K1/MAP2K2 (MEK1/MEK2) activation in macrophages promotes pro-inflammatory pathways and inhibits reparative ones, making it a potential target to manipulate macrophage phenotypes in ALI. In preclinical acute lung injury (ALI) models, inhibition of MEK1/MEK2 improves measures of ALI, including faster recovery of body weight, reduced pulmonary neutrophilia, and greater reparative macrophage activation. These results suggest that MEK pathways could be effective targets in ALI. To address the isoform and cell source driving this improvement in outcome, we generated mice deficient in MEK1 in myeloid cells (LysmCre+Mek1fl). These mice have no phenotype in naïve conditions, but experience 100% mortality with LPS-induced ALI using a moderate LPS dose from which all wild-type mice recover. These mice have a similar early inflammatory response to LPS but fail to turn off inflammation at later time-points. This phenotype can be completely rescued with IFNAR1 blockade, suggesting MEK1 suppression of type I interferon responses is critical for ALI resolution. We also found sustained MEK2/p-ERK activation in the absence of MEK1, suggesting that MEK1 is critical for MEK2 deactivation to promote ALI resolution. In support of this hypothesis, we found that mice lacking MEK2 globally or in the leukocytes compartment have faster ALI resolution. The proposed aims below outline our approach to identify how MEK isoforms work in concert to regulate myeloid cell responses to better define and target a novel regulatory pathway in ALI. In aim 1, we will determine MEK1 and MEK2 cell-specific roles and signaling pathways in ALI and test our will test our hypothesis that MEK1 is required to deactivate MEK2 in alveolar macrophages to promote resolution of lung inflammation. In aim 2, we plan to evaluate MEK1 and MEK2 interactions and mechanisms of MEK2 deactivation. We will use MEK1 mutants to test our hypothesis that MEK2 is deactivated by binding to pT292 MEK1, and we will determine how these mutants alter macrophage inflammatory (LPS) and reparative (IL-4) responses. In aim 3, we plan to test MEK1 and MEK2 degraders as therapeutics in murine models of ALI. We will test our hypothesis that MEK2-specific degradation will result in faster ALI resolution. These studies will advance our understanding of how the immune system stops inflammation and promotes ALI resolution, revealing new therapeutic targets and approaches that could be brought to the bedside.

抽象的随着与 SARS-CoV-2 感染相关的 ARDS 病例激增，迫切需要了解参与解决肺部炎症和损伤的新途径为新疗法提供基础比较啮齿动物和人类肺损伤基因表达特征的研究揭示了保守性。最近，我们发现了一种基因多态性。 MAP2K2 与 ARDS 死亡相关，表明其在 ALI 恢复中具有生物学作用。巨噬细胞中的 MAP2K1/MAP2K2 (MEK1/MEK2) 激活促进促炎途径并抑制修复性的，使其成为在临床前急性急性肺损伤中操纵巨噬细胞表型的潜在目标。肺损伤 (ALI) 模型，抑制 MEK1/MEK2 可以改善 ALI 的指标，包括身体更快恢复这些结果表明，体重增加、肺中性粒细胞减少和修复性巨噬细胞活化增强。 MEK 途径可能是 ALI 的有效靶标，以解决驱动这一问题的异构体和细胞来源。为了改善结果，我们培育了骨髓细胞中缺乏 MEK1 的小鼠 (LysmCre+Mek1fl)。在幼稚条件下没有表型，但使用中等强度的 LPS 诱导的 ALI 死亡率为 100% 所有野生型小鼠恢复的 LPS 剂量这些小鼠对 LPS 具有相似的早期炎症反应。但在以后的时间点无法消除炎症，这种表型可以用 IFNAR1 完全挽救。阻断，表明 MEK1 抑制 I 型干扰素反应对于 ALI 的解决至关重要。发现在没有 MEK1 的情况下持续的 MEK2/p-ERK 激活，表明 MEK1 对于 MEK2 至关重要为了支持这一假设，我们发现小鼠体内普遍缺乏 MEK2。或在白细胞区室中具有更快的 ALI 分辨率。下面提出的目标概述了我们的方法。确定 MEK 亚型如何协同作用来调节骨髓细胞反应，以更好地定义和靶向新型药物在目标 1 中，我们将确定 MEK1 和 MEK2 细胞特异性作用和信号传导。 ALI 中的通路并测试我们的意志测试我们的假设，即需要 MEK1 才能使肺泡中的 MEK2 失活巨噬细胞促进肺部炎症消退在目标 2 中，我们计划评估 MEK1 和 MEK2。我们将使用 MEK1 突变体来检验我们的假设。 MEK2 通过与 pT292 MEK1 结合而失活，我们将确定这些突变体如何改变巨噬细胞在目标 3 中，我们计划测试 MEK1 和 MEK2 降解剂。我们将测试我们的假设，即 MEK2 特异性降解会导致 ALI 小鼠模型的治疗。这些研究将促进我们对免疫系统如何停止的理解。炎症并促进 ALI 消退，揭示新的治疗靶点和方法带到了床边。