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

• 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
• 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

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 涉及肺部感染和损伤的新型途径，为新疗法提供基础 方法。比较啮齿动物和人肺损伤基因表达特征的研究表明保守 途径，包括受伤期间MAPK/ERK激活。最近，我们在 MAP2K2与ARDS中的死亡相关联，表明在ALI恢复中起着生物学作用。我们证明了这一点 MAP2K1/MAP2K2（MEK1/MEK2）巨噬细胞激活促进促炎途径并抑制 修复性的，使其成为操纵巨噬细胞表型的潜在目标。在临床前急性 肺损伤（ALI）模型，MEK1/MEK2的抑制可改善ALI的测量值，包括更快的身体恢复 重量，肺中性粒细胞减少和更大的修复巨噬细胞激活。这些结果表明 MEK途径可能是ALI的有效目标。解决驱动此的同工型和细胞来源 结果的改善，我们产生了髓样细胞中MEK1缺乏的小鼠（Lysmcre+MEK1FL）。这些老鼠 在幼稚的条件下没有表型，但是使用现代的LPS诱导的ALI经历了100％死亡率 所有野生型小鼠恢复的LPS剂量。这些小鼠对LP具有相似的早期炎症反应 但是无法在以后的时间点关闭炎症。可以使用IFNAR1完全检索此表型 封锁，表明MEK1抑制I型干扰素反应对于ALI分辨率至关重要。我们也是 发现在没有MEK1的情况下持续的MEK2/P-ERK激活，这表明MEK1对MEK2至关重要 停用以促进ALI分辨率。为了支持这一假设，我们发现全球缺乏MEK2的小鼠 或在白细胞中具有更快的ALI分辨率。拟议的目的下面概述了我们的方法 确定MEK同工型如何协同调节髓样细胞反应以更好地定义和针对新颖 阿里的监管途径。在AIM 1中，我们将确定MEK1和MEK2细胞特异性角色和信号传导 ALI中的途径并测试我们的意志，测试我们的假设，即MEK1需要在肺泡中停用MEK2 巨噬细胞促进肺注射的分辨率。在AIM 2中，我们计划评估MEK1和MEK2 MEK2停用的相互作用和机制。我们将使用MEK1突变体来检验我们的假设 MEK2通过与PT292 MEK1结合而停用，我们将确定这些突变体如何改变巨噬细胞 炎症（LPS）和修复（IL-4）反应。在AIM 3中，我们计划测试MEK1和MEK2降解器 阿里鼠模型中的治疗剂。我们将测试我们的假设，即MEK2特异性降解将导致 更快的ALI分辨率。这些研究将提高我们对免疫系统如何停止的理解 炎症并促进ALI分辨率，揭示了可能是新的治疗靶标和方法 买到床边。