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.

摘要 随着与SARS-CoV-2感染相关的ARDS病例的激增，迫切需要了解 参与肺部炎症和损伤消退的新途径，为新的治疗方法提供基础。 接近。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的 MAPK/ERK信号通路，包括损伤时的MAPK/ERK激活。最近，我们发现了一种遗传多态性， MAP 2K 2与ARDS死亡相关，提示其在ALI恢复中的生物学作用。我们证明了 巨噬细胞中的MAP 2K 1/MAP 2K 2（MEK 1/MEK 2）活化促进促炎途径并抑制 修复的，使其成为一个潜在的目标，操纵巨噬细胞表型在ALI。临床前急性 在肺损伤（ALI）模型中，抑制MEK 1/MEK 2改善了ALI的测量，包括更快的身体恢复 体重、减少的肺嗜中性粒细胞和更大的修复性巨噬细胞活化。这些结果表明 MEK通路可能是ALI的有效靶点。为了解决驱动这一点的同种型和细胞来源， 为了改善结果，我们产生了骨髓细胞中MEK 1缺陷的小鼠（LysmCre+ Mek 1fl）。这些小鼠 在初始条件下没有表型，但使用中度 所有野生型小鼠从中恢复的LPS剂量。这些小鼠对LPS有类似的早期炎症反应 但在以后的时间点不能关闭炎症。这种表型可以用IFNAR 1完全挽救 阻断，表明MEK 1抑制I型干扰素应答对于ALI消退至关重要。我们也 发现在不存在MEK 1的情况下持续的MEK 2/p-ERK激活，表明MEK 1对MEK 2至关重要。 去激活以促进ALI的解决。为了支持这一假设，我们发现缺乏MEK 2的小鼠在全球范围内 或在白细胞隔室中具有更快的ALI消退。以下拟议目标概述了我们的方法， 确定MEK同种型如何协同调节骨髓细胞反应，以更好地定义和靶向新的 ALI中的调节通路。在目标1中，我们将确定MEK 1和MEK 2细胞特异性作用和信号传导 我们的假设是，肺泡上皮细胞中MEK 2的失活需要MEK 1的参与， 巨噬细胞，以促进肺部炎症的解决。在目标2中，我们计划评估MEK 1和MEK 2 MEK 2失活的相互作用和机制。我们将使用MEK 1突变体来验证我们的假设， MEK 2通过与pT 292 MEK 1结合而失活，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。在目标3中，我们计划测试MEK 1和MEK 2降解剂， 在ALI的鼠模型中的治疗剂。我们将测试我们的假设，即MEK 2特异性降解将导致 更快的ALI解决方案。这些研究将推进我们对免疫系统如何停止的理解 炎症和促进ALI的解决，揭示了新的治疗靶点和方法， 带到床边。