Document text
Principal Investigator: Anne M. Manicone
Organization: UNIVERSITY OF WASHINGTON
Fiscal Year: 2024
Award: $717,227
Funding agency: National Heart Lung and Blood Institute
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.
Terms: <ARDS><ATP-protein phosphotransferase><Acceleration><Acute Lung Injury><Acute Pulmonary Injury><Acute Respiratory Distress><Acute Respiratory Distress Syndrome><Address><Adult ARDS><Adult RDS><Adult Respiratory Distress Syndrome><Alveolar Macrophages><Antibodies><Automobile Driving><B cell growth factor><B-Cell Differentiation Factor-1><B-Cell Growth Factor-1><B-Cell Growth Factor-I><B-Cell Proliferating Factor><B-Cell Stimulating Factor><B-Cell Stimulating Factor-1><B-Cell Stimulation Factor-1><B-Cell Stimulatory Factor-1><BCDF-1><BCGF><BCGF-1><BCSF 1><BSF-1><BSF1><Binding><Binetrakin><Biological><Blood leukocyte><Body Weight><COVID-19 infection><COVID-19 virus infection><COVID19 infection><CRE Recombinase><Cell Body><Cell Communication and Signaling><Cell Function><Cell Physiology><Cell Process><Cell Signaling><Cell Survival><Cell Viability><Cells><Cellular Function><Cellular Physiology><Cellular Process><Cessation of life><Clinical><Da Nang Lung><Data><Death><Dependence><Disease><Disorder><Dose><ERK 1><ERK MAP Kinases><ERK1><ERK1 Kinase><Enterobacteria phage P1 Cre recombinase><Expression Signature><Extracellular Signal Regulated Kinases><Extracellular Signal-Regulated Kinase 1><Extracellular Signal-Regulated MAP Kinases><Gene Expression><Gene Expression Profile><Genetic Polymorphism><Human><IFNAR><IFNAR1><IFNAR1 gene><IL-4><IL4 Protein><Immune system><Impairment><In Vitro><Inflammation><Inflammatory><Inflammatory Response><Injury><Interferon Type I><Interleukin-4><Interleukin-4 Precursor><Intracellular Communication and Signaling><Isoforms><KRP protein><Kinase Family Gene><Kinases><Leukocytes><Leukocytes Reticuloendothelial System><Lung><Lung Inflammation><Lung Respiratory System><Lung damage><Lymphocyte Stimulatory Factor 1><MAP Kinase 3><MAP Kinase Kinase 1><MAP kinase><MAP2K1><MAP2K1 gene><MAPK ERK Kinases><MAPK/ERK Kinase 1><MAPK3><MAPK3 Mitogen-Activated Protein Kinase><MAPK3 gene><MAPKK1><MCGF-2><MEK-1><MEK1><MEKs><MKK1><Macrophage><Macrophage Activation><Marrow leukocyte><Mast Cell Growth Factor-2><Measures><Mice><Mice Mammals><Mitogen-Activated Protein Kinase 3><Mitogen-Activated Protein Kinase 3 Gene><Mitogen-Activated Protein Kinase Kinase-1><Mitogen-Activated Protein Kinases><Modeling><Modern Man><Molecular Interaction><Murine><Mus><Myelogenous><Myeloid><Myeloid Cells><Mφ><Neutrophil Infiltration><Neutrophil Recruitment><Neutrophilia><Neutrophilic Infiltrate><Outcome><P44ERK1><PRKMK1><PSTkinase p44mpk><Pathway interactions><Patients><Phenotype><Phosphorylation><Phosphotransferase Gene><Phosphotransferases><Pneumonitis><Process><Protein Isoforms><Protein Kinase><Protein Phosphorylation><Publishing><Pulmonary Inflammation><Pulmonary Macrophages><Recovery><Regulation><Regulatory Pathway><Resolution><Rodent><Rodentia><Rodents Mammals><Role><Route><SARS-CoV-2 infection><SARS-CoV2 infection><Severe acute respiratory syndrome coronavirus 2 infection><Shock Lung><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Site><Source><SpKRP 85><SpKRP 95><Sterility><Stiff lung><Subcellular Process><T-Cell Growth Factor 2><Testing><Therapeutic><Threonine/Tyrosine Protein Kinase><Time><Toxic effect><Toxicities><Translating><Transphosphorylases><White Blood Cells><White Cell><Wild Type Mouse><Work><Wound Repair><bacteriophage P1 recombinase Cre><biologic><biological signal transduction><coronavirus disease 2019 infection><driving><epithelial repair><experience><extracellular><extracellular signal related kinase><gene expression pattern><gene expression signature><glycogen synthase a kinase><hydroxyalkyl protein kinase><improved><infected with COVID-19><infected with COVID19><infected with SARS-CoV-2><infected with SARS-CoV2><infected with coronavirus disease 2019><infected with severe acute respiratory syndrome coronavirus 2><injured><injuries><injury recovery><kinase-related protein><kinesin II><lung injury><mortality><mouse model><murine model><mutant><new drug target><new druggable target><new pharmacotherapy target><new therapeutic approach><new therapeutic intervention><new therapeutic strategies><new therapeutic target><new therapy approaches><new therapy target><new treatment approach><new treatment strategy><novel><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic approach><novel therapeutic intervention><novel therapeutic strategies><novel therapeutic target><novel therapy approach><novel therapy target><p44 MAPK><pathway><phosphorylase b kinase kinase><polymorphism><pre-clinical><preclinical><pulmonary><pulmonary damage><pulmonary injury><pulmonary tissue damage><pulmonary tissue injury><recovery after injury><recovery following injury><recovery post injury><repair><repaired><resolutions><response><scRNA-seq><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><social role><sterile><transcriptional profile><transcriptional signature><wet lung><white blood cell><white blood corpuscle><wildtype mouse><wound healing><wound recovery><wound resolution>