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Principal Investigator: Franco Rafael D'Alessio
Organization: JOHNS HOPKINS UNIVERSITY
Fiscal Year: 2024
Award: $787,156
Funding agency: National Heart Lung and Blood Institute
Pneumonia (PNA) is one of the leading causes of death worldwide. PNA can result in devastating acute
inflammatory injury in the lung manifesting in acute respiratory distress syndrome (ARDS). Current treatments for
PNA have focused on the pathogens, but do not target excessive lung inflammation elicited by the host immune
response. Both the emergence of new infections, typified by COVID-19, and the expanding impact of antimicrobial
resistant pathogens, highlight the limitations of our current armamentarium and underscore the need to identify
additional therapeutic targets in PNA-induced ARDS. With the understanding that resolution of PNA is an
actively regulated program to promote return to homeostasis, our work has focused on identifying cellular and
molecular mediators of this resolution phase. Others and we have demonstrated that regulatory T cells (Tregs)
promote resolution of infectious-ARDS.
Our strong preliminary data has identified lung-derived Treg DHX58, which encodes an RNA helicase protein
essential for antiviral responses, as a candidate gene upregulated during the resolution phase of ARDS. DHX58-
deficient animals fail to resolve lung inflammation after Streptococcus pneumoniae-ARDS with significantly
diminished lung Treg numbers during injury resolution, implicating DHX58 in optimal Treg function in vivo. Further,
we observed significantly increased 30-day mortality among carriers of a putative loss-of-function variant of DHX58
with infectious ARDS (71% vs. 47%, p=0.01), underscoring the potential clinical impact of DHX58 in ARDS
outcomes. Our in-silico analysis of the DHX58 promoter identified numerous estrogen responsive elements (ERE).
Indeed, DHX58 expression was induced in Tregs by estradiol (E2). Importantly, our published work showed that
therapeutic E2 promotes resolution of preclinical PNA-ARDS in a Treg-dependent manner. Estrogen receptor beta
(ER) was necessary for both Treg-dependent rescue of lymphopenic hosts and Treg-mediated suppression of
pro-inflammatory cytokine production in macrophages in vitro. Preliminary gene expression analysis and high-
dimensional flow cytometry implicate E2 and its downstream-target, DHX58, in the regulation of critical Treg
transcription factors (TFs), notably Foxp3 and GATA3. Thus, we hypothesize that E2, in part via ER-dependent
upregulation of DHX58, orchestrates critical Treg pro-resolution functions, through regulating expression of key
TFs in Tregs. The goals of this proposal are to determine the cellular, molecular and transcriptional determinants
of E2-ER-DHX58 in Treg-mediated resolution of PNA-ARDS to provide the mechanistic underpinnings of the
regulation and functional role of ER in Tregs.
Terms: <ARDS><Acute><Acute Pneumonia><Acute Respiratory Distress><Acute Respiratory Distress Syndrome><Adult ARDS><Adult RDS><Adult Respiratory Distress Syndrome><Agonist><Alveolar><Animals><Anti-viral Response><Aquadiol><Autoregulation><Basal Transcription Factor><Basal transcription factor genes><Blood Neutrophil><Blood Polymorphonuclear Neutrophil><COVID-19><CV-19><Candidate Disease Gene><Candidate Gene><Cause of Death><Cell Body><Cell Communication and Signaling><Cell Count><Cell Function><Cell Number><Cell Physiology><Cell Process><Cell Signaling><Cells><Cellular Function><Cellular Physiology><Cellular Process><Cellular biology><Clinical><Clinical Trials><Coronavirus Infectious Disease 2019><Critical Paths><Critical Pathways><D pneumoniae><D. pneumoniae><Da Nang Lung><Data><Development><Dimenformon><Diogyn><Diogynets><Diplococcus pneumoniae><ER-BETA><ERbeta><ERβ><ESR-BETA><ESR2><ESR2 gene><ESRB><ESTRB><Elements><Enhancer-Binding Protein GATA3><Estrace><Estradiol><Estradiol-17 beta><Estradiol-17beta><Estraldine><Estrogen Receptor 2><Estrogen Receptor beta><Estrogen Receptor β><Estrogenic Agents><Estrogenic Compounds><Estrogens><FOXP3><FOXP3 gene><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Forkhead Box P3><GATA-3 factors><GATA-3 protein><GATA-Binding Protein 3><GATA3><GATA3 gene><GATA3 protein><GATA3 transcription factor><Gene Expression><Gene Expression Monitoring><Gene Expression Pattern Analysis><Gene Expression Profiling><Gene Transcription><General Transcription Factor Gene><General Transcription Factors><Genetic Transcription><Goals><Homeostasis><Human><Immune><Immune response><Immunes><Immunological response><In Vitro><Infection><Inflammatory><Inflammatory Response><Injury><Intracellular Communication and Signaling><JM2><Lung><Lung Alveolar Epithelia><Lung Inflammation><Lung Respiratory System><Lung damage><Macrophage><Marrow Neutrophil><Mediating><Mediator><Mice><Mice Mammals><Modeling><Modern Man><Molecular><Murine><Mus><Mφ><NR3A2><Neutrophilic Granulocyte><Neutrophilic Leukocyte><Outcome><Ovocyclin><Ovocylin><Pathway interactions><Phase><Physiological Homeostasis><Pilot Projects><Play><Pneumococcus><Pneumonia><Pneumonitis><Polymorphonuclear Cell><Polymorphonuclear Leukocytes><Polymorphonuclear Neutrophils><Pre-Clinical Model><Preclinical Models><Production><Progynon><Proteins><Publishing><Pulmonary Inflammation><RNA Expression><RNA Helicase><Regulation><Regulatory T-Lymphocyte><Resolution><Role><S pneumoniae><S. pneumoniae><SCURFIN><Shock Lung><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Stiff lung><Streptococcus pneumoniae><Subcellular Process><Testing><Therapeutic><Therapeutic Effect><Therapeutic Estradiol><Therapeutic Estrogen><Transcript Expression Analyses><Transcript Expression Analysis><Transcription><Transcription Factor Proto-Oncogene><Transcription factor genes><Treg><Up-Regulation><Upregulation><Variant><Variation><Work><acute infection><alveolar epithelium><analyze gene expression><anti-microbial resistant pathogen><antibiotic resistant pathogen><antimicrobial resistant pathogen><biological signal transduction><cell biology><coronavirus disease 2019><coronavirus disease-19><coronavirus infectious disease-19><cytokine><developmental><disease causing variant><disease model><disease-causing mutation><disorder model><drug resistant pathogen><estrogenic><flow cytophotometry><gene expression analysis><gene expression assay><high dimensionality><host response><immune system response><immunoresponse><in silico><in vitro activity><in vivo><inflammatory lung disease><injuries><loss of function><lung injury><mortality><neutrophil><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><next generation therapeutics><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutics><novel therapy><pathogen><pathogenic variant><pathway><pilot study><pneumonia model><pneumonia models><pneumonia therapy><pneumonia treatment><pre-clinical><preclinical><programs><promoter><promotor><pulmonary><pulmonary damage><pulmonary injury><pulmonary tissue damage><pulmonary tissue injury><regulatory T-cells><repair><repaired><resolutions><response><social role><therapeutic evaluation><therapeutic target><therapeutic testing><transcription factor><transcriptional profiling><treat pneumonia><wet lung>