Cystic fibrosis (CF) is a genetic disease caused by mutations in the cystic fibrosis transmembrane regulator (CFTR) gene. CF airways are immunocompromised and become colonized with bacteria soon after birth. Chronic bacterial infection leads to persistent and severe neutrophil-dominated pulmonary inflammation, high lung protease levels, lung damage and a decline in FEV1. CFTR modulator/correctors from Vertex such as TRIKAFTA significantly increase CF patient lung function by >10% but do not bring it into the normal range and for patients with pre-existing bacterial lung infections, do not clear bacteria from their lungs. Moreover, these compounds do not treat CF patients with nonsense mutations where no CFTR protein is produced. Thus, there is a critical unmet need for novel, CFTR mutation-agnostic therapies to help clear bacteria from CF lungs and limit neutrophilic lung damage. Short Palate LUng and Nasal epithelial Clone 1 (SPLUNC1) is a secreted protein that is highly expressed in the lung, where it plays a key role in maintaining lung health. SPLUNC1 is a CF gene modifier, and patients with reduced SPLUNC1 levels have lower FEV1 and exacerbate more frequently. Orai1 is a ubiquitously expressed plasma membrane Ca2+ channel that regulates inflammation. We found that SPLUNC1 inhibits Orai1. However, SPLUNC1 is rapidly degraded by neutrophil elastase (NE), which we posit results in greater Orai1 activity and more inflammation. Consistent with this, our preliminary data indicate that Orai1 is upregulated in CF patient lung immune cells. Given Orai1's proximal role in the immune response, Orai1 is thus an attractive target whose inhibition is predicted to help resolve CF inflammation. Eldec Pharma has developed a robust, novel peptidomimetic called ELD607, which reprises SPLUNC1's ability to inhibit Orai1, yet is significantly more stable in the presence of NE, and significantly more potent/efficacious. ELD607 is stable in proteolytic CF sputum, and inhibits Ca2+-influx in freshly-isolated CF patient peripheral neutrophils and in CF sputum-derived immune cells in a mutation-agnostic fashion. In murine lung infection models with common CF pathogens including P. aeruginosa and S. aureus, a single, inhaled dose of ELD607 reduced lung inflammation (neutrophilia, cytokines, NE) by 90%, decreased lung bacterial infection by 3-4 log10 CFUs and increased survival. In a chronic CF model (SCNN1B mice), ELD607 reduced neutrophilia and increased survival. These experiments demonstrate that rebalancing the lung's inflammatory response by inhibiting Orai1 enhances the lungs' natural ability to clear pathogens. In this proposal, we will use wild-type and CF mice to understand which immune effector cells are regulated by ELD607. The CF ferret model developed by Dr Engelhardt and coworkers at the University of Iowa spontaneously develops chronic CF lung disease including inflammation and bacterial infection. We will first validate that we can deliver sufficient doses of ELD607 via nebulizer to safely inhibit Orai1 in wild-type and CF ferret lungs. Then we will chronically administer ELD607 to CF ferrets with existing lung disease in order to study the impact of ELD607 on CF disease progression.
Public Health Relevance Statement: Public Health Narrative Due to a genetically induced imbalance in salt and water transport in cystic fibrosis (CF) lungs, mucus is dehydrated and the lungs fail to eradicate bacteria, leaving the airways prone to chronic infection and inflammation that ultimately destroy the lung. In this application, we propose to develop peptide-based therapies that can shift the lung's immune balance to efficiently clear bacteria from the lung.
Project Terms: Aerosols; Affect; Bacteria; Bacterial Infections; bacteria infection; bacterial disease; Birth; Parturition; Capital; Cell membrane; Cytoplasmic Membrane; Plasma Membrane; plasmalemma; Cells; Cell Body; Cystic Fibrosis; Mucoviscidosis; Equilibrium; balance; balance function; Ferrets; Foundations; Genes; Regulator Genes; Transcriptional Regulatory Elements; regulatory gene; trans acting element; Haemophilus influenzae; H influenzae; H. influenzae; Inflammation; Iowa; Lung; Lung Respiratory System; pulmonary; Lung diseases; Pulmonary Diseases; Pulmonary Disorder; disease of the lung; disorder of the lung; lung disorder; Biological Models; Biologic Models; Model System; Mucous body substance; Mucus; mucous; Mus; Mice; Mice Mammals; Murine; Mutation; Genetic Alteration; Genetic Change; Genetic defect; genome mutation; Nebulizer; Blood Neutrophil; Blood Polymorphonuclear Neutrophil; Marrow Neutrophil; Neutrophilic Granulocyte; Neutrophilic Leukocyte; Polymorphonuclear Cell; Polymorphonuclear Leukocytes; Polymorphonuclear Neutrophils; neutrophil; Patients; Esteroproteases; Peptidases; Protease Gene; Proteases; Proteinases; Proteolytic Enzymes; Peptide Hydrolases; Peptides; Pharmacokinetics; Drug Kinetics; Play; Privatization; Proteins; Chrysemonas; Flavimonas; Pseudomonas; P aeruginosa; P. aeruginosa; Pseudomonas pyocyanea; Pseudomonas aeruginosa; Public Health; Reagent; social role; Role; salt; Sodium Chloride; S aureus; S. aureus; Staph aureus; Staphylococcus aureus; Testing; Translating; Universities; Water; Hydrogen Oxide; Cystic Fibrosis Transmembrane Conductance Regulator; CFTR; CFTR Protein; cystic fibrosis transmembrane regulator; Leukocyte Elastase; Granulocyte Elastase; Lysosomal Elastase; Neutrophil Elastase; PMN Elastase; Polymorphonuclear Leukocyte Elastase; cytokine; Alveolar Macrophages; Pulmonary Macrophages; Immunocompromised Host; Immunocompromised; Immunocompromised Patient; Immunosuppressed Host; immunosuppressed patient; Burkholderia cepacia; B cenocepacia; B cepacia; B. cenocepacia; B. cepacia; Burkholderia cenocepacia; P cepacia; P. cepacia; Pseudomonas cepacia; Mediating; Normal Range; Normal Values; Neutrophilia; Immunology; base; improved; Peripheral; Chronic; lung function; pulmonary function; Disease Progression; Lung damage; pulmonary damage; pulmonary injury; pulmonary tissue damage; pulmonary tissue injury; lung injury; Immunological response; host response; immune system response; immunoresponse; Immune response; Therapeutic; Effector Cell; programs; CF lung disease; cystic fibrosis lung; cystic fibrosis lung disease; Pulmonary Cystic Fibrosis; Immunes; Immune; cell type; System; non-sense mutation; Nonsense Mutation; Nasal Epithelium; novel; Palate; FEV1; FEV1%VC; Forced Expiratory Volume 1 Test; Forced Expiratory Volume in 1 Second; PFT/FEV1; Pulmonary Function Test/Forced Expiratory Volume 1; Modeling; Bacterial Antibiotic Resistance; antibiotic resistant bacteria; bacterial antibiotic resistant; bacterial resistance to antibiotic; Burkholderia cepacia complex; B cepacia complex; B. cepacia complex; Inflammatory Response; MRSA; Methicillin Resistant S Aureus; Methicillin Resistant S. Aureus; methicillin-resistant S. aureus; methicillin resistant Staphylococcus aureus; preventing; prevent; Dose; Data; Eldec; Ero Test; Estratest; Malogen; Metandren; Neohombreol M; Orchisterone-M; Oreton methyl; Testomet; Testotonic B; Testovis; Testred; Glosso-Sterandryl; Antimicrobial Effect; anti-microbial effect; Immunomodulators; IMiD; Immune modulatory therapeutic; immune modulating agents; immune modulating drug; immune modulating therapeutics; immune modulators; immune modulatory agents; immune modulatory drugs; immunomodulating agents; immunomodulatory agents; immunomodulatory drugs; immunomodulatory therapeutics; novel strategies; new approaches; novel approaches; novel strategy; pathogen; cystic fibrosis airway; CF airway; cystic fibrosis mouse; CF mice; CF mouse model; cystic fibrosis mouse model; cystic fibrosis patients; CF patients; individuals with CF; individuals with cystic fibrosis; patients with CF; patients with cystic fibrosis; Population; antimicrobial; anti-microbial; peptidomimetics; peptide mimetic; peptide mimic; aerosolized; disease-causing mutation; combat; Regimen; Pulmonary Inflammation; Lung Inflammation; Pneumonitis; multidrug-resistant Pseudomonas aeruginosa; MDR P aeruginosa; MDR P. aeruginosa; MDR Pseudomonas aeruginosa; multi-drug resistant P. aeruginosa; multi-drug resistant Pseudomonas aeruginosa; multidrug resistant P. aeruginosa; multidrug-resistant P. aeruginosa; experimental study; experiment; experimental research; clinical development; Inhalation; Inhaling; Genetic Diseases; genetic condition; genetic disorder; chronic infection; persistent infection; Lung infections; pulmonary infections; Cystic Fibrosis sputum; CF sputum; cystic fibrosis infection; CF infection; infection in CF; infection in cystic fibrosis; lung health; pulmonary health
