The Epithelial Na+ Channel (ENaC) is the rate limiting step for Na+ absorption across many epithelia. In the airways of cystic fibrosis (CF) patients, ENaC is abnormally hyperactive leading to disproportionate Na+ absorption and a depletion of airway surface liquid (ASL) volume. ENaC hyperactivity contributes to mucus stasis and increased incidence of airway infections that frequently lead to the death of the patient. Despite previous efforts, there are no existing therapies to treat abnormal ENaC activity in the lung. Using a proteomic screen, we have identified the protein SPLUNC1 as a potent allosteric inhibitor of ENaC that binds extra-cellularly to ENaC, reducing the amount of ENaC in the plasma membrane and subsequently limiting ENaC activity. We have also identified the ENaC-inhibitory domain of SPLUNC1, and have synthesized small peptide that corresponds to this domain, called SPX-101. This peptide robustly inhibits ENaC and prevents ASL hyperabsorption in CF airway cultures for 24 h following a single dose. SPX-101 continues to function in the presence of neutrophil elastase, which is highly abundant in CF airways, suggesting that this peptide may be therapeutically beneficial in the treatment of CF lung disease. We have also demonstrated that SPX-101 improves survival in a transgenic mouse model of CF that specifically overexpresses ENaC in the lung epithelia (?ENaC-Tg) and that SPX-101 can be effectively delivered to the lung via nebulization. Notably, SPX-101 does not induce diuresis or hyperkalemia when delivered by intravenous infusion in rats. Furthermore, nebulized SPX-101 remains sequestered within the lung and
Public Health Relevance Statement: The epithelial sodium channel (ENaC) is a molecular target for cystic fibrosis (CF). When the cystic fibrosis transmembrane conductance regulator protein is inactive due to genetic mutation, the body loses the ability to secrete chloride ions into the fluid that lines the airways. Since salt in this fluid controls the total volume of the fluid decreases and the mucus in the airways becomes dehydrated. The loss of salt due to reduced chloride secretion is made worse by ENaC, which removes sodium from the airway fluid and further dehydrates the lung. ENaC inhibition can counteract the loss of fluid and accumulation of viscous, dehydrated mucus that blocks small airways and provides a nutrient source for bacterial growth. We have identified a novel peptide agent that inhibits ENaC activity and may provide a valuable new approach for treatment. We propose to test this novel peptide, known as SPX-101, for the treatment of CF.
Project Terms: absorption; Address; Adult; Advanced Development; Adverse effects; Age-Years; Agreement; Air; airway surface liquid; Amiloride; Animal Model; Bacteria; base; Binding; Biological Assay; Biological Sciences; Blood Circulation; bronchial epithelium; Canis familiaris; Cell membrane; Cells; Cessation of life; Chloride Ion; Chlorides; Chronic; Chronic lung disease; Climacteric; Clinic; Clinical; clinical application; Clinical Trials; cost; Cystic Fibrosis; cystic fibrosis airway; cystic fibrosis mouse; cystic fibrosis patients; Cystic Fibrosis Transmembrane Conductance Regulator; Dehydration; Disease; disease-causing mutation; Diuresis; DNA Sequence Alteration; Dose; Epithelial; Epithelial Cells; epithelial Na+ channel; Epithelium; Exposure to; extracellular; Functional disorder; Goals; Grant; Growth; Half-Life; Health; Height; Human; Hydration status; Hyperactive behavior; hyperkalemia; improved; Incidence; Infection; inhibitor/antagonist; Intravenous infusion procedures; Kidney; Lead; Leukocyte Elastase; Life Expectancy; Liquid substance; Lung; Measures; Microscopic; Modeling; Molecular Target; Mucolytics; Mucous body substance; Mutation; Nebulizer; novel; novel strategies; novel therapeutics; Nutrient; Organ; overexpression; palliative; patient subsets; Patients; peptide drug; Peptides; Pharmaceutical Preparations; Pharmacology; Phase; Pilot Projects; Plasma; prevent; Proteins; Proteomics; Pulmonary Cystic Fibrosis; Quality of life; Rattus; Regulation; Saline; Sheep; Small Business Innovation Research Grant; small molecule; Sodium; Sodium Chloride; Source; success; Surface; Testing; Therapeutic; Tissues; Toxic effect; Toxicology; Transgenic Mice; Translating; translational study; Validation; water diffusion; Work
