The Role of DNA and Actin Polymers on the Polymer Structure and Rheology of Cystic Fibrosis Sputum and Depolymerization by Gelsolin or Thymosin Beta 4

Abstract

Mucus clearance is the first line of pulmonary defense against inhaled irritants, microorganisms, and allergens. In health, the gel-forming mucins are the principal polymeric components of airway mucus but in cystic fibrosis (CF), the necrotic death of inflammatory and epithelial cells releases a network of copolymerized extracellular DNA and filamentous (F-) actin-producing secretions that are similar to pus and difficult to clear by cilia or airflow. The large amounts of F-actin in CF sputum suggested that thymosin beta4 (Tbeta4) or gelsolin could depolymerize the secondary polymer network of CF sputum. Dose-dependent CF sputum rheology and polymer structure were measured before and after the addition of excipient, dornase alfa, Tbeta4, gelsolin, and Tbeta4 or gelsolin with dornase for 30 min. Sputum was also incubated with Tbeta4 30 microg/mL, gelsolin 10 microg/mL or excipient for 0, 5, 10, 15, 20, or 60 min. There was a dose-dependent decrease in cohesivity with Tbeta4 and a time-dependent decrease in cohesivity at 30 microg/mL. With the combination of dornase alfa and Tbeta4 at 1.5 microg/mL, there was a 65% decrease in elasticity (P = 0.013). There was a time-dependent decrease in cohesivity (P = 0.0004) and elasticity (P = 0.047) with gelsolin and a dose-dependent fall in cohesivity (P = 0.0008). An apparent synergy of Tbeta4 or gelsolin on actin and dornase on DNA may be explained by the combined effect of actin depolymerization and DNA filament severing or by virtue of actin depolymerization increasing the effectiveness of dornase alfa.