Mobility and stability of gene complexes in biogels

Abstract

The tenacious secretions lining the conductive airways of cystic fibrosis (CF) patients may pose a significant barrier to successful gene therapy to the lung. In this work, we evaluated the diffusion of nanospheres and cationic DOTAP lipoplexes through CF sputum and the influence of CF mucus components on the physicochemical properties and gene expression of cationic DOTAP lipoplexes and neutral, pegylated GL67 lipoplexes. The number of particles transported through the sputum was extremely low (<0.5%) and strongly depending on the size of the particles, with almost no transport for the largest nanospheres (560 nm). For small particles (<150 nm), the low transport was primarily due to the long distance they have to travel through the sputum, while for larger particles also sterical obstruction was responsible for the low transport. Upon exposure of the cationic DOTAP lipoplexes to albumin, linear DNA or mucin (at concentration ratios expected to occur in vivo) a significant decrease in gene transfection activity was observed. This was primarily due to aggregation of the lipoplexes. However, exposure of pegylated GL67 lipoplexes to the same components did not affect their gene transfection activity. Indeed, it was determined that CF mucus components did not interact significantly with these neutral, pegylated GL67 lipoplexes.