Adenoviral gene transfer of a mutant surfactant enzyme ameliorates pseudomonas-induced lung injury

Surfactant deficiency is an important contributor to the acute respiratory distress syndrome, a disorder that commonly occurs after bacterial sepsis. CTP: phosphocholine cytidylyltransferase (CCTa) is the rate-limiting enzyme required for the biosynthesis of dipalmitoylphosphatidylcholine (DPPC), the major phospholipid of surfactant. In this study, a cDNA encoding a novel, calpain-resistant mutant CCTa enzyme was delivered intratracheally in mice using a replication-deficient adenovirus 5 CTP: phosphocholine cytidylyltransferase construct (Ad5-CCTPenta) in models of bacterial sepsis. Ad5-CCTPenta gene transfer produced high-level CCTa gene expression, increased alveolar surfactant (DPPC) levels and improved lung surface tension and pressure-volume relationships relative to control mice. Pseudomonas aeruginosa (PA103) decreased DPPC synthesis, in part, via calpain-mediated degradation of CCTa. Deleterious effects of Pseudomonas on surfactant were lessened after infection with a mutant strain lacking the type III exotoxin, Exo U. Replication-deficient adenovirus 5 CTP: phosphocholine cytidylyltransferase gene delivery improved lung biophysical properties by optimizing surface activity in this Pseudomonas model of proteinase-mediated lung injury. The studies are the first demonstration of in vivo gene transfer of a lipogenic enzyme resulting in improved lung mechanics. The studies suggest that augmentation of DPPC synthesis via gene delivery of CCTa can attenuate impaired lung function in surfactant-deficient states such as bacterial sepsis.