Ultrasound-guided transfection of claudin-5 improves lung endothelial barrier function in lung injury without impairing innate immunity

Abstract

In acute lung injury, the lung endothelial barrier is compromised. Loss of endothelial barrier integrity occurs in association with decreased levels of the tight junction protein claudin-5. Restoration of their levels by gene transfection may improve the vascular barrier but how to limit transfection solely to regions of the lung that are injured is unknown. We hypothesized that thoracic ultrasound in combination with intravenous microbubbles (USMB) could be used to achieve regional gene transfection in injured lung regions and improve endothelial barrier function. Since air blocks ultrasound energy, insonation of the lung is only achieved at areas of lung injury (edema, atelectasis); healthy lung is spared. Cavitation of the microbubbles achieves local tissue transfection. Here we demonstrate successful ultrasound-microbubble (USMB)-mediated gene transfection in the injured lungs of mice. After thoracic insonation, transfection was confined to the lung and only occurred in the setting of injured (but not healthy) lung. In a mouse model of acute lung injury, we observed down-regulation of endogenous claudin-5 and an acute improvement in lung vascular leakage and in oxygenation after claudin-5-over-expression by transfection. The improvement occurred without any impairment of the immune response as measured by pathogen clearance, alveolar cytokines and lung histology. In conclusion, USMB-mediated transfection targets injured lung regions and is a novel approach in the treatment of lung injury.

Secretome profiling of human epithelial cells exposed to cigarette smoke extract and their effect on human lung microvascular endothelial cells

“…Caveolin-1, a transmembrane protein, and claudin-5, a tight junction protein, both of which are abundant in lung endothelial cells, play important roles in regulating vascular permeability 29 , 30 . Cav1 −/− mutation can increase extravascular lung water by limiting water transport, thereby leading to pulmonary edema 31 , while overexpression of claudin-5 reverses oxygenation and modulates vascular leakage in a rat model of acute respiratory distress syndrome 32 . Therefore, the CS-exposed secretome of lung epithelial cells may increase vascular permeability.…”