From Texas Lung Injury Institute: Intrapleural adenoviral delivery of human PAI-1 exacerbates TCN-induced pleural injury in rabbits

http://ajrcmb.atsjournals.org/content/early/2012/09/19/rcmb.2012-0183OC.full.pdf+html


Intrapleural adenoviral delivery of human PAI-1 exacerbates TCN-induced pleural injury in rabbits 

Sophia Karandashova, Galina Florova, Ali O. Azghani, Andrey A. Komissarov, Kathy
Koenig, Torry A. Tucker, Timothy C. Allen, Kris Stewart, Amy Tvinnereim, and Steven
Idell

The Texas Lung Injury Institute of The University of Texas Health Science Center at Tyler,
11937 US Highway 271, Tyler, TX 75708-3154, USA;
The University of Texas at Tyler, Tyler TX.




ABSTRACT: Elevated levels of plasminogen activator inhibitor 1 (PAI-1) are associated with pleural

injury but its effects on pleural organization remain unclear. A method of adenovirus-mediated

delivery of genes of interest, expressed under a cytomegalovirus promoter, to the rabbit pleura

was developed and used with lacZ and human (h) PAI-1. Histology, β-galactosidase staining,

Western blot, enzymatic and immunohistochemical analyses of pleural fluids (PFs), lavages, and

pleural mesothelial cells were used to evaluate the efficiency and effects of transduction.

Transduction was selective and limited to the pleural mesothelial monolayer. Intrapleural

expression of both genes was transient, with peak expression at 4-5 days. At day 5, hPAI-1 (40-

80 and 200-400 nM of active and total hPAI-1 in lavages, respectively) caused no overt pleural

injury, effusions or fibrosis. Adenovirus-mediated delivery of hPAI-1 with subsequent

tetracycline-induced pleural injury resulted in a significant exacerbation of the pleural fibrosis

observed at day 5 (P=0.029 and 0.021 versus vehicle and adenoviral controls, respectively).

Intrapleural fibrinolytic therapy (IPFT) with plasminogen activators was effective in both

animals overexpressing hPAI-1 and controls with tetracycline-injury alone. An increase in

intrapleural active PAI-1 (from 10-15 in controls to 20-40 nM in hPAI-1 overexpressing

animals) resulted in increased formation of PAI-1/plasminogen activator complexes in vivo. The

decrease in intrapleural plasminogen-activating activity observed at 10-40 min after IPFT

correlates linearly with the initial level of active PAI-1. Therefore, active PAI-1 in PFs affects

the outcome of IPFT, and may be both a biomarker of pleural injury and a molecular target for

its treatment.