Fibrin deposition that results in pleural loculation, scarring, or lung injury is difficult to treat and is associated with a high level of morbidity and mortality. Fibrinolytic therapy activates the endogenous fibrinolytic system, resolving adhesions and complex fibrinous deposits that sequester pockets of inflammation (loculations), thus improving fluid drainage and clinical outcomes. We have identified several major elements of the mechanism of intrapleural fibrinolysis, confirmed and validated active plasminogen activator inhibitor 1 (PAI-1) as a biomarker and molecular target for therapeutic intervention. We are developing novel, safe PAI-1 targeting fibrinolytic therapies, and innovative diagnostic assays to provide better, safer treatments for patients with pleural loculation and lung injury.
- Regulation of innate immunity and inflammation
- Neutrophils, macrophages, endothelial, and epithelial cells - activation and apoptosis
- Pro-inflammatory receptors - regulation and signaling
- Fibrinolytic therapy in infectious animal models of pleural injury and fibrosis, as well as in empyema in humans
- Molecular biomarkers and therapeutic targets in pleural fibrosis to increase the efficacy of fibrinolytic therapy
- Using modern, non-invasive imaging approaches and developing new companion assays to evaluate the severity of pleural injury and monitor fibrinolytic potential
- Molecular mechanisms of fibrinolysis in empyema and inhalation smoke-induced acute lung injury (ISALI)
Approximately 80,000 patients in the US and UK develop complicated parapneumonic pleural effusions (CPE) and empyema (EMP) each year. Both are common clinical problems associated with a mortality of about 20%, serious morbidity, and annual costs of $500M. The incidence of pleural infections is 8-fold greater than that of cystic fibrosis, 5-fold than that of idiopathic pulmonary fibrosis; patient mortality exceeds that of myocardial infarction or community acquired pneumonia. While surgery can be lifesaving, it is invasive, of unproven efficacy, and many patients are not operative candidates. A less invasive alternative, intrapleural fibrinolytic therapy (IPFT), which is pharmacologic drainage of the pleural space, remains non-standardized due to our lack of understanding of the mechanisms governing fibrinolysis. We have identified active plasminogen activator inhibitor 1 (PAI-1) as a biomarker for the outcome of pleural injury, validated active PAI-1 as a therapeutic target, and identified 3 novel strategies that resulted in an 8-fold decrease in the dose necessary for effective fibrinolysis. Employing non-invasive computed tomography scanning and ultrasonography allowed us to determine the time needed for effective intrapleural fibrinolysis. These results have built a solid mechanistic foundation for expanding our understanding of the mechanisms of fibrinolysis to infectious models of pleural injury and other forms of acute and chronic fibrosis such as ISALI, and sepsis. Our research results in novel interventions, as well as imaging and diagnostic approaches that are ready to enter the clinic for patients with pleural loculation and lung injury, who are difficult to treat, prone to poor outcomes, and for whom safe and effective therapy is not currently available.
Marudamuthu AS, Shetty SK, Bhandary YP, Karandashova S, Thompson M, Sathish V, Florova G, Hogan TB, Pabelick CM, Prakash YS, Tsukasaki Y, Fu J, Ikebe M, Idell S, Shetty S. Plasminogen activator inhibitor-1 suppresses profibrotic responses in fibroblasts from fibrotic lungs. J Biol Chem. 2015 Apr 10;290(15):9428-41. PMID:256488922.
Florova G, Azghani A, Karandashova S, Schaefer C, Koenig K, Stewart-Evans K, Declerck PJ, Idell S, Komissarov AA. Targeting of plasminogen activator inhibitor 1 improves fibrinolytic therapy for tetracycline-induced pleural injury in rabbits. Am J Respir Cell Mol Biol. 2015 Apr;52(4):429-37 PMID:25140386.
Salem SM, Kancharla P, Florova G, Gupta S, Lu W, Reynolds KA. Elucidation of final steps of the marineosins biosynthetic pathway through identification and characterization of the corresponding gene cluster. J Am Chem Soc. 2014 Mar 26;136(12):4565-74. PMID: 24575817
Komissarov AA, Florova G, Azghani A, Karandashova S, Kurdowska AK, Idell S. Active á-macroglobulin is a reservoir for urokinase after fibrinolytic therapy in rabbits with tetracycline-induced pleural injury and in human pleural fluids. Am J Physiol Lung Cell Mol Physiol. 2013 Nov 15;305(10):L682-92. PMID: 2399717
Florova G, Karandashova S, Declerck PJ, Idell S, Komissarov AA. Remarkable stabilization of plasminogen activator inhibitor 1 in a "molecular sandwich" complex. Biochemistry. 2013 Jul 9;52(27):4697-709.. Epub 2013 Jun 25. PMID:23734661
Bonnett SA, Whicher JR, Papireddy K, Florova G, Smith JL, Reynolds KA. Structural and stereochemical analysis of a modular polyketide synthase ketoreductase domain required for the generation of a cis-alkene. Chem Biol. 2013 Jun 20;20(6):772-83. PMID:23790488
Karandashova S, Florova G, Azghani AO, Komissarov AA, Koenig K, Tucker TA, Allen TC, Stewart K, Tvinnereim A, Idell S. Intrapleural adenoviral delivery of human plasminogen activator inhibitor-1 exacerbates tetracycline-induced pleural injury in rabbits. Am J Respir Cell Mol Biol. 2013 Jan;48(1):44-52. PMID:23002099
Komissarov AA, Stankowska D, Krupa A, Fudala R, Florova G, Florence J, Fol M, Allen TC, Idell S, Matthay MA, Kurdowska AK. Novel aspects of urokinase function in the injured lung: role of á2-macroglobulin. Am J Physiol Lung Cell Mol Physiol. 2012 Dec 15;303(12):L1037-45. PMID:23064953
Singh R, Mo S, Florova G, Reynolds KA. Streptomyces coelicolor RedP and FabH enzymes, initiating undecylprodiginine and fatty acid biosynthesis, exhibit distinct acyl-CoA and malonyl-acyl carrier protein substrate specificities. FEMS Microbiol Lett. 2012 Mar;328(1):32-8. doi: 10.1111/j.1574-6968.2011.02474.x. Epub 2012 Jan 4.
Komissarov AA, Florova G, Idell S. Effects of extracellular DNA on plasminogen activation and fibrinolysis. J Biol Chem. 2011 Dec 9;286(49):41949-62. PMID:21976662