Effects of Extracellular DNA on Plasminogen Activation and Fibrinolysis

Komissarov, Andrey A.; Florova, Galina; Idell, Steven

doi:10.1074/jbc.m111.301218

Cited by 63 publications

(101 citation statements)

References 94 publications

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“…PFs of rabbits with TCN-induced pleural injury (n ¼ 5) and TCN-induced injury enhanced by overexpressing hPAI-1 intrapleurally (n ¼ 4) were treated with an effective dose of scuPA (open symbols) or sctPA (solid symbols) (0.5 and 0.145 mg/kg, respectively) (37). PAI-1 activity was measured in PF collected before IPFT, as described previously (33). PA activity was measured in PF collected 10 minutes (circles), 20 minutes (squares), and 40 minutes (triangles) after IPFT, as described elsewhere (33).…”

Section: Discussionmentioning

confidence: 99%

“…To test the effects of intrapleural active PAI-1 on the PA activity of fibrinolysins during IPFT, the aliquots withdrawn at 10, 20, and 40 minutes were tested for their ability to activate human Glu-plasminogen, as previously described (33). The PA activity in PF at 10, 20, and 40 minutes was plotted versus the initial intrapleural concentration of active PAI-1 ( Figure 6).…”

Section: The Effects Of Mesothelial Hpai-1 Expression On Intrapleuralmentioning

confidence: 99%

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Intrapleural Adenoviral Delivery of Human Plasminogen Activator Inhibitor–1 Exacerbates Tetracycline-Induced Pleural Injury in Rabbits

Karandashova

Florova

Azghani

et al. 2013

Am J Respir Cell Mol Biol

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106

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Elevated concentrations 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 rabbit pleura was developed and used with lacZ and human (h) PAI-1. Histology, b-galactosidase staining, Western blotting, 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. The intrapleural expression of both genes was transient, with their peak expression at 4 to 5 days. On 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. The adenovirus-mediated delivery of hPAI-1 with subsequent tetracycline-induced pleural injury resulted in a significant exacerbation of the pleural fibrosis observed on Day 5 (P ¼ 0.029 and P ¼ 0.021 versus vehicle and adenoviral control samples, respectively). Intrapleural fibrinolytic therapy (IPFT) with plasminogen activators was effective in both animals overexpressing hPAI-1 and control animals with tetracycline injury alone. An increase in intrapleural active PAI-1 (from 10-15 nM in control animals to 20-40 nM in hPAI-1-overexpressing animals) resulted in the increased formation of PAI-1/plasminogen activator complexes in vivo. The decrease in intrapleural plasminogen-activating activity observed at 10 to 40 minutes after IPFT correlates linearly with the initial concentration 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.Keywords: pleural injury; plasminogen activator inhibitor-1; intrapleural fibrinolytic therapyThe incidence of complicated pleural infection and empyema, a serious infection of the pleural space often associated with pneumonia, is increasing in the United States (1) and other countries, in both adult and pediatric populations (2-5). The exact cause of this increase is unknown, although the increased prevalence of antibiotic-resistant bacteria, changes in empyema management, and changes in causative bacterial agents have been implicated (1,3,6). Pleural infections, empyema, or complicated parapneumonic effusions develop in approximately 80,000 patients in the United States and the United Kingdom annually (7). In the United Kingdom, a 20% mortality rate was reported for patients with empyema, and 20% of patients require surgical intervention after developing a pleural infection (7). When pleural effusions occur in association with high-grade inflammation, they can organize with the development of loculation, where an effusion becomes trapped behind partly fused visceral and parietal pleura, with pleural thickening (8-10). Persistent pleural loculation and fibrosis increase morbidity and mortality an...

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Section: Discussionmentioning

confidence: 99%

Section: The Effects Of Mesothelial Hpai-1 Expression On Intrapleuralmentioning

confidence: 99%

Intrapleural Adenoviral Delivery of Human Plasminogen Activator Inhibitor–1 Exacerbates Tetracycline-Induced Pleural Injury in Rabbits

Karandashova

Florova

Azghani

et al. 2013

Am J Respir Cell Mol Biol

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106

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“…Effect of Anti-PAI-1 mAbs and the Dosage of scuPA on Intrapleural Fibrinolytic Activity at 0-80 min after Administration of IPFT To determine whether PAI-1-targeted IPFT with 0.0625 mg/kg scuPA affects intrapleural fibrinolytic activity, PFs were withdrawn at 0-40 minutes and at 24 hours after IPFT and tested using an FITC-fibrin plate assay as previously described (29). Notably, there was no significant (P .…”

Section: Data Analysis and Statisticsmentioning

confidence: 99%

“…Although the baseline (before IPFT) fibrinolytic activity was suppressed (close or equal to the limit of detection by FITC-fibrin assay), supplementation with , and 40 minutes after administration of 0.0625 mg/kg scuPA either with anti-PAI-1 mAbs (0.5 mg/kg; red triangles; n = 5) or mouse IgG (0.5 mg/kg; blue triangles; n = 6). Aliquots of PF withdrawn 10-40 minutes after intrapleural administration of the fibrinolysin were analyzed with FITC-fibrin film assay as described previously (29). (B) Aliquots of PF withdrawn before (zero point; 0 h) and at 24 hours after injection of the fibrinolysin were analyzed via FITC-fibrin film assay (29) with (uPA) or without (no uPA) addition of exogenous two-chain uPA (tcuPA; 5 nM).…”

Section: Data Analysis and Statisticsmentioning

confidence: 99%

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Targeting of Plasminogen Activator Inhibitor 1 Improves Fibrinolytic Therapy for Tetracycline-Induced Pleural Injury in Rabbits

Florova

Azghani

Karandashova

et al. 2015

Am J Respir Cell Mol Biol

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133

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Endogenous active plasminogen activator inhibitor 1 (PAI-1) was targeted in vivo with monoclonal antibodies (mAbs) that redirect its reaction with proteinases to the substrate branch. mAbs were used as an adjunct to prourokinase (single-chain [sc] urokinase [uPA]) intrapleural fibrinolytic therapy (IPFT) of tetracycline-induced pleural injury in rabbits. Outcomes of scuPA IPFT (0.25 or 0.0625 mg/kg) with 0.5 mg/kg of mouse IgG or mAbs (MA-33H1F7 and MA-8H9D4) were assessed at 24 hours. Pleural fluid (PF) was collected at 0, 10, 20, and 40 minutes and 24 hours after IPFT and analyzed for plasminogen activating (PA), uPA, fibrinolytic activities, levels of total plasmin/plasminogen, a-macroglobulin (aM), mAbs/IgG antigens, free active uPA, and aM/uPA complexes. Anti-PAI-1 mAbs, but not mouse IgG, delivered with an eightfold reduction in the minimal effective dose of scuPA (from 0.5 to 0.0625 mg/kg), improved the outcome of IPFT (P , 0.05). mAbs and IgG were detectable in PFs at 24 hours. Compared with identical doses of scuPA alone or with IgG, treatment with scuPA and anti-PAI-1 mAbs generated higher PF uPA amidolytic and PA activities, faster formation of aM/uPA complexes, and slower uPA inactivation. However, PAI-1 targeting did not significantly affect intrapleural fibrinolytic activity or levels of total plasmin/plasminogen and aM antigens. Targeting PAI-1 did not induce bleeding, and rendered otherwise ineffective doses of scuPA able to improve outcomes in tetracycline-induced pleural injury. PAI-1-neutralizing mAbs improved IPFT by increasing the durability of intrapleural PA activity. These results suggest a novel, well-tolerated IPFT strategy that is tractable for clinical development. Keywords: plasminogen activator inhibitor 1; fibrinolytic therapy; animal model; prourokinase; monoclonal antibodies Clinical RelevanceOrganizing pleural injury remains an important clinical problem for which fibrinolytic therapy has been used with variable results for children and adults. This study demonstrates, for the first time, that the targeting of active plasminogen activator inhibitor 1 enhances the ability of relatively low doses of intrapleural single-chain urokinase to clear pleural effusions after induction of organizing injury. This work defines a new, well-tolerated approach for intrapleural fibrinolytic therapy that is promising and tractable for clinical trial testing.The results of Multicenter Intrapleural Sepsis Trials 1 and 2 demonstrated that intrapleural fibrinolytic therapy (IPFT) with either streptokinase, or tissue-type plasminogen activator (tPA) alone were ineffective (1, 2). In contrast, there is a growing body of clinical reports demonstrating the successful use of IPFT, including tPA,. It is likely that the disparate results of IPFT trials, which are largely successful in children (2, 6) and variably effective in adults (6, 7), relate to the lack of formal toxicological and dose-escalation studies, resulting in empiric dosing. A further impediment to the field is an incomplete underst...

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Perfluorochemical‐facilitated plasminogen activator delivery to the airways: A novel treatment for inhalational smoke‐induced acute lung injury

Wolfson

Enkhbaatar

Fukuda

et al. 2020

Clinical & Translational Med

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Background Effective clinical management of airway clot and fibrinous cast formation of severe inhalational smoke‐induced acute lung injury (ISALI) is lacking. Aerosolized delivery of tissue plasminogen activator (tPA) is confounded by airway bleeding; single‐chain urokinase plasminogen activator (scuPA) moderated this adverse effect and supported transient improvement in gas exchange and lung mechanics. However, neither aerosolized plasminogen activator (PA) yielded durable improvements in physiologic responses or reduction in cast burden. Here, we hypothesized that perfluorochemical (PFC) liquids would facilitate PA distribution and sustain improvements in physiologic outcomes in ISALI. Methods Spontaneously breathing adult sheep (n = 36) received anesthesia and analgesia and were instrumented, exposed to cotton smoke inhalation, and supported by mechanical ventilation for 48 h. Groups (n = 6/group) were studied without supplemental treatment, or, starting 4 h post injury, they received intratracheal low volume (8 mL) PFC liquid alone or a dose range of tPA/PFC or scuPA/PFC suspensions (4 or 8 mg in 8 mL PFC) every 8 h. Outcomes were evaluated by sequential measurements of cardiopulmonary parameters, lung histomorphology, and biochemical analyses of bronchoalveolar lavage fluid. Results Dose‐response and PA‐type comparisons of outcomes demonstrated sustained superiority with low‐volume PFC suspensions of scuPA over tPA or PFC alone, favoring the highest dose of scuPA/PFC suspension over lower doses, without airway bleeding. Conclusions We propose that this improved profile over previously reported aerosolized delivery is likely related to improved dose distribution. Sustained salutary responses to scuPA/PFC suspension delivery in this translational model are encouraging and support the possibility that the observed outcomes could be of clinical importance.

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Effects of Extracellular DNA on Plasminogen Activation and Fibrinolysis

Cited by 63 publications

References 94 publications

Intrapleural Adenoviral Delivery of Human Plasminogen Activator Inhibitor–1 Exacerbates Tetracycline-Induced Pleural Injury in Rabbits

Intrapleural Adenoviral Delivery of Human Plasminogen Activator Inhibitor–1 Exacerbates Tetracycline-Induced Pleural Injury in Rabbits

Targeting of Plasminogen Activator Inhibitor 1 Improves Fibrinolytic Therapy for Tetracycline-Induced Pleural Injury in Rabbits

Perfluorochemical‐facilitated plasminogen activator delivery to the airways: A novel treatment for inhalational smoke‐induced acute lung injury

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