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Recombinant Tissue Plasminogen Activator (r-tPA) Induces In-Vitro Human Neutrophil Migration via Low Density Lipoprotein Receptor-Related Protein 1 (LRP-1)

Thrombolysis is the gold standard treatment for acute ischemic stroke. Besides its fibrinolytic role, recombinant tissue plasminogen activator (r-tPA) holds several non-fibrinolytic functions. Here, we investigated the potential role of r-tPA on human primary neutrophil migration in vitro. By means...

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Autores principales: Liberale, Luca, Bertolotto, Maria, Minetti, Silvia, Contini, Paola, Verzola, Daniela, Ameri, Pietro, Ghigliotti, Giorgio, Pende, Aldo, Camici, Giovanni G., Carbone, Federico, Montecucco, Fabrizio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7582901/
https://www.ncbi.nlm.nih.gov/pubmed/32977685
http://dx.doi.org/10.3390/ijms21197014
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author Liberale, Luca
Bertolotto, Maria
Minetti, Silvia
Contini, Paola
Verzola, Daniela
Ameri, Pietro
Ghigliotti, Giorgio
Pende, Aldo
Camici, Giovanni G.
Carbone, Federico
Montecucco, Fabrizio
author_facet Liberale, Luca
Bertolotto, Maria
Minetti, Silvia
Contini, Paola
Verzola, Daniela
Ameri, Pietro
Ghigliotti, Giorgio
Pende, Aldo
Camici, Giovanni G.
Carbone, Federico
Montecucco, Fabrizio
author_sort Liberale, Luca
collection PubMed
description Thrombolysis is the gold standard treatment for acute ischemic stroke. Besides its fibrinolytic role, recombinant tissue plasminogen activator (r-tPA) holds several non-fibrinolytic functions. Here, we investigated the potential role of r-tPA on human primary neutrophil migration in vitro. By means of modified Boyden chamber migration assay and checkerboard analysis we showed a dose-dependent chemotactic effect of r-TPA with a maximum effect reached by 0.03 mg/mL (0.003–1 mg/mL). Pre-incubation with MAP kinases inhibitors allowed the identification of PI3K/Akt, but not ERK1/2 as the intracellular pathway mediating the observed effects. Furthermore, by means of real-time PCR, immunocytochemistry and cytofluorimetry we demonstrated that the r-tPA receptor low density lipoprotein receptor-related protein 1 (LRP-1) is synthetized and expressed by neutrophils in response to r-tPA and TNF-α. Inhibition of LRP-1 by receptor-associated protein (RAP), prevented r-tPA-mediated F-actin polymerization, migration and signal through Akt but not ERK1/2. Lastly, also neutrophil degranulation in response to r-tPA seems to be mediated by LRP-1 under adhesion conditions. In conclusion, we show that r-tPA induces neutrophil chemotaxis through LRP-1/Akt pathway. Blunting r-tPA-mediated neutrophil activation might be beneficial as an adjuvant therapy to thrombolysis in this setting.
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spelling pubmed-75829012020-10-28 Recombinant Tissue Plasminogen Activator (r-tPA) Induces In-Vitro Human Neutrophil Migration via Low Density Lipoprotein Receptor-Related Protein 1 (LRP-1) Liberale, Luca Bertolotto, Maria Minetti, Silvia Contini, Paola Verzola, Daniela Ameri, Pietro Ghigliotti, Giorgio Pende, Aldo Camici, Giovanni G. Carbone, Federico Montecucco, Fabrizio Int J Mol Sci Article Thrombolysis is the gold standard treatment for acute ischemic stroke. Besides its fibrinolytic role, recombinant tissue plasminogen activator (r-tPA) holds several non-fibrinolytic functions. Here, we investigated the potential role of r-tPA on human primary neutrophil migration in vitro. By means of modified Boyden chamber migration assay and checkerboard analysis we showed a dose-dependent chemotactic effect of r-TPA with a maximum effect reached by 0.03 mg/mL (0.003–1 mg/mL). Pre-incubation with MAP kinases inhibitors allowed the identification of PI3K/Akt, but not ERK1/2 as the intracellular pathway mediating the observed effects. Furthermore, by means of real-time PCR, immunocytochemistry and cytofluorimetry we demonstrated that the r-tPA receptor low density lipoprotein receptor-related protein 1 (LRP-1) is synthetized and expressed by neutrophils in response to r-tPA and TNF-α. Inhibition of LRP-1 by receptor-associated protein (RAP), prevented r-tPA-mediated F-actin polymerization, migration and signal through Akt but not ERK1/2. Lastly, also neutrophil degranulation in response to r-tPA seems to be mediated by LRP-1 under adhesion conditions. In conclusion, we show that r-tPA induces neutrophil chemotaxis through LRP-1/Akt pathway. Blunting r-tPA-mediated neutrophil activation might be beneficial as an adjuvant therapy to thrombolysis in this setting. MDPI 2020-09-23 /pmc/articles/PMC7582901/ /pubmed/32977685 http://dx.doi.org/10.3390/ijms21197014 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Liberale, Luca
Bertolotto, Maria
Minetti, Silvia
Contini, Paola
Verzola, Daniela
Ameri, Pietro
Ghigliotti, Giorgio
Pende, Aldo
Camici, Giovanni G.
Carbone, Federico
Montecucco, Fabrizio
Recombinant Tissue Plasminogen Activator (r-tPA) Induces In-Vitro Human Neutrophil Migration via Low Density Lipoprotein Receptor-Related Protein 1 (LRP-1)
title Recombinant Tissue Plasminogen Activator (r-tPA) Induces In-Vitro Human Neutrophil Migration via Low Density Lipoprotein Receptor-Related Protein 1 (LRP-1)
title_full Recombinant Tissue Plasminogen Activator (r-tPA) Induces In-Vitro Human Neutrophil Migration via Low Density Lipoprotein Receptor-Related Protein 1 (LRP-1)
title_fullStr Recombinant Tissue Plasminogen Activator (r-tPA) Induces In-Vitro Human Neutrophil Migration via Low Density Lipoprotein Receptor-Related Protein 1 (LRP-1)
title_full_unstemmed Recombinant Tissue Plasminogen Activator (r-tPA) Induces In-Vitro Human Neutrophil Migration via Low Density Lipoprotein Receptor-Related Protein 1 (LRP-1)
title_short Recombinant Tissue Plasminogen Activator (r-tPA) Induces In-Vitro Human Neutrophil Migration via Low Density Lipoprotein Receptor-Related Protein 1 (LRP-1)
title_sort recombinant tissue plasminogen activator (r-tpa) induces in-vitro human neutrophil migration via low density lipoprotein receptor-related protein 1 (lrp-1)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7582901/
https://www.ncbi.nlm.nih.gov/pubmed/32977685
http://dx.doi.org/10.3390/ijms21197014
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