Protein kinase C-α and arginase I mediate pneumolysin-induced pulmonary endothelial hyperpermeability

Rudolf Lucas, Guang Yang, Boris A Gorshkov, Evgeny Alexandrovich Zemskov, Supriya Sridhar, Umapathy N Siddaramappa, Agnieszka Jezierska-Drutel, Irina B. Alieva, Martin Leustik, Hamid Hossain, Bernhard Fischer, John D. Catravas, Alexander Dmitriyevich Verin, Jean François Pittet, Ruth B Caldwell, Timothy J. Mitchell, Stephen D. Cederbaum, David J Fulton, Michael A. Matthay, Robert William CaldwellMaritza Josefina Romero Lucas, Trinad Chakraborty

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Antibiotics-induced release of the pore-forming virulence factor pneumolysin (PLY) in patients with pneumococcal pneumonia results in its presence days after lungs are sterile and is a major factor responsible for the induction of permeability edema. Here we sought to identify major mechanisms mediating PLY-induced endothelial dysfunction. We evaluated PLY-induced endothelial hyperpermeability in human lung microvascular endothelial cells (HL-MVECs) and human lung pulmonary artery endothelial cells in vitro and in mice instilled intratracheally with PLY. PLY increases permeability in endothelial monolayers by reducing stable and dynamic microtubule content and modulating VE-cadherin expression. These events, dependent upon an increased calcium influx, are preceded by protein kinase C (PKC)-α activation, perturbation of the RhoA/Rac1 balance, and an increase in myosin light chain phosphorylation. At later time points, PLY treatment increases the expression and activity of arginase in HL-MVECs. Arginase inhibition abrogates and suppresses PLY-induced endothelial barrier dysfunction by restoring NO generation. Consequently, a specific PKC-α inhibitor and the TNF-derived tonoplast intrinsic protein peptide, which blunts PLY-induced PKC-α activation, are able to prevent activation of arginase in HL-MVECs and to reduce PLY-induced endothelial hyperpermeability in mice. Arginase I (AI) +/-/arginase II (AII)-/- C57BL/6 mice, displaying a significantly reduced arginase I expression in the lungs, are significantly less sensitive to PLY-induced capillary leak than their wild-type or AI +/+/AII-/- counterparts, indicating an important role for arginase I in PLY-induced endothelial hyperpermeability. These results identify PKC-α and arginase I as potential upstream and downstream therapeutic targets in PLY-induced pulmonary endothelial dysfunction.

Original languageEnglish (US)
Pages (from-to)445-453
Number of pages9
JournalAmerican journal of respiratory cell and molecular biology
Volume47
Issue number4
DOIs
StatePublished - Oct 1 2012

Fingerprint

Arginase
Protein Kinase C
Lung
Endothelial cells
Endothelial Cells
Chemical activation
Streptococcus pneumoniae plY protein
Permeability
Pneumococcal Pneumonia
Myosin Light Chains
Phosphorylation
Protein C Inhibitor
Virulence Factors
Protein Kinase Inhibitors
Inbred C57BL Mouse
Microtubules
Pulmonary Artery
Monolayers
Edema

Keywords

  • Arginase
  • PKC
  • Pneumococcus
  • Pneumolysin
  • TNF

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

Cite this

Protein kinase C-α and arginase I mediate pneumolysin-induced pulmonary endothelial hyperpermeability. / Lucas, Rudolf; Yang, Guang; Gorshkov, Boris A; Zemskov, Evgeny Alexandrovich; Sridhar, Supriya; Siddaramappa, Umapathy N; Jezierska-Drutel, Agnieszka; Alieva, Irina B.; Leustik, Martin; Hossain, Hamid; Fischer, Bernhard; Catravas, John D.; Verin, Alexander Dmitriyevich; Pittet, Jean François; Caldwell, Ruth B; Mitchell, Timothy J.; Cederbaum, Stephen D.; Fulton, David J; Matthay, Michael A.; Caldwell, Robert William; Romero Lucas, Maritza Josefina; Chakraborty, Trinad.

In: American journal of respiratory cell and molecular biology, Vol. 47, No. 4, 01.10.2012, p. 445-453.

Research output: Contribution to journalArticle

Lucas, R, Yang, G, Gorshkov, BA, Zemskov, EA, Sridhar, S, Siddaramappa, UN, Jezierska-Drutel, A, Alieva, IB, Leustik, M, Hossain, H, Fischer, B, Catravas, JD, Verin, AD, Pittet, JF, Caldwell, RB, Mitchell, TJ, Cederbaum, SD, Fulton, DJ, Matthay, MA, Caldwell, RW, Romero Lucas, MJ & Chakraborty, T 2012, 'Protein kinase C-α and arginase I mediate pneumolysin-induced pulmonary endothelial hyperpermeability', American journal of respiratory cell and molecular biology, vol. 47, no. 4, pp. 445-453. https://doi.org/10.1165/rcmb.2011-0332OC
Lucas, Rudolf ; Yang, Guang ; Gorshkov, Boris A ; Zemskov, Evgeny Alexandrovich ; Sridhar, Supriya ; Siddaramappa, Umapathy N ; Jezierska-Drutel, Agnieszka ; Alieva, Irina B. ; Leustik, Martin ; Hossain, Hamid ; Fischer, Bernhard ; Catravas, John D. ; Verin, Alexander Dmitriyevich ; Pittet, Jean François ; Caldwell, Ruth B ; Mitchell, Timothy J. ; Cederbaum, Stephen D. ; Fulton, David J ; Matthay, Michael A. ; Caldwell, Robert William ; Romero Lucas, Maritza Josefina ; Chakraborty, Trinad. / Protein kinase C-α and arginase I mediate pneumolysin-induced pulmonary endothelial hyperpermeability. In: American journal of respiratory cell and molecular biology. 2012 ; Vol. 47, No. 4. pp. 445-453.
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AU - Zemskov, Evgeny Alexandrovich

AU - Sridhar, Supriya

AU - Siddaramappa, Umapathy N

AU - Jezierska-Drutel, Agnieszka

AU - Alieva, Irina B.

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AU - Caldwell, Ruth B

AU - Mitchell, Timothy J.

AU - Cederbaum, Stephen D.

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