Role of cGMP in Ventilator-Induced Lung Endothelial Barrier Dysfunction

Summary

Principal Investigator: David B Pearse
Abstract: DESCRIPTION (provided by applicant): Failure of the lung endothelial barrier leads to pulmonary edema and acute lung injury (ALI). Mechanical ventilation contributes to ALI mortality through excessive ventilatory stretch (VS), causing ventilator-induced lung injury (VILI). In animal models of VILI, increased endothelial nitric oxide (NO) production contributes to pulmonary endothelial barrier dysfunction. NO activates endothelial soluble guanylyl cyclase (sGC), producing cGMP but the effect of endothelial cGMP on VS-induced endothelial barrier function is unknown. Our preliminary experiments in a mouse model of VILI suggest that VS activates endothelial sGC to increase cGMP. After VS onset, sGC inhibition or stimulation attenuates or exacerbates barrier dysfunction, respectively, suggesting an injurious role for cGMP. In contrast, sGC activation before VS is barrier protective. Injurious VS also increases cGMP-activated phosphodiesterase 2A (PDE2A) mRNA and protein expression. PDE2A inhibition is protective. Inhaled bacterial lipopolysaccharide (LPS) also increases PDE2A expression and causes injury that is attenuated by either sGC or PDE2A inhibition. The overall goal of this application is to determine the effect of sGC-mediated cGMP production on endothelial permeability in VILI and in the combination of VILI with LPS inhalation to model septic ALI. We hypothesize that injurious VS increases endothelial 1) cGMP from NO-stimulated sGC and 2) PDE2A expression through activation of p38 and MAP kinase-activated protein kinase 2 (MK2) resulting in accelerated hydrolysis of endothelial barrier protective cAMP. In Aim 1, we plan to determine the relationships between VS, sGC activation, cytokine production, MAPK activation, cAMP and changes in pulmonary endothelial permeability in isolated mouse lungs, anesthetized ventilated mice, and monolayers of mouse lung microvascular endothelial cells subjected to injurious cyclic stretch, LPS and/or cytokine exposure. These experiments will utilize the novel sGC stimulator BAY 41-2272 and sGCa1-/- mice to manipulate lung sGC activity. In Aim 2, we will determine the roles of PDE2A and 3A in the complex effects of cGMP on pulmonary endothelial barrier function in lungs and endothelial monolayers subjected to injurious stretch LPS or cytokines. PDE mRNA, protein expression and activity will be studied. Specific inhibitors, a PDE2A shRNAmiR adenovirus to decrease endothelial PDE2A expression and PDE3A-/- mice will be employed in these experiments. In Aim 3, we plan to determine the upstream pathways responsible for the increased expression of PDE2A in VILI. Specifically, we plan to examine the roles of TNFa, p38, MK2 and NF?B using specific inhibitors, siRNA, as well as TNFa -/-, TNFa Receptor1-/-, MK2-/-, and p50-/- mice to dissect the involvement of each signaling pathway. These experiments will explore a novel mechanism of lung endothelial barrier dysfunction mediated by the simultaneous stimulation of sGC/cGMP and PDE2A expression by injurious levels of VS. PUBLIC HEALTH RELEVANCE: The goal of this grant proposal is to understand the cellular and biochemical mechanisms that cause the blood vessels in the lung to leak fluid when they are overly stretched by artificial ventilation. It has become increasingly clear that the breathing support we provide to critically ill patients helps to maintain life but at the same time can result in a lung injury that may eventually cause severe complications. The cells lining the blood vessels of the lung respond to mechanical distension in complex ways that could be modified by medications if the cellular effects are unraveled.
Funding Period: 2001-04-01 - 2013-12-31
more information: NIH RePORT

Top Publications

  1. ncbi Role of protein kinase G in barrier-protective effects of cGMP in human pulmonary artery endothelial cells
    Aigul Moldobaeva
    Division of Pulmonary and Critical Care Medicine, Department of Medicine, Hopkins Bayview Medical Center, 5501 Hopkins Bayview Circle, Baltimore, MD 21224, USA
    Am J Physiol Lung Cell Mol Physiol 290:L919-30. 2006
  2. ncbi The role of natriuretic peptide receptor-A signaling in unilateral lung ischemia-reperfusion injury in the intact mouse
    Jeffrey M Dodd-o
    Department of Anesthesia and Critical Care, School of Medicine, The Johns Hopkins Medical Institutions, Baltimore, MD 21287 9106, USA
    Am J Physiol Lung Cell Mol Physiol 294:L714-23. 2008
  3. ncbi Role of vasodilator-stimulated phosphoprotein in cGMP-mediated protection of human pulmonary artery endothelial barrier function
    Otgonchimeg Rentsendorj
    Department of Medicine, Division of Pulmonary and Critical Care Medicine, The Johns Hopkins University, Baltimore, MD 21224, USA
    Am J Physiol Lung Cell Mol Physiol 294:L686-97. 2008
  4. pmc Soluble guanylyl cyclase contributes to ventilator-induced lung injury in mice
    Eric P Schmidt
    Division of Pulmonary and Critical Care Medicine, Department of Medicine, The Johns Hopkins Medical Institutions, Baltimore, Maryland 21224, USA
    Am J Physiol Lung Cell Mol Physiol 295:L1056-65. 2008
  5. pmc Knockdown of lung phosphodiesterase 2A attenuates alveolar inflammation and protein leak in a two-hit mouse model of acute lung injury
    Otgonchimeg Rentsendorj
    Division of Pulmonary and Critical Care Medicine, Department of Medicine, Johns Hopkins Medical Institutions, Baltimore, MD 21224, USA
    Am J Physiol Lung Cell Mol Physiol 301:L161-70. 2011
  6. pmc Protein kinase G increases antioxidant function in lung microvascular endothelial cells by inhibiting the c-Abl tyrosine kinase
    R Scott Stephens
    Division of Pulmonary and Critical Care Medicine, Department of Medicine, Johns Hopkins University, Baltimore, Maryland, and
    Am J Physiol Cell Physiol 306:C559-69. 2014

Research Grants

  1. RAGE and Mechanisms of Vascular Dysfunction
    Shi Fang Yan; Fiscal Year: 2013

Detail Information

Publications6

  1. ncbi Role of protein kinase G in barrier-protective effects of cGMP in human pulmonary artery endothelial cells
    Aigul Moldobaeva
    Division of Pulmonary and Critical Care Medicine, Department of Medicine, Hopkins Bayview Medical Center, 5501 Hopkins Bayview Circle, Baltimore, MD 21224, USA
    Am J Physiol Lung Cell Mol Physiol 290:L919-30. 2006
    ....
  2. ncbi The role of natriuretic peptide receptor-A signaling in unilateral lung ischemia-reperfusion injury in the intact mouse
    Jeffrey M Dodd-o
    Department of Anesthesia and Critical Care, School of Medicine, The Johns Hopkins Medical Institutions, Baltimore, MD 21287 9106, USA
    Am J Physiol Lung Cell Mol Physiol 294:L714-23. 2008
    ..These data suggest that lung IR triggered ANP release and altered endothelial signaling so that NPR-A activation caused increased pulmonary endothelial permeability...
  3. ncbi Role of vasodilator-stimulated phosphoprotein in cGMP-mediated protection of human pulmonary artery endothelial barrier function
    Otgonchimeg Rentsendorj
    Department of Medicine, Division of Pulmonary and Critical Care Medicine, The Johns Hopkins University, Baltimore, MD 21224, USA
    Am J Physiol Lung Cell Mol Physiol 294:L686-97. 2008
    ..Our results suggest that VASP phosphorylation does not explain the protective effect of cGMP/PKG(I) on H(2)O(2)-induced endothelial barrier dysfunction in HPAEC...
  4. pmc Soluble guanylyl cyclase contributes to ventilator-induced lung injury in mice
    Eric P Schmidt
    Division of Pulmonary and Critical Care Medicine, Department of Medicine, The Johns Hopkins Medical Institutions, Baltimore, Maryland 21224, USA
    Am J Physiol Lung Cell Mol Physiol 295:L1056-65. 2008
    ..We conclude that HV(T)-induced endothelial barrier dysfunction resulted from a simultaneous increase in NO/sGC-derived cGMP and PDE2A expression causing decreased cAMP...
  5. pmc Knockdown of lung phosphodiesterase 2A attenuates alveolar inflammation and protein leak in a two-hit mouse model of acute lung injury
    Otgonchimeg Rentsendorj
    Division of Pulmonary and Critical Care Medicine, Department of Medicine, Johns Hopkins Medical Institutions, Baltimore, MD 21224, USA
    Am J Physiol Lung Cell Mol Physiol 301:L161-70. 2011
    ..We conclude that in a mouse model of LPS/VILI, a synergistic increase in lung PDE2A expression increased lung iNOS and alveolar inflammation and contributed significantly to the ensuing acute lung injury...
  6. pmc Protein kinase G increases antioxidant function in lung microvascular endothelial cells by inhibiting the c-Abl tyrosine kinase
    R Scott Stephens
    Division of Pulmonary and Critical Care Medicine, Department of Medicine, Johns Hopkins University, Baltimore, Maryland, and
    Am J Physiol Cell Physiol 306:C559-69. 2014
    ..Inhibition of c-Abl by active PKGI may be the downstream mechanism underlying PKGI-mediated antioxidant signaling. Tyrosine kinase inhibitors may represent a novel therapeutic approach in oxidant-induced ALI. ..

Research Grants30

  1. RAGE and Mechanisms of Vascular Dysfunction
    Shi Fang Yan; Fiscal Year: 2013
    ..Using novel and state-of-the-art techniques, floxed mice and molecular approaches to gene regulation, we are well-positioned to lead the study of RAGE in the next cycle of this Program. ..