Endothelial Knockdown of the Tumor Suppressor, , Increases Inflammation in Ventilator-induced Lung Injury

Overview

Journal Am J Physiol Lung Cell Mol Physiol

Publisher American Physiological Society

Specialties Cell Biology
Molecular Biology
Physiology
Pulmonary Medicine

Date 2024 Apr 2

PMID 38563965

Authors

Zhenguo Zeng

Eltyeb Abdelwahid

Weiguo Chen

Christian Ascoli

Trinh Pham

Jeffrey R Jacobson

Steven M Dudek

Viswanathan Natarajan

C Marcelo Aldaz

Roberto F Machado

Sunit Singla

Affiliations

Soon will be listed here.

Abstract

Chronic cigarette smoke exposure decreases lung expression of which is known to protect the endothelial barrier during infectious models of acute respiratory distress syndrome (ARDS). Proteomic analysis of -silenced endothelial cells (ECs) was done using tandem mass tag mass spectrometry (TMT-MS). -silenced ECs as well as those isolated from endothelial cell knockout (EC KO) mice were subjected to cyclic stretch (18% elongation, 0.5 Hz, 4 h). Cellular lysates and media supernatant were harvested for assays of cellular signaling, protein expression, and cytokine release. These were repeated with dual silencing of and zyxin. Control and EC KO mice were subjected to high tidal volume ventilation. Bronchoalveolar lavage fluid and mouse lung tissue were harvested for cellular signaling, cytokine secretion, and histological assays. TMT-MS revealed upregulation of zyxin expression during knockdown which predicted a heightened inflammatory response to mechanical stretch. -silenced ECs and ECs isolated from EC mice displayed significantly increased cyclic stretch-mediated secretion of various cytokines (IL-6, KC/IL-8, IL-1β, and MCP-1) relative to controls. This was associated with increased ERK and JNK phosphorylation but decreased p38 mitogen-activated kinases (MAPK) phosphorylation. EC KO mice subjected to VILI sustained a greater degree of injury than corresponding controls. Silencing of zyxin during knockdown abrogated stretch-induced increases in IL-8 secretion but not in IL-6. Loss of function in ECs is associated with a heightened inflammatory response during mechanical stretch that is associated with increased MAPK phosphorylation and appears, in part, to be dependent on the upregulation of zyxin. Prior tobacco smoke exposure is associated with an increased risk of acute respiratory distress syndrome (ARDS) during critical illness. Our laboratory is investigating one of the gene expression changes that occurs in the lung following smoke exposure: downregulation. Here we describe changes in protein expression associated with knockdown and its influence on ventilator-induced ARDS in a mouse model.

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