Cigarette smoke impairs clearance of apoptotic cells through oxidant-dependent activation of RhoA.
BACKGROUND
Cigarette smoke (CS) is the primary cause of chronic obstructive pulmonary disease (COPD), an effect that is, in part, due to intense oxidant stress. Clearance of apoptotic cells (efferocytosis) is a critical regulator of lung homeostasis, which is defective in smokers and in patients with COPD, suggesting a role in disease pathogenesis.


OBJECTIVE
We hypothesized that CS would impair efferocytosis through oxidant-dependent activation of RhoA, a known inhibitor of this process.


METHODS
We investigated the effect of CS on efferocytosis in vivo and ex vivo, using acute, subacute, and long-term mouse exposure models.


RESULTS
Acute and subacute CS exposure suppressed efferocytosis by alveolar macrophages in a dose-dependent, reversible, and cell type-independent manner, whereas more intense CS exposure had an irreversible effect. In contrast, CS did not alter ingestion through the Fc gamma receptor. The inhibitory effect of CS on apoptotic cell clearance depended on oxidants, because the effect was blunted in oxidant-resistant ICR mice, and was prevented by either genetic or pharmacologic antioxidant strategies in vivo and ex vivo. CS inhibited efferocytosis through oxidant-dependent activation of the RhoA-Rho kinase pathway because (1) CS activated RhoA, (2) antioxidants prevented RhoA activation by CS, and (3) inhibitors of the RhoA-Rho kinase pathway reversed the suppressive effect of CS on apoptotic cell clearance in vivo and ex vivo.


CONCLUSIONS
These findings advance the hypothesis that impaired efferocytosis may contribute to the pathogenesis of COPD and suggest the therapeutic potential of drugs targeting the RhoA-Rho kinase pathway.
Richens
Tiffany R
TR
University of Colorado Denver, Chronic Obstructive Pulmonary Disease Center, Division of Pulmonary Sciences and Critical Care Medicine, 12700 E. 19th Avenue, Aurora, CO 80045, USA.
Linderman
Derek J
DJ
Horstmann
Sarah A
SA
Lambert
Cherie
C
Xiao
Yi-Qun
YQ
Keith
Robert L
RL
Boé
Darren M
DM
Morimoto
Konosuke
K
Bowler
Russell P
RP
Day
Brian J
BJ
Janssen
William J
WJ
Henson
Peter M
PM
Vandivier
R William
RW
D000818:Animals; D017209:Apoptosis; D045744:Cell Line, Tumor; D006801:Humans; D016676:Macrophages, Alveolar / Q000187:drug effects* / Q000276:immunology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D018345:Mice, Knockout; D009504:Neutrophils; D018384:Oxidative Stress / Q000187:drug effects / Q000276:immunology; D010587:Phagocytosis / Q000187:drug effects*; D029424:Pulmonary Disease, Chronic Obstructive / Q000503:physiopathology; D015398:Signal Transduction / Q000187:drug effects; D012907:Smoking / Q000276:immunology / Q000503:physiopathology*; D014409:Tumor Necrosis Factor-alpha; D020741:rho GTP-Binding Proteins / Q000187:drug effects* / Q000378:metabolism; D054460:rho-Associated Kinases / Q000187:drug effects; D020742:rhoA GTP-Binding Protein
D014409:Tumor Necrosis Factor-alpha; D054460:rho-Associated Kinases; C526380:RhoA protein, mouse; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein