Pulmonary arterial hypertension (PAH) is a poorly characterized disease of unclear etiology that affects individuals of all ages. Vascular remodeling and increase in pulmonary artery (PA) and right ventricle (RV) pressures are two major culprits in RV failure and death in PAH. Recent advances in the study of PAH suggest that endothelial cell proliferation is an early instigator of this hallmark remodeling. We postulated that Axl receptor tyrosine kinase (implicated in pro-proliferative and pro-survival signaling in cancerous cells) could mediate endothelial proliferation and thus hemodynamic changes occurring in PAH. Using immunofluorescent microscopy of lung microvessels of human PAH vs. non-PAH, we observed Axl expression on intimal endothelial cells but not medial smooth muscle cells. Furthermore, digitized microscopy revealed that Axl tended to increase on the endothelium of PAH vessels (1.65±0.15-fold vs. non-PAH; n=3-4;
p=0.057
). To address the role of Axl
in vivo
, an Axl inhibitor R428 was employed in a mouse model of pulmonary hypertension. C57Bl/6 mice were subjected to hypoxia at pO
2
=10% and VEGF receptor antagonist SU5416 (Su/Ch) or normoxia (Norm) for 3 wks. Indeed, Su/Ch caused a significant rise in lung Axl protein and mRNA (7.1±0.4- and 2.4±0.5-fold, Su/Ch vs. Norm, protein and mRNA, respectively; n=3-6; p<0.01). As predicted, RV pressure (RVP) rose from 27±0.5 to 43±1.8 mmHg (Norm vs. Su/Ch; n=6; p<0.01). However, we did not observe a decrease in RVP with twice-daily gavage of 75 mg/kg R428 (43±1.4 mmHg, Su/Ch + R428; n=6). A similar pattern was observed with mean PA pressure (18.4±0.3 and 28.7±1.2 mmHg, Norm vs. Su/Ch, p<0.01; 28.7±0.9 mmHg, Su/Ch + R428), RV resistance (1403±256 vs. 2703±464 Wood units, Norm vs. Su/Ch, n/s; vs. 3610±625 Wood units, Su/Ch + R428) and Fulton index (0.26±0.01 and 0.34±0.02, Norm vs. Su/Ch, p<0.05; 0.38±0.02, Su/Ch + R428). In conclusion, our preliminary results demonstrate upregulated Axl expression in the endothelium of PAH patients and in lungs of PH mice and suggest that Axl kinase may play a novel role in pulmonary vascular endothelial proliferation and remodeling in PAH. It remains to be determined whether drug bioavailability or severity of disease precluded an ameliorative effect of an Axl inhibitor.
A Novel p38 Mitogen-activated Protein Kinase/Elk-1 Transcription Factor-dependent Molecular Mechanism Underlying Abnormal Endothelial Cell Proliferation in Plexogenic Pulmonary Arterial Hypertension
