Hemodynamic regulation of perivalvular endothelial gene expression prevents deep venous thrombosis
John D. Welsh, … , Juan M. Jimenez, Mark L. Kahn
John D. Welsh, … , Juan M. Jimenez, Mark L. Kahn
Published December 2, 2019; First published November 11, 2019
Citation Information: J Clin Invest. 2019;129(12):5489-5500. https://doi.org/10.1172/JCI124791.
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Research Article Vascular biology

Hemodynamic regulation of perivalvular endothelial gene expression prevents deep venous thrombosis

Abstract

Deep venous thrombosis (DVT) and secondary pulmonary embolism cause approximately 100,000 deaths per year in the United States. Physical immobility is the most significant risk factor for DVT, but a molecular and cellular basis for this link has not been defined. We found that the endothelial cells surrounding the venous valve, where DVTs originate, express high levels of FOXC2 and PROX1, transcription factors known to be activated by oscillatory shear stress. The perivalvular venous endothelial cells exhibited a powerful antithrombotic phenotype characterized by low levels of the prothrombotic proteins vWF, P-selectin, and ICAM1 and high levels of the antithrombotic proteins thrombomodulin (THBD), endothelial protein C receptor (EPCR), and tissue factor pathway inhibitor (TFPI). The perivalvular antithrombotic phenotype was lost following genetic deletion of FOXC2 or femoral artery ligation to reduce venous flow in mice, and at the site of origin of human DVT associated with fatal pulmonary embolism. Oscillatory blood flow was detected at perivalvular sites in human veins following muscular activity, but not in the immobile state or after activation of an intermittent compression device designed to prevent DVT. These findings support a mechanism of DVT pathogenesis in which loss of muscular activity results in loss of oscillatory shear–dependent transcriptional and antithrombotic phenotypes in perivalvular venous endothelial cells, and suggest that prevention of DVT and pulmonary embolism may be improved by mechanical devices specifically designed to restore perivalvular oscillatory flow.

Authors

John D. Welsh, Mark H. Hoofnagle, Sharika Bamezai, Michael Oxendine, Lillian Lim, Joshua D. Hall, Jisheng Yang, Susan Schultz, James Douglas Engel, Tsutomu Kume, Guillermo Oliver, Juan M. Jimenez, Mark L. Kahn

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Figure 2

Perivalvular venous endothelium is resistant to leukocyte rolling and thrombus formation.

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Perivalvular venous endothelium is resistant to leukocyte rolling and th...
(A) Spontaneous leukocyte rolling at the saphenous venous valve was observed and quantitated in PROX1-GFP mice using rhodamine 6G. Arrow indicates the direction of flow, and arrowheads indicate individual leukocytes adherent to the vessel wall. Dashed lines outline the valve sinus region. The number of rolling leukocytes per millimeter vessel length was measured upstream of the valve (US), in the valve and valve sinus regions (V/S), and downstream of the valve (DS) (right) (n = 5 valves imaged in 3 mice). (B) Thrombus formation was stimulated in PROX1-GFP animals around the saphenous venous valve by application of extravascular thrombin, and clot visualized by accumulation of rhodamine 6G–positive platelets and leukocytes (orange) at 120 seconds (left). The percentage of vessel area covered by thrombus was measured upstream of the valve (US), in the valve and valve sinus regions (V/S), and downstream of the valve (DS) (right) (n = 8 valves imaged in 4 mice). (C) Visualization of thrombus formation over the saphenous venous valve of a PROX1-GFP mouse using anti-fibrin antibodies 10 minutes after thrombin application. The images are representative of those obtained from analysis of 3 separate experiments. White dashed lines indicate luminal venous endothelial cells, and green dashed lines indicate perivalvular endothelial cells. For each graph the mean is shown as the value of the bar with dots representing each replicate, and error bars indicate SD. Significance was determined by a paired 2-tailed t test and corrected for multiple comparisons. *P < 0.025; **P < 0.01.