Direct Thrombin Inhibitor Dabigatran Compromises Pulmonary Endothelial Integrity in a Murine Model of Breast Cancer Metastasis to the Lungs; the Role of Platelets and Inflammation-Associated Haemostasis

Abstract

Activation of the coagulation cascade favours metastatic spread, but antithrombotic therapy might also have detrimental effects on cancer progression. In this study, we characterized the effects of dabigatran, a direct reversible thrombin inhibitor, on the pulmonary endothelial barrier and metastatic spread in a murine model of breast cancer metastasis. Dabigatran etexilate (100 mg kg−1) was administered to mice twice daily by oral gavage. Pulmonary metastasis, pulmonary endothelium permeability in vivo, and platelet reactivity were evaluated after intravenous injection of 4T1 breast cancer cells into BALB/c mice. The effect of dabigatran on platelet-dependent protection of pulmonary endothelial barrier in the presence of an inflammatory stimulus was also verified in vitro using human lung microvascular endothelial cell (HLMVEC) cultures. Dabigatran-treated mice harbored more metastases in their lungs and displayed increased pulmonary endothelium permeability after cancer cell injection. It was not associated with altered lung fibrin deposition, changes in INFγ, or complement activation. In the in vitro model of the pulmonary endothelial barrier, dabigatran inhibited platelet-mediated protection of pulmonary endothelium. In a murine model of breast cancer metastasis, dabigatran treatment promoted pulmonary metastasis by the inhibition of platelet-dependent protection of pulmonary endothelial barrier integrity.

Platelets in COVID-19 disease: friend, foe, or both?

“…Freshly transfused platelets attenuated lung injury in the murine model of extracorporeal circulation (ECC)-induced systemic inflammatory response [ 23 ] and this protective effect of freshly transfused platelets was impaired in the presence of an antiplatelet drug, tirofiban. A similar effect was also observed in vitro, whereby the protective effect of thrombin-induced platelet releasate on the human microvascular lung endothelial barrier was significantly attenuated in the presence of the thrombin inhibitor dabigatran [ 168 ]. Moreover, via docking to the von Willebrand factor, platelets could prevent the leaks associated with leukocyte transmigration by releasing Angpt-1, which activates endothelial receptor Tie-2 signaling [ 169 ].…”

“…Activation of platelet CLEC-2 [172] and GPVI [173] receptors initiates intra-platelet ITAM-dependent signaling that results in protective effects on the endothelium [173]. Finally, thrombin-induced pEVs protect endothelial barrier integrity [174] thrombin-induced platelet releasate on the human microvascular lung endothelial barrier was significantly attenuated in the presence of the thrombin inhibitor dabigatran [168]. Moreover, via docking to the von Willebrand factor, platelets could prevent the leaks associated with leukocyte transmigration by releasing Angpt-1, which activates endothelial receptor Tie-2 signaling [169].…”

Endothelial-mesenchymal transition induced by metastatic 4T1 breast cancer cells in pulmonary endothelium in aged mice

“…The plasma concentration of a panel of endothelial dysfunction biomarkers was quantified using microLC/MS-MRM method as described previously ( Walczak et al, 2015 ; Suraj et al, 2018 ; Suraj et al, 2019a ; Suraj et al, 2019b ; Smeda et al, 2022 ) and a UPLC Nexera system (Shimadzu; Kyoto, Japan) connected to a highly sensitive QTrap 5500 mass spectrometer (Sciex; Framingham, MA, United States). The panel included eighteen glycocalyx disruption biomarkers: syndecan-1 (SDC-1) and endocan (ESM-1); hemostasis: von Willebrand factor (vWF), tissue plasminogen activator (t-PA), plasminogen activator inhibitor 1 (PAI-1), and thrombin activatable fibrinolysis inhibitor (TAFI); endothelium inflammation: soluble vascular cell adhesion molecule 1 (sVCAM-1), soluble intercellular adhesion molecule 1 (sICAM-1), and soluble form of E-selectin (sE-sel); platelet activation: soluble form of P-selectin (sP-sel) and thrombospondin 1 (THBS-1); endothelium permeability: angiopoietin 1 (Ang-1), angiopoietin 2 (Ang-2), soluble form of FMS-like tyrosine kinase 1 (sFLT-1), and soluble form of Tie-2 receptor (sTie-2); other proteins and peptides related to endothelial function: adrenomedullin (ADM), adiponectin (ADN) and annexin V (ANXA5).…”

Oleanolic acid inhibits the tumor progression by regulating Lactobacillus through the cytokine-cytokine receptor interaction pathway in 4T1-induced mice breast cancer model

“…Activation of the coagulation cascade favors the spread of breast cancer metastasis [ 58 ], and Clu overexpression in breast cancer in the complement and coagulation cascades pathway is associated with lymphatic metastasis [ 54 ], however, under OA therapy, Clu downregulation may block the coagulation cascade pathway, thereby affecting the progression of breast cancer. As part of the complement system, Hc responds when the body is injured and tissue cells are damaged, which is an important part of the host.…”