Experimental metastasis and primary tumor growth in mice with hemophilia A.
During experimental lung metastasis, tumor cells adhere to the pulmonary microvasculature and activate coagulation via surface-expressed tissue factor (TF), leading to local fibrin deposition and platelet aggregation. While interventional studies have demonstrated great efficacy of anticoagulants and antiplatelet agents in inhibiting metastasis, no information is available on how tumor biology may be affected by congenital bleeding disorders such as hemophilia A. We therefore used a syngeneic model to study experimental metastasis and primary tumor growth in factor VIII (FVIII)-deficient mice. By conventional reverse transcription-polymerase chain reaction, flow cytometry, and one-stage clotting assays, we demonstrated constitutive expression of TF mRNA, antigen, and procoagulant activity in the murine B16F10 melanoma cell line. In hemophilic mice, B16F10 lung metastasis was significantly (P < 0.001) enhanced by a single dose of human FVIII (100 U kg(-1)), suggesting that FVIII played a critical role during the early blood-borne phase of the metastatic cascade. In contrast, lung seeding was significantly (P < 0.05) reduced by lepirudin, a direct thrombin inhibitor, suggesting that thrombin generation contributed to pulmonary metastasis even in the absence of FVIII. Consistent with this finding, intravenous injection of B16F10 cell-evoked laboratory changes of a hemolytic thrombotic microangiopathy and consumptive coagulopathy in both hemophilic and non-hemophilic mice. Subcutaneous implantation of B16F10 cells into mice with hemophilia A gave rise to primary tumors in an exponential growth pattern similar to that observed in non-hemophilic mice. Although TF expression by B16F10 cells may promote thrombin-dependent metastasis in mice with hemophilia A, amplification of coagulation by host FVIII appears to be necessary for maximum lung seeding.
Langer
F
F
II. Medizinische Klinik und Poliklinik, Universitätsklinikum Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany. langer@uke.uni-hamburg.de
Amirkhosravi
A
A
Ingersoll
S B
SB
Walker
J M
JM
Spath
B
B
Eifrig
B
B
Bokemeyer
C
C
Francis
J L
JL
D000818:Animals; D002455:Cell Division; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D006467:Hemophilia A / Q000473:pathology*; D008297:Male; D051379:Mice; D009362:Neoplasm Metastasis; D020133:Reverse Transcriptase Polymerase Chain Reaction