title: Nonproteolytic properties of murine alternatively spliced tissue factor: implications for integrin-mediated signaling in murine models.
abstract: This study was performed to determine whether murine alternatively spliced tissue factor (masTF) acts analogously to human alternatively spliced tissue factor (hasTF) in promoting neovascularization via integrin ligation. Immunohistochemical evaluation of a spontaneous murine pancreatic ductal adenocarcinoma model revealed increased levels of masTF and murine full-length tissue factor (mflTF) in tumor lesions compared with benign pancreas; furthermore, masTF colocalized with mflTF in spontaneous aortic plaques of Ldlr(-/-) mice, indicating that masTF is likely involved in atherogenesis and tumorigenesis. Recombinant masTF was used to perform in vitro and ex vivo studies examining its integrin-mediated biologic activity. Murine endothelial cells (ECs) rapidly adhered to masTF in a β3-dependent fashion. Using adult and embryonic murine ECs, masTF potentiated cell migration in transwell assays. Scratch assays were performed using murine and primary human ECs; the effects of masTF and hasTF were comparable in murine ECs, but in human ECs, the effects of hasTF were more pronounced. In aortic sprouting assays, the potency of masTF-triggered vessel growth was undistinguishable from that observed with hasTF. The proangiogenic effects of masTF were found to be Ccl2-mediated, yet independent of vascular endothelial growth factor. In murine ECs, masTF and hasTF upregulated genes involved in inflammatory responses; murine and human ECs stimulated with masTF and hasTF exhibited increased interaction with murine monocytic cells under orbital shear. We propose that masTF is a functional homolog of hasTF, exerting some of its key effects via β3 integrins. Our findings have implications for the development of murine models to examine the interplay between blood coagulation, atherosclerosis and cancer.
authors.affiliation: Department of Internal Medicine, Division of Hematology/Oncology, University of Cincinnati College of Medicine, Cincinnati, Ohio, United States of America.
authors.forename: Richard C
authors.initials: RC
authors.lastname: Godby
authors.forename: Yascha W
authors.initials: YW
authors.lastname: Van Den Berg
authors.forename: Ramprasad
authors.initials: R
authors.lastname: Srinivasan
authors.forename: Robert
authors.initials: R
authors.lastname: Sturm
authors.forename: David Y
authors.initials: DY
authors.lastname: Hui
authors.forename: Stephen F
authors.initials: SF
authors.lastname: Konieczny
authors.forename: Bruce J
authors.initials: BJ
authors.lastname: Aronow
authors.forename: Evgeny
authors.initials: E
authors.lastname: Ozhegov
authors.forename: Wolfram
authors.initials: W
authors.lastname: Ruf
authors.forename: Henri H
authors.initials: HH
authors.lastname: Versteeg
authors.forename: Vladimir Y
authors.initials: VY
authors.lastname: Bogdanov
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D002465:Cell Movement / Q000235:genetics; D016000:Cluster Analysis; D042783:Endothelial Cells / Q000378:metabolism; D015870:Gene Expression; D020869:Gene Expression Profiling; D016023:Integrins / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009000:Monocytes / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000235:genetics; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism; D058226:Plaque, Atherosclerotic / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D015398:Signal Transduction; D013925:Thromboplastin / Q000235:genetics* / Q000378:metabolism*
chemical_list: D016023:Integrins; D013925:Thromboplastin