TWEAK-Fn14 interaction enhances plasminogen activator inhibitor 1 and tissue factor expression in atherosclerotic plaques and in cultured vascular smooth muscle cells.
OBJECTIVE
atherosclerotic plaque development can conclude with a thrombotic acute event triggered by plaque rupture/erosion. Tumour necrosis factor-like weak inducer of apoptosis (TWEAK) is a member of the tumour necrosis factor superfamily that, through its receptor, fibroblast growth factor-inducible 14 (Fn14), participates in vascular remodelling, increasing vascular inflammatory responses and atherosclerotic lesion size in ApoE knockout mice. However, the role of the TWEAK-Fn14 axis in thrombosis has not been previously investigated.


RESULTS
we have examined whether TWEAK regulates expression of prothrombotic factors such as tissue factor (TF) and plasminogen activator inhibitor 1 (PAI-1) in atherosclerotic plaques as well as in human aortic vascular smooth muscle cells (hASMCs) in culture. Expression of TF and PAI-1 was colocalized and positively correlated with Fn14 in human carotid atherosclerotic plaques. In vitro, TWEAK increased TF and PAI-1 mRNA, protein expression and activity in hASMCs. All these effects were reversed using blocking anti-TWEAK monoclonal antibody, anti-Fn14 antibody or Fn14 small interfering RNA, indicating that TWEAK increased the prothrombotic state through its receptor, Fn14. Finally, ApoE(-/-) mice were fed a hyperlipidaemic diet for 10 weeks, then randomized and treated with saline (controls), TWEAK (10 microg/kg/day), anti-TWEAK neutralizing monoclonal antibody (1000 µg/kg/day), or non-specific immunoglobulin G (1000 microg/kg/day) daily for 9 days. Systemic TWEAK injection increased TF and PAI-1 protein expression in the aortic root of ApoE(-/-) mice. Conversely, TWEAK blocking antibodies diminished both TF and PAI-1 protein expression compared with non-specific immunoglobulin G-treated mice.


CONCLUSIONS
our results indicate that the TWEAK-Fn14 axis can regulate activation of TF and PAI-1 expression in vascular cells. TWEAK-Fn14 may be a therapeutic target in the prothrombotic complications associated with atherosclerosis.
Muñoz-García
Begoña
B
Renal and Vascular Research Laboratory, Instituto de Investigación Sanitaria, Fundación Jiménez Díaz , Autonoma University, Avd Reyes Católicos 2, 28040 Madrid, Spain.
Madrigal-Matute
Julio
J
Moreno
Juan A
JA
Martin-Ventura
Jose L
JL
López-Franco
Oscar
O
Sastre
Cristina
C
Ortega
Luis
L
Burkly
Linda C
LC
Egido
Jesús
J
Blanco-Colio
Luis M
LM
D000368:Aged; D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D002478:Cells, Cultured; D000074049:Cytokine TWEAK; D005260:Female; D015870:Gene Expression; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008875:Middle Aged; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism*; D058226:Plaque, Atherosclerotic / Q000235:genetics / Q000378:metabolism*; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D018124:Receptors, Tumor Necrosis Factor / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology; D058258:Serpin E2 / Q000235:genetics / Q000378:metabolism*; D000074050:TWEAK Receptor; D013925:Thromboplastin / Q000235:genetics / Q000378:metabolism*; D000079424:Tumor Necrosis Factor Inhibitors; D048069:Tumor Necrosis Factors / Q000235:genetics / Q000276:immunology / Q000378:metabolism* / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D001057:Apolipoproteins E; D000074049:Cytokine TWEAK; D017395:Plasminogen Activator Inhibitor 1; D034741:RNA, Small Interfering; D018124:Receptors, Tumor Necrosis Factor; D011994:Recombinant Proteins; C051321:SERPINE1 protein, human; D058258:Serpin E2; C511869:Serpine2 protein, mouse; C000613297:TNFRSF12A protein, human; C109743:TNFSF12 protein, human; D000074050:TWEAK Receptor; C000613299:Tnfrsf12a protein, mouse; C491645:Tnfsf12 protein, mouse; D000079424:Tumor Necrosis Factor Inhibitors; D048069:Tumor Necrosis Factors; D013925:Thromboplastin