A major role of TWEAK/Fn14 axis as a therapeutic target for post-angioplasty restenosis.

Abstract

BACKGROUND:

Tumor necrosis factor-like weak inducer of apoptosis (Tnfsf12; TWEAK) and its receptor Fibroblast growth factor-inducible 14 (Tnfrsf12a; Fn14) participate in the inflammatory response associated with vascular remodeling. However, the functional effect of TWEAK on vascular smooth muscle cells (VSMCs) is not completely elucidated.

METHODS:

Next generation sequencing-based methods were performed to identify genes and pathways regulated by TWEAK in VSMCs. Flow-citometry, wound-healing scratch experiments and transwell migration assays were used to analyze VSMCs proliferation and migration. Mouse wire injury model was done to evaluate the role of TWEAK/Fn14 during neointimal hyperplasia.

FINDINGS:

TWEAK up-regulated 1611 and down-regulated 1091 genes in VSMCs. Using a gene-set enrichment method, we found a functional module involved in cell proliferation defined as the minimal network connecting top TWEAK up-regulated genes. In vitro experiments in wild-type or Tnfrsf12a deficient VSMCs demonstrated that TWEAK increased cell proliferation, VSMCs motility and migration. Mechanistically, TWEAK increased cyclins (cyclinD1), cyclin-dependent kinases (CDK4, CDK6) and decreased cyclin-dependent kinase inhibitors (p15lNK4B) mRNA and protein expression. Downregulation of p15INK4B induced by TWEAK was mediated by mitogen-activated protein kinase ERK and Akt activation. Tnfrsf12a or Tnfsf12 genetic depletion and pharmacological intervention with TWEAK blocking antibody reduced neointimal formation, decreasing cell proliferation, cyclin D1 and CDK4/6 expression, and increasing p15INK4B expression compared with wild type or IgG-treated mice in wire-injured femoral arteries. Finally, immunohistochemistry in human coronary arteries with stenosis or in-stent restenosis revealed high levels of Fn14, TWEAK and PCNA in VSMCs enriched areas of the neointima as compared with healthy coronary arteries.

INTERPRETATION:

Our data define a major role of TWEAK/Fn14 in the control of VSMCs proliferation and migration during neointimal hyperplasia after wire injury in mice, and identify TWEAK/Fn14 as a potential target for treating in-stent restenosis. FUND: ISCiii-FEDER, CIBERCV and CIBERDEM.

• Authors: Nerea Méndez-Barbero, Carmen Gutierrez-Muñoz, Julio Madrigal-Matute, Pablo Mínguez, Jesús Egido, Jean-Baptiste Michel, Jose L Martín-Ventura, Vanesa Esteban, Luis M Blanco-Colio
• Journal: EBioMedicine (EBioMedicine) Vol. 46 Pg. 274-289 (Aug 2019) ISSN: 2352-3964 [Electronic] Netherlands
• PMID: 31395500 (Publication Type: Journal Article)
• Copyright: Copyright © 2019. Published by Elsevier B.V.

Chemical References

• Biomarkers
• Cytokine TWEAK
• TNFRSF12A protein, human
• TNFSF12 protein, human
• TWEAK Receptor

Topics

• Angioplasty (adverse effects)
• Animals
• Biomarkers
• Cell Movement
• Cell Proliferation
• Coronary Restenosis (etiology, metabolism, pathology)
• Cytokine TWEAK (metabolism)
• Disease Models, Animal
• Flow Cytometry
• Gene Expression Profiling
• Gene Expression Regulation
• Gene Regulatory Networks
• Immunohistochemistry
• Mice
• Mice, Knockout
• Models, Biological
• Myocytes, Smooth Muscle (metabolism)
• Postoperative Complications
• Signal Transduction (drug effects)
• TWEAK Receptor (metabolism)
• Tunica Intima (metabolism, pathology)