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

EBioMedicine. 2019 Aug:46:274-289. doi: 10.1016/j.ebiom.2019.07.072. Epub 2019 Aug 5.

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.

Keywords: Cyclins; Fn14; Proliferation; Restenosis; TWEAK.

MeSH terms

  • Angioplasty / adverse effects*
  • Animals
  • Biomarkers
  • Cell Movement
  • Cell Proliferation
  • Coronary Restenosis / etiology*
  • Coronary Restenosis / metabolism*
  • Coronary Restenosis / 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
  • Tunica Intima / pathology

Substances

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