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

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

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)274-289
Number of pages16
JournalEBioMedicine
Volume46
DOIs
StatePublished - Aug 2019

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Keywords

  • Cyclins
  • Fn14
  • Proliferation
  • Restenosis
  • TWEAK

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Méndez-Barbero, N., Gutierrez-Muñoz, C., Madrigal-Matute, J., Mínguez, P., Egido, J., Michel, J. B., Martín-Ventura, J. L., Esteban, V., & Blanco-Colio, L. M. (2019). A major role of TWEAK/Fn14 axis as a therapeutic target for post-angioplasty restenosis. EBioMedicine, 46, 274-289. https://doi.org/10.1016/j.ebiom.2019.07.072