title: TWEAK favors phosphate-induced calcification of vascular smooth muscle cells through canonical and non-canonical activation of NFκB.
abstract: Vascular calcification (VC) is associated with increased cardiovascular mortality in aging, chronic kidney disease (CKD), type 2 diabetes mellitus (T2DM) and atherosclerosis. TNF-like weak inducer of apoptosis (TWEAK) recently emerged as a new biomarker for the diagnosis and prognosis of cardiovascular diseases. TWEAK binding to its functional receptor Fn14 was reported to promote several steps of atherosclerotic plaque progression. However, no information is currently available on the role of TWEAK/Fn14 on the development of medial calcification, which is highly prevalent in aging, CKD and T2DM. This study explored the involvement of TWEAK in human vascular smooth muscle cells (h-VSMCs) calcification in vitro. We report that TWEAK binding to Fn14 promotes inorganic phosphate-induced h-VSMCs calcification, favors h-VSMCs osteogenic transition, decreasing acta2 and myh11 and increasing bmp2 mRNA and tissue non-specific alkaline phosphatase (TNAP), and increases MMP9 activity. Blockade of the canonical NFκB pathway reduced by 80% TWEAK pro-calcific properties and decreased osteogenic transition, TNAP and MMP9 activity. Blockade of non-canonical NFκB signaling by a siRNA targeting RelB reduced by 20% TWEAK pro-calcific effects and decreased TWEAK-induced loss of h-VSMCs contractile phenotype and MMP9 activity, without modulating bmp2 mRNA or TNAP activity. Inhibition of ERK1/2 activation by a MAPK kinase inhibitor did not influence TWEAK pro-calcific properties. Our results suggest that TWEAK/Fn14 directly favors inorganic phosphate-induced h-VSMCs calcification by activation of both canonical and non-canonical NFκB pathways. Given the availability of neutralizing anti-TWEAK strategies, our study sheds light on the TWEAK/Fn14 axis as a novel therapeutic target in the prevention of VC.
authors.affiliation: Nephrology, IIS-Fundación Jiménez Díaz, School of Medicine, Universidad Autónoma de Madrid, Madrid 28040, Spain.
authors.forename: L
authors.initials: L
authors.lastname: Hénaut
authors.affiliation: Nephrology, IIS-Fundación Jiménez Díaz, School of Medicine, Universidad Autónoma de Madrid, Madrid 28040, Spain.|REDINREN, Madrid 28040, Spain.
authors.forename: A B
authors.initials: AB
authors.lastname: Sanz
authors.affiliation: Nephrology, IIS-Fundación Jiménez Díaz, School of Medicine, Universidad Autónoma de Madrid, Madrid 28040, Spain.
authors.forename: D
authors.initials: D
authors.lastname: Martin-Sanchez
authors.affiliation: Nephrology, IIS-Fundación Jiménez Díaz, School of Medicine, Universidad Autónoma de Madrid, Madrid 28040, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Carrasco
authors.affiliation: Nephrology, IIS-Fundación Jiménez Díaz, School of Medicine, Universidad Autónoma de Madrid, Madrid 28040, Spain.
authors.forename: R
authors.initials: R
authors.lastname: Villa-Bellosta
authors.affiliation: Cardiac Surgery, IIS-Fundación Jiménez Díaz, School of Medicine, Universidad Autónoma de Madrid, Madrid 28040, Spain.
authors.forename: G
authors.initials: G
authors.lastname: Aldamiz-Echevarria
authors.affiliation: Division of Nephrology, Ambroise Paré University Hospital, APHP, University of Paris Ouest -Versailles-Saint-Quentin-en-Yvelines (UVSQ), Boulogne-Billancourt, Paris, France.|INSERM U1018, Research Centre in Epidemiology and Population Health (CESP) Team 5, Villejuif, Paris, France.
authors.forename: Z A
authors.initials: ZA
authors.lastname: Massy
authors.affiliation: Nephrology, IIS-Fundación Jiménez Díaz, School of Medicine, Universidad Autónoma de Madrid, Madrid 28040, Spain.|REDINREN, Madrid 28040, Spain.
authors.forename: M D
authors.initials: MD
authors.lastname: Sanchez-Nino
authors.affiliation: Nephrology, IIS-Fundación Jiménez Díaz, School of Medicine, Universidad Autónoma de Madrid, Madrid 28040, Spain.|REDINREN, Madrid 28040, Spain.|Fundación Renal Iñigo Alvarez de Toledo-IRSIN, Madrid 28040, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Ortiz
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism; D000074049:Cytokine TWEAK; D005786:Gene Expression Regulation; D006801:Humans; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000235:genetics / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000235:genetics / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism / Q000494:pharmacology*; D061251:Primary Cell Culture; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D018124:Receptors, Tumor Necrosis Factor / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D000074050:TWEAK Receptor; D051841:Transcription Factor RelB / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D048069:Tumor Necrosis Factors / Q000235:genetics* / Q000378:metabolism; D061205:Vascular Calcification / Q000235:genetics / Q000378:metabolism / Q000473:pathology
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D000074049:Cytokine TWEAK; C509903:MYH11 protein, human; D010710:Phosphates; C497593:RELB protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D018124:Receptors, Tumor Necrosis Factor; C000613297:TNFRSF12A protein, human; C109743:TNFSF12 protein, human; D000074050:TWEAK Receptor; D048069:Tumor Necrosis Factors; D051841:Transcription Factor RelB; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; C548267:ALPL protein, human; D000469:Alkaline Phosphatase; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9; D018995:Myosin Heavy Chains