Human Plasma Thioredoxin-80 Increases With Age and in ApoE−/− Mice Induces Inflammation, Angiogenesis, and Atherosclerosis

• Published in: Circulation (New York, N.Y.) Vol. 136; no. 5; pp. 464 - 475
• Main Authors: Couchie, Dominique, Vaisman, Boris, Abderrazak, Amna, Mahmood, Dler Faieeq Darweesh, Hamza, Magda M., Canesi, Fanny, Diderot, Vimala, El Hadri, Khadija, Nègre-Salvayre, Anne, Le Page, Aurélie, Fulop, Tamas, Remaley, Alan T., Rouis, Mustapha
• Format: Journal Article
• Language: English
• Published: United States by the American College of Cardiology Foundation and the American Heart Association, Inc 01.08.2017; American Heart Association
• Subjects: ADAM10 Protein - metabolism; ADAM17 Protein - metabolism; Adult; Aged; Aging; Animals; Apolipoproteins E - deficiency; Apolipoproteins E - genetics; Atherosclerosis - metabolism; Atherosclerosis - pathology; Biomarkers - blood; Biomarkers - metabolism; Disease Models, Animal; Down-Regulation - drug effects; Enzyme-Linked Immunosorbent Assay; Female; Human health and pathology; Humans; Immunohistochemistry; Inflammation; Interleukin-18 - blood; Interleukin-1beta - blood; Leukocytes, Mononuclear - metabolism; Life Sciences; Lipopolysaccharides - toxicity; Macrophages - cytology; Macrophages - drug effects; Macrophages - metabolism; Male; Mechanistic Target of Rapamycin Complex 1; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Microscopy, Confocal; Multiprotein Complexes - antagonists & inhibitors; Multiprotein Complexes - metabolism; Neovascularization, Physiologic - drug effects; NLR Family, Pyrin Domain-Containing 3 Protein - metabolism; Peptide Fragments - blood; Peptide Fragments - genetics; Peptide Fragments - metabolism; Peptide Fragments - pharmacology; Proto-Oncogene Proteins c-akt - metabolism; Recombinant Proteins - biosynthesis; Recombinant Proteins - isolation & purification; Recombinant Proteins - pharmacology; Sirolimus - pharmacology; Thioredoxins - blood; Thioredoxins - genetics; Thioredoxins - metabolism; Thioredoxins - pharmacology; TOR Serine-Threonine Kinases - antagonists & inhibitors; TOR Serine-Threonine Kinases - metabolism; atherosclerosis; macrophages; inflammasomes; inflammation; neovascularization; thioredoxins; interleukin-1beta
• ISSN: 0009-7322; 1524-4539; 1524-4539
• DOI: 10.1161/CIRCULATIONAHA.117.027612

BACKGROUND:Thioredoxin (TRX)-1, a ubiquitous 12-kDa protein, exerts antioxidant and anti-inflammatory effects. In contrast, the truncated form, called TRX80, produced by macrophages induces upregulation of proinflammatory cytokines. TRX80 also promotes the differentiation of mouse peritoneal and human macrophages toward a proinflammatory M1 phenotype. METHODS:TRX1 and TRX80 plasma levels were determined with a specific ELISA. A disintegrin and metalloproteinase domain-containing protein (ADAM)-10, ADAM-17, and ADAM-10 activities were measured with SensoLyte 520 ADAM10 Activity Assay Kit, Fluorimetric, and InnoZyme TACE Activity Kit, respectively. Western immunoblots were performed with specific antibodies to ADAM-10 or ADAM-17. Angiogenesis study was evaluated in vitro with human microvascular endothelial cells-1 and in vivo with the Matrigel plug angiogenesis assay in mice. The expression of macrophage phenotype markers was investigated with real-time polymerase chain reaction. Phosphorylation of Akt, mechanistic target of rapamycin, and 70S6K was determined with specific antibodies. The effect of TRX80 on NLRP3 inflammasome activity was evaluated by measuring the level of interleukin-1β and -18 in the supernatants of activated macrophages with ELISA. Hearts were used for lesion surface evaluation and immunohistochemical studies, and whole descending aorta were stained with Oil Red O. For transgenic mice generation, the human scavenger receptor (SR-A) promoter/enhancer was used to drive macrophage-specific expression of human TRX80 in mice. RESULTS:In this study, we observed a significant increase of plasma levels of TRX80 in old subjects compared with healthy young subjects. In parallel, an increase in expression and activity of ADAM-10 and ADAM-17 in old peripheral blood mononuclear cells compared with those of young subjects was observed. Furthermore, TRX80 was found to colocalize with tumor necrosis factor-α, a macrophage M1 marker, in human atherosclerotic plaque. In addition, TRX80 induced the expression of murine M1 macrophage markers through Akt2/mechanistic target of rapamycin–C1/70S6K pathway and activated the inflammasome NLRP3, leading to the release of interleukin-1β and -18, potent atherogenic cytokines. Moreover, TRX80 exerts a powerful angiogenic effect in both in vitro and in vivo mouse studies. Finally, transgenic mice that overexpress human TRX80 specifically in macrophages of apoE mice have a significant increase of aortic atherosclerotic lesions. CONCLUSIONS:TRX80 showed an age-dependent increase in human plasma. In mouse models, TRX80 was associated with a proinflammatory status and increased atherosclerosis.