Inhibition of lymphangiogenesis in vitro and in vivo by the multikinase inhibitor nintedanib

Abstract

PurposeTo investigate the feasibility of nintedanib, a novel triple angiokinase inhibitor, for inhibiting lymphatic endothelial cell (LEC)-induced lymphangiogenesis in vitro and inflammatory corneal lymphangiogenesis in vivo.Materials and methodsMethylthiazolyldiphenyl-tetrazolium bromide (MTT) test, transwell system, and tube-formation assay were used to evaluate the effects of nintedanib on the proliferation, migration, and tube formation of LECs stimulated by vascular endothelial growth factor-C (VEGF-C), basic fibroblast growth factor (bFGF), or platelet-derived growth factor-BB (PDGF-BB). The murine model of suture-induced corneal neovascularization was used to assess the anti-hemangiogenic and anti-lymphangiogenic effects of nintedanib via systemic and topical applications. Corneal flatmounts were stained with lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) and CD31, and the areas of involved blood and lymph vessels were analyzed morphometrically. Corneal cryosections were stained with F4/80 to evaluate inflammatory cell recruitment.ResultsWe observed a significant enhanced effect of LEC proliferation, migration, and tube formation with the administration of VEGF-C, PDGF-BB, and bFGF, respectively, which was diminished by nintedanib. Both topical and systemic applications of nintedanib inhibited suture-induced hemangiogenesis and lymphangiogenesis in the murine cornea. A reduction in F4/80+ cell infiltration was observed at day 14 after corneal suture for both systemic and topical applications of nintedanib. In comparison with controls, 61% of F4/80+ cell recruitment was inhibited via the systemic application of nintedanib, while 49% of F4/80+ cell recruitment was inhibited with the topical application of nintedanib.ConclusionNintedanib was shown to inhibit in vitro lymphangiogenesis stimulated by VEGF-C, bFGF, and PDGF-BB. Applied topically or systemically, it effectively inhibited corneal hemangiogenesis and lymphangiogenesis, accompanied by reduced inflammatory cell recruitment, which represents a new promising treatment for graft rejection after penetrating keratoplasty.

Nintedanib attenuates peritoneal fibrosis by inhibiting mesothelial‐to‐mesenchymal transition, inflammation and angiogenesis

“…We cannot exclude the possibility that nintedanib may also suppress peritoneal fibrosis through inhibition of lymphangiogenesis. In this regard, it has been reported by Lin et al 34 that nintedanib inhibited suture‐induced corneal lymphangiogenesis and inflammatory cell recruitment in vivo and in vitro. Increased expression of VEGF‐C induced by TGF‐β can trigger lymphangiogenesis during peritoneal fibrosis, 35,36 and increased peritoneal lymphatic vessels can increase absorption of dialysate with high‐dose glucose during PD treatment, thereby reducing the ultrafiltration of peritoneal membranes 37 .…”

Nintedanib inhibits the development and progression of peritoneal fibrosis

“…Nevertheless, we cannot exclude the possibility that nintedanib may also suppress peritoneal fibrosis through inhibition of lymphangiogenesis. In this regard, Lin et al have reported that nintedanib inhibited suture-induced corneal lymphangiogenesis, accompanied by reduced inflammatory cell recruitment in vitro and in vitro [29]. During peritoneal fibrosis, TGF-β induced VEGF-C expression is able to trigger lymphangiogenesis [30,31]; increased peritoneal lymphatic vessels constantly absorb dialysate during PD treatment, which reduces effective ultrafiltration [32].…”

Advances in Drugs Targeting Lymphangiogenesis for Preventing Tumor Progression and Metastasis

“…Triple angiokinase inhibitor, Nintedanib, adequately dampened lymphangiogenesis stimulated by VEGF-C, bFGF, and PDGF-BB in a suture-induced corneal neovascularization assay. Both topical and systemic applications showed positive results, though the in vivo safety of topical use wasn’t tested yet ( 52 ). Interestingly, other than the clinical application in renal cell carcinoma, Axitinib significantly inhibited LEC proliferation and lymphangiogenesis in a study of allergic asthma ( 68 ).…”