MIF tautomerase inhibitor TE-11 prevents inflammatory macrophage activation and glycolytic reprogramming while reducing leukocyte migration and improving Crohn's disease-like colitis in male mice

Eszter Vámos ¹ Viola Bagóné Vántus ¹ Péter Deák ¹ Nikoletta Kálmán ¹ Eva Maria Sturm ² Barsha Baisakhi Nayak ² Lilla Makszin ³ Tamás Loránd ¹ Ferenc Gallyas ¹ Balázs Radnai ^1*

• ¹ Department of Biochemistry and Medical Chemistry, Medical School, University of Pécs, Pécs, Hungary
• ² Otto-Loewi Research Center for Vascular Biology, Immunology and Inflammation, Division of Pharmacololgy, Medical University of Graz, Graz, Austria
• ³ Institute of Bioanalysis, Medical School, Szentágothai Research Center, University of Pécs, Pécs, Hungary

Background & aims: Crohn's disease (CD) is a chronic inflammatory disorder primarily affecting the gastrointestinal tract. Leukocyte recruitment, M1 macrophage polarization and associated metabolic reprogramming are hallmarks of its pathomechanism. Here, we tested TE-11, a potent MIF tautomerase inhibitor (IC50=5.63 μmol/dm 3 ) in experimental Crohn's disease in male mice, in leukocyte recruitment and in inflammatory M1 macrophage activation.Methods: 2,4,6-trinitrobenzenesulfonic acid-(TNBS)-induced colitis was utilized as a CDmodel in male mice. We performed macroscopic scoring and cytokine measurements. We also analyzed MIF-induced leukocyte migration and evaluated apoptosis. LPS+IFN-γ-induced RAW264.7 cells were applied as a M1 macrophage model. We performed qPCR, ROS and nitrite determinations, ELISA measurements, mitochondrial oxygen consumption rate and extracellular acidification rate determinations.: TE-11 improved mucosal damage, reduced inflammation score and concentration of IL-1β and IL-6 in the colon. It inhibited MIF-induced human blood eosinophil and neutrophil migration and counteracted the anti-apoptotic effect of MIF. In macrophages, MIF inhibition prevented M1 polarization by downregulating HIF-1α gene expression in LPS+IFN-γ-activated cells. Additionally, the molecule reduced mRNA transcription and protein expression of chemokine CCL-2 and cytokine IL-6 while further increasing SOD2 gene transcription and decreased ROS and nitrite production in macrophages. During inflammatory metabolic reprogramming, TE-11 prevented LPS+IFN-γ-induced metabolic shift from OXPHOS to glycolysis. Similarly to anti-inflammatory M2 cells, TE-11 improved mitochondrial energy production by increasing basal respiration, ATP production, coupling efficiency, maximal respiration and spare respiratory capacity. Conclusion: Comprehensively, TE-11, a MIF tautomerase inhibitor ameliorates CD-like colitis, reduces MIF-induced eosinophil and neutrophil migration and prevents M1 polarization and associated metabolic reprogramming; therefore, it may prove beneficial as a potential drug candidate regarding CD therapy.