Tu1683 Expression of Thymidine Phosphorylase in Ulcerative Colitis and Efficacy of Its Specific Inhibitor for the Treatment of Ulcerative Colitis

Abstract

protein were collected. Cytokine levels in cell supernatants were determined using the Bioplex 3D suspension array system (Bio-Rad). Preadipocyte supernatants were also applied on cultures of human colonic NCM-460 cells for 24hs and RNA and protein were collected. Inflammatory molecule mRNA expression and activation of intracellular kinases were evaluated with Luminex inflammatory panels (FlexScript LDA), and phospho-protein multiplex panels, respectively. Results: Preadipocytes isolated from IBD patients demonstrated differential mediator secretion patterns compared to preadipocytes from control and obese patients. Of the 42 proinflammatory cytokines and growth factors tested, IL-7/IL-8/Gro/MCP3/VEGF/ PDGF-AA showed statistically significant differences between control, obese and IBD preadipocytes (p ,0.05). Notably, there were significant differences in mediator secretion between preadipocytes isolated from UC and CD patients, in particular VEGF and PDGF (p , 0.01 for both). NCM-460 cells had distinct GSK-3, AKT kinase activation (p , 0.05 for both) and cytokine mRNA expression patterns of TNF, IL-2, IL-3, IL-17, VEGF, MIP-1 β, MIP-3, RANTES (p,0.05) following exposure to preadipocyte supernatants from all three pathological conditions compared to controls. Conclusions: Our data suggest the existence of preadipocyte-disease-dependent mediators contributing to the generation of differential colonocyte responses. Together with reports indicating altered levels of several adipokines during IBD, our results provide the first direct evidence for the importance of adipose tissue homeostasis during obesity in determining the outcome of IBD. Supported by NIH P50 DK064539 (CP & IK), DK047343, DK060729 (CP), The Broad Foundation (BMRP) (IK, RA), and the Blinder Research Foundation for Crohn's Disease (AS).