Nicotinamide phosphoribosyltransferase inhibition reduces intraplaque CXCL1 production and associated neutrophil infiltration in atherosclerotic mice

Summary

Pharmacological treatments targeting CXC chemokines and the associated neutrophil activation and recruitment into atherosclerotic plaques hold promise for treating cardiovascular disorders. Therefore, we investigated whether FK866, a nicotinamide phosphoribosyltransferase (NAMPT) inhibitor with anti-inflammatory properties that we recently found to reduce neutrophil recruitment into the ischaemic myocardium, would exert beneficial effects in a mouse atherosclerosis model. Atherosclerotic plaque formation was induced by carotid cast implantation in ApoE-/- mice that were fed with a Western-type diet. FK866 or vehicle were administrated intraperitoneally from week 8 until week 11 of the diet. Treatment with FK866 reduced neutrophil infiltration and MMP-9 content and increased collagen levels in atherosclerotic plaques compared to vehicle. No effect on other histological parameters, including intraplaque lipids or macrophages, was observed. These findings were associated with a reduction in both systemic and intraplaque CXCL1 levels in FK866-treated mice. In vitro, FK866 did not affect MMP-9 release by neutrophils, but it strongly reduced CXCL1 production by endothelial cells which, in the in vivo model, were identified as a main CXCL1 source at the plaque level. CXCL1 synthesis inhibition by FK866 appears to reflect interference with nuclear factor-κB signalling as shown by reduced p65 nuclear levels in endothelial cells pre-treated with FK866. In conclusion, pharmacological inhibition of NAMPT activity mitigates inflammation in atherosclerotic plaques by reducing CXCL1-mediated activities on neutrophils. These results support further assessments of NAMPT inhibitors for the potential prevention of plaque vulnerability.

Keywords

Carotid artery - inflammation - atherosclerosis

• References

• 1 Van Gool F, Gallí M, Gueydan C. et al. Intracellular NAD levels regulate tumor necrosis factor protein synthesis in a sirtuin-dependent manner. Nat Med 2009; 15: 206-210.
• 2 Busso N, Karababa M, Nobile M. et al. Pharmacological inhibition of nicotina-mide phosphoribosyltransferase/visfatin enzymatic activity identifies a new inflammatory pathway linked to NAD. PLoS One 2008; 3: e2267
• 3 Bruzzone S, Fruscione F, Morando S. et al. Catastrophic NAD+ depletion in activated T lymphocytes through NAMPT inhibition reduces demyelination and disability in EAE. PLoS One 2009; 4: e7897
• 4 Montecucco F, Bauer I, Braunersreuther V. et al. Inhibition of nicotinamide phosphoribosyltransferase reduces neutrophil-mediated injury in myocardial infarction. Antioxid Redox Signal 2013; 18: 630-641.
• 5 Hasmann M, Schemainda I. FK866, a highly specific noncompetitive inhibitor of nicotinamide phosphoribosyltransferase, represents a novel mechanism for induction of tumor cell apoptosis. Cancer Res 2003; 63: 7436-7442.
• 6 Moschen AR, Gerner R, Schroll A, Fritz T, Kaser A, Tilg H. A key role for Pre-B cell colony-enhancing factor in experimental hepatitis. Hepatology 2011; 54: 675-686.
• 7 Weber C, Noels H. Atherosclerosis: current pathogenesis and therapeutic options. Nat Med 2011; 17: 1410-1422.
• 8 Koenen RR, Weber C. Chemokines: established and novel targets in atherosclerosis. EMBO Mol Med 2011; 3: 713-725.
• 9 Soehnlein O. Multiple roles for neutrophils in atherosclerosis. Circ Res 2012; 110: 875-888.
• 10 Montecucco F, Di Marzo V, da Silva RF. et al. The activation of the cannabinoid receptor type 2 reduces neutrophilic protease-mediated vulnerability in atherosclerotic plaques. Eur Heart J 2012; 33: 846-856.
• 11 Montecucco F, Lenglet S, Gayet-Ageron A. et al. Systemic and intraplaque mediators of inflammation are increased in patients symptomatic for ischaemic stroke. Stroke 2010; 41: 1394-1404.
• 12 Copin JC, da Silva RF, Fraga-Silva RA. et al. Treatment with Evasin-3 reduces atherosclerotic vulnerability for ischaemic stroke, but not brain injury in mice. Cereb Blood Flow Metab 2013; 33: 490-498.
• 13 Koenen RR, Weber C. Manipulating the chemokine system: therapeutic perspectives for atherosclerosis. Curr Opin Investig Drugs 2010; 11: 265-272.
• 14 Koenen RR, Weber C. Therapeutic targeting of chemokine interactions in atherosclerosis. Nat Rev Drug Discov 2010; 9: 141-153.
• 15 Dahl TB, Yndestad A, Skjelland M. et al. Increased expression of visfatin in mac-rophages of human unstable carotid and coronary atherosclerosis: possible role in inflammation and plaque destabilisation. Circulation 2007; 115: 972-980.
• 16 Cheng C, Tempel D, van Haperen R. et al. Atherosclerotic lesion size and vulnerability are determined by patterns of fluid shear stress. Circulation 2006; 113: 2744-2753.
• 17 Lenglet S, Thomas A, Soehnlein O. et al. Fatty acid amide hydrolase deficiency enhances intraplaque neutrophil recruitment in atherosclerotic mice. Arte-rioscler Thromb Vasc Biol 2013; 33: 215-223.
• 18 van Leeuwen M, Gijbels MJ, Duijvestijn A. et al. Accumulation of myeloperox-idase-positive neutrophils in atherosclerotic lesions in LDLR-/- mice. Arte- rioscler Thromb Vasc Biol 2008; 28: 84-89.
• 19 Polykratis A, van Loo G, Xanthoulea S, Hellmich M, Pasparakis M. Conditional targeting of tumor necrosis factor receptor-associated factor 6 reveals opposing functions of Toll-like receptor signalling in endothelial and myeloid cells in a mouse model of atherosclerosis. Circulation 2012; 126: 1739-1751.
• 20 Skehan P, Storeng R, Scudiero D. et al. New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 1990; 82: 1107-1112.
• 21 Graeff R, Lee HC. A novel cycling assay for cellular cADP-ribose with nanomo- lar sensitivity. Biochem J 2002; 361: 379-384.
• 22 Skokowa J, Lan D, Thakur BK. et al. NAMPT is essential for the G-CSF-induced myeloid differentiation via a NAD(+)-sirtuin-1-dependent pathway. Nat Med 2009; 15: 151-158.
• 23 Quercioli A, Mach F, Bertolotto M. et al. Receptor activator of NF- kB ligand (RANKL) increases the release of neutrophil products associated with coronary vulnerability. Thromb Haemost 2012; 107: 124-139.
• 24 Shattuck RL, Wood LD, Jaffe GJ, Richmond A. MGSA/GRO transcription is differentially regulated in normal retinal pigment epithelial and melanoma cells. Mol Cell Biol 1994; 14: 791-802.
• 25 Wood LD, Farmer AA, Richmond A. HMGI(Y) and Sp1 in addition to NF- kappa B regulate transcription of the MGSA/GRO alpha gene. Nucleic Acids Res 1995; 23: 4210-4219.
• 26 Ohtsuka T, Kubota A, Hirano T. et al. Glucocorticoid-mediated gene suppression of rat cytokine-induced neutrophil chemoattractant CINC/gro, a member of the interleukin-8 family, through impairment of NF-kappa B activation. J Biol Chem 1996; 271: 1651-1659.
• 27 Pierce JW, Schoenleber R, Jesmok G. et al. Novel inhibitors of cytokine-induced IkappaBalpha phosphorylation and endothelial cell adhesion molecule expression show anti-inflammatory effects in vivo. J Biol Chem 1997; 272: 21096-21103.
• 28 Olsen LS, Hjarnaa PJ, Latini S. et al. Anticancer agent CHS 828 suppresses nuclear factor-kappa B activity in cancer cells through downregulation of IKK activity. Int J Cancer 2004; 111: 198-205.
• 29 Hassan SB, Lövborg H, Lindhagen E. et al. CHS 828 kill tumour cells by inhibiting the nuclear factor-kappaB translocation but unlikely through down-regulation of proteasome. Anticancer Res 2006; 26: 4431-4436.
• 30 Esposito E, Impellizzeri D, Mazzon E. et al. The NAMPT inhibitor FK866 reverts the damage in spinal cord injury. J Neuroinflammation 2012; 9: 66.
• 31 Soehnlein O, Drechsler M, Döring Y. et al. Distinct functions of chemokine receptor axes in the atherogenic mobilisation and recruitment of classical mono-cytes. EMBO Mol Med 2013; 5: 471-481.
• 32 Boisvert WA, Rose DM, Johnson KA. et al. Up-regulated expression of the CXCR2 ligand KC/GRO-alpha in atherosclerotic lesions plays a central role in macrophage accumulation and lesion progression. Am J Pathol 2006; 168: 1385-1395.
• 33 Montecucco F, Lenglet S, Braunersreuther V. et al. Single administration of the CXC chemokine-binding protein Evasin-3 during ischaemia prevents myo-cardial reperfusion injury in mice. Arterioscler Thromb Vasc Biol 2010; 30: 1371-1377.
• 34 Wan W, Murphy PM. Regulation of atherogenesis by chemokines and chemo-kine receptors. Arch Immunol Ther Exp 2013; 61: 1-14.
• 35 Drechsler M, Megens RT, van Zandvoort M. et al. Hyperlipidemia-triggered neutrophilia promotes early atherosclerosis. Circulation 2010; 122: 1837-1845.
• 36 Shimomura I, Shimano H, Korn BS. et al. Nuclear sterol regulatory element-binding proteins activate genes responsible for the entire program of unsatu-rated fatty acid biosynthesis in transgenic mouse liver. J Biol Chem 1998; 273: 35299-35306.