Kynurenic acid and its derivatives are able to modulate the adhesion and locomotion of brain endothelial cells

Abstract

The neuroprotective actions of kynurenic acid (KYNA) and its derivatives in several neurodegenerative disorders [characterized by damage to the cerebral endothelium and to the blood–brain barrier (BBB)] are well established. Cell–extracellular matrix (ECM) adhesion is supposedly involved in recovery of impaired cerebral endothelium integrity (endothelial repair). The present work aimed to investigate the effects of KYNA and its synthetic derivatives on cellular behaviour (e.g. adhesion and locomotion) and on morphology of the GP8 rat brain endothelial cell line, modeling the BBB endothelium. The effects of KYNA and its derivatives on cell adhesion were measured using an impedance-based technique, the xCELLigence SP system. Holographic microscopy (Holomonitor M4) was used to analyse both chemokinetic responses and morphometry. The GP8 cells proved to be a suitable model cell line for investigating cell adhesion and the locomotion modulator effects of kynurenines. KYNA enhanced cell adhesion and spreading, and also decreased the migration/motility of GP8 cells at physiological concentrations (10−9 and 10−7 mol/L). The derivatives containing an amide side-chain at the C2 position (KYNA-A1 and A2) had lower adhesion inducer effects compared to KYNA. All synthetic analogues (except KYNA-A5) had a time-dependent inhibitory effect on GP8 cell adhesion at a supraphysiological concentration (10−3 mol/L). The immobilization promoting effect of KYNA and the adhesion inducer activity of its derivatives indicate that these compounds could contribute to maintaining or restoring the protective function of brain endothelium; they also suggest that cell–ECM adhesion and related cell responses (e.g. migration/motility) could be potential new targets of KYNA.

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References

  1. Andras IE, Deli MA, Veszelka S, Hayashi K, Hennig B, Toborek M (2007) The NMDA and AMPA/KA receptors are involved in glutamate-induced alterations of occludin expression and phosphorylation in brain endothelial cells. J Cereb Blood Flow Metab 27:1431–1443. https://doi.org/10.1038/sj.jcbfm.9600445

    Article  PubMed  CAS  Google Scholar 

  2. Antal O, Hackler JL, Shen J, Mán I, Hideghéty K, Kitajka K, Puskás LG (2014) Combination of unsaturated fatty acids and ionizing radiation on human glioma cells: cellular, biochemical and gene expression analysis. Lipids Health Dis 13:142–157. https://doi.org/10.1186/1476-511x-13-142

    Article  PubMed  PubMed Central  Google Scholar 

  3. Backer H et al (2017) Impedimetric analysis of the effect of decellularized porcine heart scaffold on human fibrosarcoma, endothelial, and cardiomyocyte cell lines. Med Sci Monit 23:2232–2240

    Article  PubMed  PubMed Central  Google Scholar 

  4. Baeten KM, Akassoglou K (2011) Extracellular matrix and matrix receptors in blood–brain barrier formation and stroke. Dev Neurobiol 71:1018–1039. https://doi.org/10.1002/dneu.20954

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  5. Baran H, Amann G, Lubec B, Lubec G (1997) Kynurenic acid and kynurenine aminotransferase in heart. Pediatr Res 41:404–410. https://doi.org/10.1203/00006450-199703000-00017

    Article  PubMed  CAS  Google Scholar 

  6. Barth MC et al (2009) Kynurenic acid triggers firm arrest of leukocytes to vascular endothelium under flow conditions. J Biol Chem 284:19189–19195. https://doi.org/10.1074/jbc.M109.024042

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  7. Beal MF, Matson WR, Swartz KJ, Gamache PH, Bird ED (1990) Kynurenine pathway measurements in Huntington’s disease striatum: evidence for reduced formation of kynurenic acid. J Neurochem 55:1327–1339. https://doi.org/10.1111/j.1471-4159.1990.tb03143.x

    Article  PubMed  CAS  Google Scholar 

  8. Besler C, Doerries C, Giannotti G, Luscher TF, Landmesser U (2008) Pharmacological approaches to improve endothelial repair mechanisms. Expert Rev Cardiovasc Ther 6:1071–1082. https://doi.org/10.1586/14779072.6.8.1071

    Article  PubMed  CAS  Google Scholar 

  9. Cosi C et al (2011) G-protein coupled receptor 35 (GPR35) activation and inflammatory pain: studies on the antinociceptive effects of kynurenic acid and zaprinast. Neuropharmacology 60:1227–1231. https://doi.org/10.1016/j.neuropharm.2010.11.014S0028-3908(10)00313-8

    Article  PubMed  CAS  Google Scholar 

  10. Culot M et al (2008) An in vitro blood–brain barrier model for high throughput (HTS) toxicological screening. Toxicol In Vitro 22:799–811. https://doi.org/10.1016/j.tiv.2007.12.016

    Article  PubMed  CAS  Google Scholar 

  11. Dantzer R, O’Connor JC, Freund GG, Johnson RW, Kelley KW (2008) From inflammation to sickness and depression: when the immune system subjugates the brain. Nat Rev Neurosci 9:46–56. https://doi.org/10.1038/nrn2297

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  12. Deanfield JE, Halcox JP, Rabelink TJ (2007) Endothelial function and dysfunction: testing and clinical relevance. Circulation 115:1285–1295. https://doi.org/10.1161/circulationaha.106.652859

    PubMed  Article  Google Scholar 

  13. Dehouck MP, Meresse S, Delorme P, Fruchart JC, Cecchelli R (1990) An easier, reproducible, and mass-production method to study the blood–brain barrier in vitro. J Neurochem 54:1798–1801. https://doi.org/10.1111/j.1471-4159.1990.tb01236.x

    Article  PubMed  CAS  Google Scholar 

  14. del Zoppo GJ, Milner R (2006) Integrin–matrix interactions in the cerebral microvasculature. Arterioscler Thromb Vasc Biol 26:1966–1975. https://doi.org/10.1161/01.ATV.0000232525.65682.a2

    Article  PubMed  CAS  Google Scholar 

  15. Demeter I, Nagy K, Gellert L, Vecsei L, Fulop F, Toldi J (2012) A novel kynurenic acid analog (SZR104) inhibits pentylenetetrazole-induced epileptiform seizures. An electrophysiological study: special issue related to kynurenine. J Neural Transm 119:151–154. https://doi.org/10.1007/s00702-011-0755-x

    Article  PubMed  CAS  Google Scholar 

  16. Dezsi L, Tuka B, Martos D, Vecsei L (2015) Alzheimer’s disease, astrocytes and kynurenines. Curr Alzheimer Res 12:462–480

    Article  PubMed  CAS  Google Scholar 

  17. Fülöp F, Szatmári I, Toldi J, Vécsei L (2012) Modifications on the carboxylic function of kynurenic acid. J Neural Transm 119:109–114. https://doi.org/10.1007/s00702-011-0721-7

    Article  PubMed  CAS  Google Scholar 

  18. Greenwood J, Pryce G, Devine L, Male DK, dos Santos WL, Calder VL, Adamson P (1996) SV40 large T immortalised cell lines of the rat blood–brain and blood–retinal barriers retain their phenotypic and immunological characteristics. J Neuroimmunol 71:51–63. https://doi.org/10.1016/S0165-5728(96)00130-0

    Article  PubMed  CAS  Google Scholar 

  19. Han Q, Cai T, Tagle DA, Li J (2010) Structure, expression, and function of kynurenine aminotransferases in human and rodent brains. Cell Mol Life Sci 67:353–368. https://doi.org/10.1007/s00018-009-0166-4

    Article  PubMed  CAS  Google Scholar 

  20. Hilmas C, Pereira EF, Alkondon M, Rassoulpour A, Schwarcz R, Albuquerque EX (2001) The brain metabolite kynurenic acid inhibits alpha7 nicotinic receptor activity and increases non-alpha7 nicotinic receptor expression: physiopathological implications. J Neurosci 21:7463–7473

    Article  PubMed  CAS  Google Scholar 

  21. Kandanearatchi A, Brew BJ (2012) The kynurenine pathway and quinolinic acid: pivotal roles in HIV associated neurocognitive disorders. FEBS J 279:1366–1374. https://doi.org/10.1111/j.1742-4658.2012.08500.x

    Article  PubMed  CAS  Google Scholar 

  22. Kelleher RJ, Soiza RL (2013) Evidence of endothelial dysfunction in the development of Alzheimer’s disease: is Alzheimer’s a vascular disorder? Am J Cardiovasc Dis 3:197–226

    PubMed  PubMed Central  Google Scholar 

  23. Kis B et al (1999) Vasoactive substances produced by cultured rat brain endothelial cells. Eur J Pharmacol 368:35–42. https://doi.org/10.1016/S0014-2999(99)00024-2

    Article  PubMed  CAS  Google Scholar 

  24. Krizbai IA, Deli MA, Pestenacz A, Siklos L, Szabo CA, Andras I, Joo F (1998) Expression of glutamate receptors on cultured cerebral endothelial cells. J Neurosci Res 54:814–819. https://doi.org/10.1002/(sici)1097-4547(19981215)54:6<814::aid-jnr9>3.0.co;2-3

  25. Lajkó E, Bányai P, Zámbó Z, Kursinszki L, Szőke É, Kőhidai L (2015) Targeted tumor therapy by Rubia tinctorum L.: analytical characterization of hydroxyanthraquinones and investigation of their selective cytotoxic, adhesion and migration modulator effects on melanoma cell line (A2058 and HT168-M1). Cancer Cell. https://doi.org/10.1186/s12935-015-0271-4

    Article  Google Scholar 

  26. Leurs U et al (2012) GnRH-III based multifunctional drug delivery systems containing daunorubicin and methotrexate. Eur J Med Chem 52:173–183. https://doi.org/10.1016/j.ejmech.2012.03.016

    Article  PubMed  CAS  Google Scholar 

  27. Lyros E, Bakogiannis C, Liu Y, Fassbender K (2014) Molecular links between endothelial dysfunction and neurodegeneration in Alzheimer’s disease. Curr Alzheimer Res 11:18–26

    Article  PubMed  CAS  Google Scholar 

  28. Mandi Y, Vecsei L (2012) The kynurenine system and immunoregulation. J Neural Transm 119:197–209. https://doi.org/10.1007/s00702-011-0681-y

    Article  PubMed  CAS  Google Scholar 

  29. Persson J, Mölder A, Pettersson SG, Alm K (2010) Cell motility studies using digital holographic microscopy. In: Vilas M, Díaz J (eds) Microscopy: science, technology, applications and education, vol 4. Formatex Research Center, Badajoz, Spain, pp 1063–1072

    Google Scholar 

  30. Moroni F, Cozzi A, Sili M, Mannaioni G (2012) Kynurenic acid: a metabolite with multiple actions and multiple targets in brain and periphery. J Neural Transm 119:133–139. https://doi.org/10.1007/s00702-011-0763-x

    Article  PubMed  CAS  Google Scholar 

  31. Nagy K et al (2011) Synthesis and biological effects of some kynurenic acid analogs. Bioorg Med Chem 19:7590–7596. https://doi.org/10.1016/j.bmc.2011.10.029S0968-0896(11)00837-6

    Article  PubMed  CAS  Google Scholar 

  32. Ogawa T, Matson WR, Beal MF, Myers RH, Bird ED, Milbury P, Saso S (1992) Kynurenine pathway abnormalities in Parkinson’s disease. Neurology 42:1702–1706

    Article  PubMed  CAS  Google Scholar 

  33. Oláh G et al (2013) Unexpected effects of peripherally administered kynurenic acid on cortical spreading depression and related blood–brain barrier permeability. Drug Des Dev Ther 7:981–987. https://doi.org/10.2147/dddt.s44496

    Article  Google Scholar 

  34. Owe-Young R et al (2008) Kynurenine pathway metabolism in human blood–brain-barrier cells: implications for immune tolerance and neurotoxicity. J Neurochem 105:1346–1357. https://doi.org/10.1111/j.1471-4159.2008.05241.xJNC5241

    Article  PubMed  CAS  Google Scholar 

  35. Pawlak K, Kowalewska A, Mysliwiec M, Pawlak D (2009) Kynurenine and its metabolites–kynurenic acid and anthranilic acid are associated with soluble endothelial adhesion molecules and oxidative status in patients with chronic kidney disease. Am J Med Sci 338:293–300. https://doi.org/10.1097/MAJ.0b013e3181aa30e6

    Article  PubMed  Google Scholar 

  36. Pawlak K, Mysliwiec M, Pawlak D (2010) Kynurenine pathway—a new link between endothelial dysfunction and carotid atherosclerosis in chronic kidney disease patients. Adv Med Sci 55:196–203. https://doi.org/10.2478/v10039-010-0015-6

    Article  PubMed  CAS  Google Scholar 

  37. Perkins MN, Stone TW (1982) An iontophoretic investigation of the actions of convulsant kynurenines and their interaction with the endogenous excitant quinolinic acid. Brain Res 247:184–187

    Article  PubMed  CAS  Google Scholar 

  38. Persidsky Y, Ramirez SH, Haorah J, Kanmogne GD (2006) Blood–brain barrier: structural components and function under physiologic and pathologic conditions. J Neuroimmune Pharmacol 1:223–236. https://doi.org/10.1007/s11481-006-9025-3

    Article  PubMed  Google Scholar 

  39. Peter B et al (2015) Incubator proof miniaturized Holomonitor to in situ monitor cancer cells exposed to green tea polyphenol and preosteoblast cells adhering on nanostructured titanate surfaces: validity of the measured parameters and their corrections. J Biomed Opt 20:067002. https://doi.org/10.1117/1.jbo.20.6.067002

    Article  PubMed  Google Scholar 

  40. Phase Holographic Imaging PHI AB (2015) HoloMonitor application note on Label-free Cell Motility. Phase Holographic Imaging PHI AB. http://www.phiab.se/reports/2014/CellMotilityAppNotePHI-140919.pdf. Accessed 29 Sept 2017

  41. Prescott C, Weeks AM, Staley KJ, Partin KM (2006) Kynurenic acid has a dual action on AMPA receptor responses. Neurosci Lett 402:108–112. https://doi.org/10.1016/j.neulet.2006.03.051

    Article  PubMed  CAS  Google Scholar 

  42. Regina A, Romero IA, Greenwood J, Adamson P, Bourre JM, Couraud PO, Roux F (1999) Dexamethasone regulation of P-glycoprotein activity in an immortalized rat brain endothelial cell line, GPNT. J Neurochem 73:1954–1963. https://doi.org/10.1046/j.1471-4159.1999.01954.x

    PubMed  CAS  Article  Google Scholar 

  43. Roux F, Couraud PO (2005) Rat brain endothelial cell lines for the study of blood–brain barrier permeability and transport functions. Cell Mol Neurobiol 25:41–58. https://doi.org/10.1007/s10571-004-1376-9

    Article  PubMed  Google Scholar 

  44. Rozsa E, Robotka H, Vecsei L, Toldi J (2008) The Janus-face kynurenic acid. J Neural Transm 115:1087–1091. https://doi.org/10.1007/s00702-008-0052-5

    Article  PubMed  CAS  Google Scholar 

  45. Silva-Adaya D et al (2011) Protective effect of l-kynurenine and probenecid on 6-hydroxydopamine-induced striatal toxicity in rats: implications of modulating kynurenate as a protective strategy. Neurotoxicol Teratol 33:303–312. https://doi.org/10.1016/j.ntt.2010.10.002S0892-0362(10)00171-6

    Article  PubMed  CAS  Google Scholar 

  46. Stazka J, Luchowski P, Wielosz M, Kleinrok Z, Urbanska EM (2002) Endothelium-dependent production and liberation of kynurenic acid by rat aortic rings exposed to l-kynurenine. Eur J Pharmacol 448:133–137. https://doi.org/10.1016/S0014-2999(02)01943-X

    Article  PubMed  CAS  Google Scholar 

  47. Stazka J, Luchowski P, Urbanska EM (2005) Homocysteine, a risk factor for atherosclerosis, biphasically changes the endothelial production of kynurenic acid. Eur J Pharmacol 517:217–223. https://doi.org/10.1016/j.ejphar.2005.04.048

    Article  PubMed  CAS  Google Scholar 

  48. Tang W, Song H, Cai W, Shen X (2015) Real time monitoring of inhibition of adipogenesis and angiogenesis by (−)-epigallocatechin-3-gallate in 3T3-L1 adipocytes and human umbilical vein endothelial cells. Nutrients 7:8871–8886. https://doi.org/10.3390/nu7105437

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  49. Tiszlavicz Z, Nemeth B, Fulop F, Vecsei L, Tapai K, Ocsovszky I, Mandi Y (2011) Different inhibitory effects of kynurenic acid and a novel kynurenic acid analogue on tumour necrosis factor-alpha (TNF-alpha) production by mononuclear cells, HMGB1 production by monocytes and HNP1-3 secretion by neutrophils. Naunyn Schmiedebergs Arch Pharmacol 383:447–455. https://doi.org/10.1007/s00210-011-0605-2

    Article  PubMed  CAS  Google Scholar 

  50. Tóth AE et al (2014) Edaravone protects against methylglyoxal-induced barrier damage in human brain endothelial cells. PLoS One 9:e100152. https://doi.org/10.1371/journal.pone.0100152

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  51. Urcan E, Haertel U, Styllou M, Hickel R, Scherthan H, Reichl FX (2010) Real-time xCELLigence impedance analysis of the cytotoxicity of dental composite components on human gingival fibroblasts. Dent Mater 26:51–58. https://doi.org/10.1016/j.dental.2009.08.007

    Article  PubMed  CAS  Google Scholar 

  52. Varga G et al (2010) N-Methyl-d-aspartate receptor antagonism decreases motility and inflammatory activation in the early phase of acute experimental colitis in the rat. Neurogastroenterol Motil 22(217–225):e268. https://doi.org/10.1111/j.1365-2982.2009.01390.xNMO1390

    Article  Google Scholar 

  53. Vecsei L, Szalardy L, Fulop F, Toldi J (2013) Kynurenines in the CNS: recent advances and new questions. Nat Rev Drug Discov 12:64–82. https://doi.org/10.1038/nrd3793nrd3793

    Article  PubMed  CAS  Google Scholar 

  54. Wejksza K et al (2004) Kynurenic acid production in cultured bovine aortic endothelial cells. Homocysteine is a potent inhibitor. Naunyn Schmiedebergs Arch Pharmacol 369:300–304. https://doi.org/10.1007/s00210-004-0872-2

    Article  PubMed  CAS  Google Scholar 

  55. Wejksza K, Rzeski W, Turski WA (2009) Kynurenic acid protects against the homocysteine-induced impairment of endothelial cells. Pharmacol Rep 61:751–756

    Article  PubMed  CAS  Google Scholar 

  56. Weksler BB et al (2005) Blood–brain barrier-specific properties of a human adult brain endothelial cell line. FASEB J 19:1872–1874. https://doi.org/10.1096/fj.04-3458fje

    Article  PubMed  CAS  Google Scholar 

  57. Wilhelm I, Fazakas C, Krizbai IA (2011) In vitro models of the blood–brain barrier. Acta Neurobiol Exp (Wars) 71:113–128

    Google Scholar 

  58. Wu HQ, Ungerstedt U, Schwarcz R (1995) l-alpha-aminoadipic acid as a regulator of kynurenic acid production in the hippocampus: a microdialysis study in freely moving rats. Eur J Pharmacol 281:55–61

    Article  PubMed  CAS  Google Scholar 

  59. Zadori D et al (2011) Neuroprotective effects of a novel kynurenic acid analogue in a transgenic mouse model of Huntington’s disease. J Neural Transm 118:865–875. https://doi.org/10.1007/s00702-010-0573-6

    Article  PubMed  CAS  Google Scholar 

  60. Zampetaki A, Kirton JP, Xu Q (2008) Vascular repair by endothelial progenitor cells. Cardiovasc Res 78:413–421. https://doi.org/10.1093/cvr/cvn081

    Article  PubMed  CAS  Google Scholar 

  61. Zwilling D et al (2011) Kynurenine 3-monooxygenase inhibition in blood ameliorates neurodegeneration. Cell 145:863–874. https://doi.org/10.1016/j.cell.2011.05.020S0092-8674(11)00581-2

    Article  PubMed  PubMed Central  CAS  Google Scholar 

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Acknowledgements

Authors express their gratitude to Auro-Science Consulting Kft for their expert technical assistance, and to Mr. P. Samotik and Mr. Z. Ambrus for proofreading and technical language support.

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Correspondence to László Kőhidai.

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This work was supported by the following grants: GINOP-2.3.2-15-2016-00034, MTA-SZTE Neuroscience Research Group. The funding sources were in no way involved in the conduct of the research, nor in the analysis or interpretation of data, nor in preparation of the article.

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Kálmán Magyar : Deceased.

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Lajkó, E., Tuka, B., Fülöp, F. et al. Kynurenic acid and its derivatives are able to modulate the adhesion and locomotion of brain endothelial cells. J Neural Transm 125, 899–912 (2018). https://doi.org/10.1007/s00702-018-1839-7

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Keywords

  • Endothelial repair
  • Kynurenic acid
  • Holographic microscopy
  • Impedimetry
  • Synthetic kynurenines
  • Brain endothelial cell
  • Kynurenic acid (PubChem CID: 3845)
  • l-Kynurenine (Pubchem CID: 161166)