Urinary neopterin and nitric oxide metabolites as markers of interferon β-1a activity in primary progressive multiple sclerosis

Abstract

Background: Interferon beta has not been demonstrated to be effective in exploratory phase 2 clinical trials in primary progressive multiple sclerosis. However, using more sensitive indicators of a treatment response, such as biomarkers, might help to identify sub-groups of patients who may benefit from therapy.

Objective: To assess the utility of measuring urinary neopterin and nitric oxide metabolite excretion for monitoring interferon β-1a (IFNβ-1a) treatment in patients with primary progressive multiple sclerosis.

Methods: Fifty patients from a phase II trial of IFNβ-1a (Placebo n = 20; Avonex^® 1 × 30 μg/week (IFN-30), n = 15; Avonex^® 1 × 60 μg/week (IFN-60), n = 15), were enrolled. Patients were assessed using the Expanded Disability Status Scale. Urine samples were collected on each visit, 3 months apart, for a period of 24 months. Nitric oxide metabolites, nitrite/nitrate (NOx), were measured by colorimetric assay and neopterin and creatinine (Cr) were assayed using a high-performance liquid chromatography technique. NOx/creatinine ratio (NOxCR) and urinary neopterin/creatinine ratio (UNCR) quotients were calculated.

Results: There was no significant difference between pre-dose, baseline levels of UNCR or NOxCR between the study groups. On the intention-to-treat analysis, there was a significant difference in UNCR levels between the placebo compared with IFN-30 (p = 0.03) or IFN-60 (p = 0.002) groups. The IFN-30 and IFN-60 groups did not differ. Within IFNβ-1a-treated patients with primary progressive multiple sclerosis, median UNCR values were significantly higher in clinically stable (no Expanded Disability Status Scale change) compared with progressive patients (p = 0.002). IFNβ-1a treatment did not significantly influence NOx excretion in patients with primary progressive multiple sclerosis.

Conclusions: Urinary neopterin is a potential biomarker to monitor the in vivo effects of IFNβ-1a in primary progressive multiple sclerosis and other multiple sclerosis sub-types.

Keywords

beta-interferon biomarkers multiple sclerosis neopterin nitric oxide progressive

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References

Leary SM , Miller DH , Stevenson VL , Brex PA , Chard DT and Thompson AJ Interferon beta-1a in primary progressive MS: an exploratory, randomized, controlled trial. Neurology 2003; 60: 44-51.

Montalban X. Overview of European pilot study of interferon beta-Ib in primary progressive multiple sclerosis. Mult Scler 2004; 10(Suppl 1): S62.

Blackhall F. , Ranson M. and Thatcher N.

Where next for gefitinib in patients with lung cancer?

Lancet Oncol 2006; 7: 499-507.

Boylan MT , Crockard AD , Duddy ME , Armstrong MA , McMillan SA and Hawkins SA Interferon-beta1a administration results in a transient increase of serum amyloid A protein and C-reactive protein: comparison with other markers of inflammation. Immunol Lett 2001; 75: 191-197.

Satoh J. , Nanri Y. , Tabunoki H. and Yamamura T. Microarray analysis identifies a set of CXCR3 and CCR2 ligand chemokines as early IFNbeta-responsive genes in peripheral blood lymphocytes in vitro: an implication for IFNbeta-related adverse effects in multiple sclerosis. BMC Neurol 2006; 6: 18.

Williams GJ and Witt PL Comparative study of the pharmacodynamic and pharmacologic effects of Betaseron and AVONEX. J Interferon Cytokine Res 1998 ; 18: 967-975.

Bagnato F. , Pozzilli C. , Scagnolari C. , et al. A one-year study on the pharmacodynamic profile of interferon-beta1a in MS. Neurology 2002; 58: 1409-1411.

Werner ER , Werner-Felmayer G. , Fuchs D. , et al. Biochemistry and function of pteridine synthesis in human and murine macrophages. Pathobiology 1991; 59: 276-279.

Werner-Felmayer G. , Werner ER , Fuchs D. , et al. Pteridine biosynthesis in human endothelial cells. Impact on nitric oxide-mediated formation of cyclic GMP. J Biol Chem 1993; 268: 1842-1846.

10.

Andert SE , Griesmacher A. , Zuckermann A. and Muller MM Neopterin release from human endothelial cells is triggered by interferon-gamma . Clin Exp Immunol 1992; 88: 555-558.

11.

Rejdak K. , Empl M. , Giffin NJ , et al. Increased urinary excretion of nitric oxide metabolites in longitudinally monitored migraine patients. Eur J Neurol 2006; 13: 1346-1351.

12.

Hua LL , Liu JS , Brosnan CF and Lee SC Selective inhibition of human glial inducible nitric oxide synthase by interferon-beta: implications for multiple sclerosis. Ann Neurol 1998; 43: 384-387.

13.

Hua LL , Kim MO , Brosnan CF and Lee SC Modulation of astrocyte inducible nitric oxide synthase and cytokine expression by interferon beta is associated with induction and inhibition of interferon gamma-activated sequence binding activity. J Neurochem 2002; 83: 1120-1128.

14.

Lieb K. , Engels S. and Fiebich BL Inhibition of LPSinduced iNOS and NO synthesis in primary rat microglial cells . Neurochem Int 2003; 42: 131-137.

15.

Rejdak K. , Petzold A. , Sharpe MA , et al. Serum and urine nitrate and nitrite are not reliable indicators of intrathecal nitric oxide production in acute brain injury. J Neurol Sci 2003; 208: 1-7.

16.

Miranda KM , Espey MG and Wink DA A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide 2001; 5: 62-71.

17.

Giovannoni G. , Lai M. , Kidd D. , et al. Daily urinary neopterin excretion as an immunological marker of disease activity in multiple sclerosis. Brain 1997; 120: 1-13.

18.

Littell RC , Henry PR and Ammerman CB Statistical analysis of repeated measures data using SAS procedures. J Anim Sci 1998; 76: 1216-1231.

19.

Cook SD , Quinless JR , Jotkowitz A. and Beaton P. and the Neutralizing Antibody Study Group. Serum IFN neutralizing antibodies and neopterin levels in a cross-section of MS patients. Neurology 2001; 57: 1080-1084.

20.

Millonig A. , Dressel A. , Bahner D. , et al. MxA protein - an interferon beta biomarker in primary progressive multiple sclerosis patients. Eur J Neurol 2008; 15: 822-826.

21.

Giovannoni G. and Thompson EJ Urinary markers of disease activity in multiple sclerosis. Mult Scler 1998; 4: 247-253.

22.

Rejdak K. , Eikelenboom MJ , Petzold A. , et al. CSF nitric oxide metabolites are associated with activity and progression of multiple sclerosis. Neurology 2004; 63: 1439-1445.

23.

Giovannoni G. , Heales SJ , Silver NC , et al. Raised serum nitrate and nitrite levels in patients with multiple sclerosis. J Neurol Sci 1997; 145: 77-81.

24.

Giovannoni G. , Silver NC , O’Riordan J. , et al. Increased urinary nitric oxide metabolites in patients with multiple sclerosis correlates with early and relapsing disease. Mult Scler 1999 ; 5: 335-341.

25.

Kurioka S. , Koshimura K. , Sugitani M. , Murakami Y. , Nishiki M. and Kato Y. Analysis of urinary nitric oxide metabolites in healthy subjects. Endocr J 1999; 46: 421-428.

26.

Kwon NS , Nathan CF and Stuehr DJ Reduced biopterin as a cofactor in the generation of nitrogen oxides by murine macrophages. J Biol Chem 1989; 264: 20496-20501.

27.

Lopez-Collazo E. , Hortelano S. , Rojas A. and Bosca L. Triggering of peritoneal macrophages with IFN-alpha/ beta attenuates the expression of inducible nitric oxide synthase through a decrease in NF-kappaB activation. J Immunol 1998; 160: 2889-2895.