Horm Metab Res 2009; 41(6): 465-470
DOI: 10.1055/s-0029-1214413
Humans, Clinical

© Georg Thieme Verlag KG Stuttgart · New York

Proteomics in Autoimmune Thyroid Eye Disease

R. Okrojek 1 , F. H. Grus 2 , N. Matheis 1 , G. J. Kahaly 1
  • 1Thyroid Research Laboratory, Department of Medicine I, Gutenberg University Hospital, Mainz, Germany
  • 2Experimental Ophthalmology, Department of Ophthalmology, Gutenberg University Hospital, Mainz, Germany
Further Information

Publication History

received 18.12.2008

accepted 11.02.2009

Publication Date:
16 April 2009 (online)

Abstract

Ocular and systemic autoimmune diseases impair the proteome patterns of tear fluid. To learn more about the complex pathological processes in autoimmune thyroid eye disease (TED) it is essential to get detailed information on these proteins. Therefore, the purpose of this prospective and controlled study was to detect and evaluate possible changes in the proteomic patterns in tear fluid of patients with TED. Tear samples from TED patients with various disease severity and activity, and healthy controls were analyzed with the SELDI-TOF-MS-technology using arrays with different chromatographic surfaces (CM10 cation exchange, H50 reversed-phase). Data were analyzed by multivariate statistical techniques and artificial neural networks. The discriminate analysis revealed significant changes (p<0.05) in the protein profiles of TED patients compared to controls. We obtained a set of protein biomarkers that allowed us to clearly discriminate between patients and controls with a very high sensitivity and specificity (ROC curve, r=0.99). All possible biomarkers found in this study had a molecular weight between 3000 and 20 000 Da. The majority of the proteins was downregulated in the patient group, with only few proteins overexpressed in comparison to healthy controls. The SELDI-TOF-MS is an accurate method for proteome analysis in tear fluid of TED patients. These proteins may serve as biomarkers for diagnosis and follow-up during treatment.

References

  • 1 Bahn RS, Heufelder AE. Pathogenesis of Graves’ ophthalmopathy.  N Engl J Med. 1993;  329 1468-1475
  • 2 Prabhakar BS, Bahn RS, Smith TJ. Current perspective on the pathogenesis of Graves’ disease and ophthalmopathy.  Endocr Rev. 2003;  24 802-835
  • 3 Bahn RS, Dutton CM, Natt N, Joba W, Spitzweg C, Heufelder AE. Thyrotropin receptor expression in Graves' orbital adipose/connective tissues: potential autoantigen in Graves' ophthalmopathy.  J Clin Endocrinol Metab. 1998;  83 998-1002
  • 4 Eckstein AK, Finkenrath A, Heiligenhaus A, Renzing-Kohler K, Esser J, Kruger C, Quadbeck B, Steuhl KP, Gieseler RK. Dry eye syndrome in thyroid-associated ophthalmopathy: lacrimal expression of TSH receptor suggests involvement of TSHR-specific autoantibodies.  Acta Ophthalmol Scand. 2004;  82 291-297
  • 5 Giovanella L, Ceriani L, Ghelfo A. Second-generation thyrotropin receptor antibodies assay and quantitative thyroid scintigraphy in autoimmune hyperthyroidism.  Horm Metab Res. 2008;  40 484-486
  • 6 Gunji K, Kubota S, Stolarski C, Wengrowicz S, Kennerdell JS, Wall JR. A 63 kDa skeletal muscle protein associated with eye muscle inflammation in Graves' disease is identified as the calcium binding protein calsequestrin.  Autoimmunity. 1999;  29 1-9
  • 7 Kubota S, Gunji K, Ackrell BA, Cochran B, Stolarski C, Wengrowicz S, Kennerdell JS, Hiromatsu Y, Wall J. The 64-kilodalton eye muscle protein is the flavoprotein subunit of mitochondrial succinate dehydrogenase: the corresponding serum antibodies are good markers of an immune-mediated damage to the eye muscle in patients with Graves' hyperthyroidism.  J Clin Endocrinol Metab. 1998;  83 443-447
  • 8 Kahaly GJ, Bang H, Berg W, Dittmar M. Alpha-fodrin as a putative autoantigen in Graves’ ophthalmopathy.  Clin Exp Immunol. 2005;  140 166-172
  • 9 Bednarczuk T, Stolarski C, Pawlik E, Slon M, Rowinski M, Kubota S, Hiromatsu Y, Bartoszewicz Z, Wall JR, Nauman J. Autoantibodies reactive with extracellular matrix proteins in patients with thyroid-associated ophthalmopathy.  Thyroid. 1999;  9 289-295
  • 10 De Bellis A, Sansone D, Coronella C, Conte M, Iorio S, Perrino S, Battaglia M, Bellastella G, Wall JR, Bellastella A, Bizzarro A. Serum antibodies to collagen XIII: a further good marker of active Graves' ophthalmopathy.  Clin Endocrinol (Oxf). 2005;  62 24-29
  • 11 Chang TC, Huang KM, Chang TJ, Lin SL. Correlation of orbital computed tomography and antibodies in patients with hyperthyroid Graves’ disease.  Clin Endocrinol (Oxf). 1990;  32 551-558
  • 12 Moncayo R, Baldissera I, Decristoforo C, Kendler D, Donnemiller E. Evaluation of immunological mechanisms mediating thyroid-associated ophthalmopathy by radionuclide imaging using the somatostatin analog 111In-octreotide.  Thyroid. 1997;  7 21-29
  • 13 Khalil HA, de Keizer RJ, Kijlstra A. Analysis of tear proteins in Graves’ ophthalmopathy by high performance liquid chromatography.  Am J Ophthalmol. 1988;  106 186-190
  • 14 Baker GR, Morton M, Rajapaska RS, Bullock M, Gullu S, Mazzi B, Ludgate M. Altered tear composition in smokers and patients with graves ophthalmopathy.  Arch Ophthalmol. 2006;  124 1451-1456
  • 15 Khalil HA, De Keizer RJ, Bodelier VM, Kijlstra A. Secretory IgA and lysozyme in tears of patients with Graves’ ophthalmopathy.  Doc Ophthalmol. 1989;  72 329-334
  • 16 Koo BS, Lee DY, Ha HS, Kim JC, Kim CW. Comparative analysis of the tear protein expression in blepharitis patients using two-dimensional electrophoresis.  J Proteome Res. 2005;  4 719-724
  • 17 Grus FH, Augustin AJ. Analysis of tear protein patterns by a neural network as a diagnostical tool for the detection of dry eyes.  Electrophoresis. 1999;  20 875-880
  • 18 Grus FH, Sabuncuo P, Herber S, Augustin AJ. Analysis of tear protein patterns for the diagnosis of dry eye.  Adv Exp Med Biol. 2002;  506 1213-1216
  • 19 Grus FH, Sabuncuo P, Augustin AJ. Analysis of tear protein patterns of dry-eye patients using fluorescent staining dyes and two-dimensional quantification algorithms.  Electrophoresis. 2001;  22 1845-1850
  • 20 Grus FH, Sabuncuo P, Augustin AJ. Quantitative analysis of tear protein profile for soft contact lenses--a clinical study].  Klin Monatsbl Augenheilkd. 2001;  218 239-242
  • 21 Grus FH, Sabuncuo P, Augustin A, Pfeiffer N. Effect of smoking on tear proteins.  Graefes Arch Clin Exp Ophthalmol. 2002;  240 889-892
  • 22 Stolwijk TR, Kuizenga A, van Haeringen NJ, Kijlstra A, Oosterhuis JA, van Best JA. Analysis of tear fluid proteins in insulin-dependent diabetes mellitus.  Acta Ophthalmol (Copenh). 1994;  72 357-362
  • 23 Herber S, Grus FH, Sabuncuo P, Augustin AJ. Changes in the tear protein patterns of diabetic patients using two-dimensional electrophoresis.  Adv Exp Med Biol. 2002;  506 623-626
  • 24 Dogru M, Katakami C, Inoue M. Tear function and ocular surface changes in noninsulin-dependent diabetes mellitus.  Ophthalmology. 2001;  108 586-592
  • 25 Herber S, Grus FH, Sabuncuo P, Augustin AJ. Two-dimensional analysis of tear protein patterns of diabetic patients.  Electrophoresis. 2001;  22 1838-1844
  • 26 Evans V, Vockler C, Friedlander M, Walsh B, Willcox MD. Lacryglobin in human tears, a potential marker for cancer.  Clin Experiment Ophthalmol. 2001;  29 161-163
  • 27 Grus FH, Augustin AJ. High performance liquid chromatography analysis of tear protein patterns in diabetic and non-diabetic dry-eye patients.  Eur J Ophthalmol. 2001;  11 19-24
  • 28 Qu Y, Adam BL, Yasui Y, Ward MD, Cazares LH, Schellhammer PF, Feng Z, Semmes OJ, Wright Jr GL, . Boosted decision tree analysis of surface-enhanced laser desorption/ionization mass spectral serum profiles discriminates prostate cancer from noncancer patients.  Clin Chem. 2002;  48 1835-1843
  • 29 Li J, Zhang Z, Rosenzweig J, Wang YY, Chan DW. Proteomics and bioinformatics approaches for identification of serum biomarkers to detect breast cancer.  Clin Chem. 2002;  48 1296-1304
  • 30 Petricoin EF, Ardekani AM, Hitt BA, Levine PJ, Fusaro VA, Steinberg SM, Mills GB, Simone C, Fishman DA, Kohn EC, Liotta LA. Use of proteomic patterns in serum to identify ovarian cancer.  Lancet. 2002;  359 572-577
  • 31 Rai AJ, Stemmer PM, Zhang Z, Adam BL, Morgan WT, Caffrey RE, Podust VN, Patel M, Lim LY, Shipulina NV, Chan DW, Semmes OJ, Leung HC. Analysis of Human Proteome Organization Plasma Proteome Project (HUPO PPP) reference specimens using surface enhanced laser desorption/ionization-time of flight (SELDI-TOF) mass spectrometry: multi-institution correlation of spectra and identification of biomarkers.  Proteomics. 2005;  5 3467-3474
  • 32 Roelofsen H, Alvarez-Llamas G, Schepers M, Landman K, Vonk RJ. Proteomics profiling of urine with surface enhanced laser desorption/ionization time of flight mass spectrometry.  Proteome Sci. 2007;  5 2
  • 33 Schipper R, Loof A, de Groot J, Harthoorn L, van Heerde W, Dransfield E. Salivary protein/peptide profiling with SELDI-TOF-MS.  Ann N Y Acad Sci. 2007;  1098 498-503
  • 34 Tomosugi N, Kitagawa K, Takahashi N, Sugai S, Ishikawa I. Diagnostic potential of tear proteomic patterns in Sjogren's syndrome.  J Proteome Res. 2005;  4 820-825
  • 35 Hida RY, Ohashi Y, Takano Y, Dogru M, Goto E, Fujishima H, Saito I, Saito K, Fukase Y, Tsubota K. Elevated levels of human alpha -defensin in tears of patients with allergic conjunctival disease complicated by corneal lesions: detection by SELDI ProteinChip system and quantification.  Curr Eye Res. 2005;  30 723-730
  • 36 Zhou L, Huang LQ, Beuerman RW, Grigg ME, Li SF, Chew FT, Ang L, Stern ME, Tan D. Proteomic analysis of human tears: defensin expression after ocular surface surgery.  J Proteome Res. 2004;  3 410-416
  • 37 Grus FH, Podust VN, Bruns K, Lackner K, Fu S, Dalmasso EA, Wirthlin A, Pfeiffer N. SELDI-TOF-MS ProteinChip array profiling of tears from patients with dry eye.  Invest Ophthalmol Vis Sci. 2005;  46 863-876
  • 38 Domberg J, Liu C, Papewalis C, Pfleger C, Xu K, Willenberg HS, Hermsen D, Scherbaum WA, Schloot NC, Schott M. Circulating chemokines in patients with autoimmune thyroid diseases.  Hormone and metabolic research Hormon- und Stoffwechselforschung. 2008;  40 416-421
  • 39 Liu C, Papewalis C, Domberg J, Scherbaum WA, Schott M. Chemokines and autoimmune thyroid diseases.  Horm Metab Res. 2008;  40 361-368
  • 40 Poon TC, Hui AY, Chan HL, Ang IL, Chow SM, Wong N, Sung JJ. Prediction of liver fibrosis and cirrhosis in chronic hepatitis B infection by serum proteomic fingerprinting: a pilot study.  Clin Chem. 2005;  51 328-335
  • 41 Dayal B, Ertel NH. ProteinChip technology: a new and facile method for the identification and measurement of high-density lipoproteins apoA-I and apoA-II and their glycosylated products in patients with diabetes and cardiovascular disease.  J Proteome Res. 2002;  1 375-380
  • 42 Grus FH, Sabuncuo P, Dick HB, Augustin AJ, Pfeiffer N. Changes in the tear proteins of diabetic patients.  BMC Ophthalmol. 2002;  2 4
  • 43 Mourits MP, Prummel MF, Wiersinga WM, Koornneef L. Clinical activity score as a guide in the management of patients with Graves’ ophthalmopathy.  Clin Endocrinol (Oxf). 1997;  47 9-14
  • 44 MacDermott AM. Defensins and other antimicrobial peptides at the ocular surface.  Ocul Surf. 2004;  2 229-247

Correspondence

Prof. Dr. G. J. Kahaly

Thyroid Research Laboratory

Department of Medicine I

Gutenberg University Hospital

Langenbeckstr. 1

55131 Mainz

Germany

Phone: +49/6131/17 37 68

Fax: +49/6131/17 34 60

Email: kahaly@ukmainz.de