Tear Proteome Analysis in Ocular Surface Diseases Using Label-free LC-MS/MS and Multiplexed-microarray Biomarker Validation

Overview

Journal Sci Rep

Specialty Science

Date 2017 Dec 14

PMID 29234088

Citations 51

Authors

Javier Soria

Arantxa Acera

Jesus Merayo-Lloves

Juan A Duran

Nerea Gonzalez

Sandra Rodriguez

Nikitas Bistolas

Soeren Schumacher

Frank F Bier

Harald Peter

Walter Stocklein

Tatiana Suarez

Affiliations

Soon will be listed here.

Abstract

We analyzed the tear film proteome of patients with dry eye (DE), meibomian gland dysfunction (MGD), and normal volunteers (CT). Tear samples were collected from 70 individuals. Of these, 37 samples were analyzed using spectral-counting-based LC-MS/MS label-free quantitation, and 33 samples were evaluated in the validation of candidate biomarkers employing customized antibody microarray assays. Comparative analysis of tear protein profiles revealed differences in the expression levels of 26 proteins, including protein S100A6, annexin A1, cystatin-S, thioredoxin, phospholipase A2, antileukoproteinase, and lactoperoxidase. Antibody microarray validation of CST4, S100A6, and MMP9 confirmed the accuracy of previously reported ELISA assays, with an area under ROC curve (AUC) of 87.5%. Clinical endpoint analysis showed a good correlation between biomarker concentrations and clinical parameters. In conclusion, different sets of proteins differentiate between the groups. Apolipoprotein D, S100A6, S100A8, and ceruloplasmin discriminate best between the DE and CT groups. The differences between antileukoproteinase, phospholipase A2, and lactoperoxidase levels allow the distinction between MGD and DE, and the changes in the levels of annexin A1, clusterin, and alpha-1-acid glycoprotein 1, between MGD and CT groups. The functional network analysis revealed the main biological processes that should be examined to identify new candidate biomarkers and therapeutic targets.

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References

de Paiva C, Villarreal A, Corrales R, Rahman H, Chang V, Farley W . Dry eye-induced conjunctival epithelial squamous metaplasia is modulated by interferon-gamma. Invest Ophthalmol Vis Sci. 2007; 48(6):2553-60. DOI: 10.1167/iovs.07-0069. View

Matheis N, Grus F, Breitenfeld M, Knych I, Funke S, Pitz S . Proteomics Differentiate Between Thyroid-Associated Orbitopathy and Dry Eye Syndrome. Invest Ophthalmol Vis Sci. 2015; 56(4):2649-56. DOI: 10.1167/iovs.15-16699. View

Li B, Sheng M, Li J, Yan G, Lin A, Li M . Tear proteomic analysis of Sjögren syndrome patients with dry eye syndrome by two-dimensional-nano-liquid chromatography coupled with tandem mass spectrometry. Sci Rep. 2014; 4:5772. PMC: 4145314. DOI: 10.1038/srep05772. View

Lin C, Chin C, Wu H, Chen S, Ho C, Ko M . Hubba: hub objects analyzer--a framework of interactome hubs identification for network biology. Nucleic Acids Res. 2008; 36(Web Server issue):W438-43. PMC: 2447731. DOI: 10.1093/nar/gkn257. View

Mackie I, Seal D . Diagnostic implications of tear protein profiles. Br J Ophthalmol. 1984; 68(5):321-4. PMC: 1040331. DOI: 10.1136/bjo.68.5.321. View

Zhu W, Smith J, Huang C . Mass spectrometry-based label-free quantitative proteomics. J Biomed Biotechnol. 2009; 2010:840518. PMC: 2775274. DOI: 10.1155/2010/840518. View

Braisted J, Kuntumalla S, Vogel C, Marcotte E, Rodrigues A, Wang R . The APEX Quantitative Proteomics Tool: generating protein quantitation estimates from LC-MS/MS proteomics results. BMC Bioinformatics. 2008; 9:529. PMC: 2639435. DOI: 10.1186/1471-2105-9-529. View

Keller A, Nesvizhskii A, Kolker E, Aebersold R . Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search. Anal Chem. 2002; 74(20):5383-92. DOI: 10.1021/ac025747h. View

Funke S, Azimi D, Wolters D, Grus F, Pfeiffer N . Longitudinal analysis of taurine induced effects on the tear proteome of contact lens wearers and dry eye patients using a RP-RP-Capillary-HPLC-MALDI TOF/TOF MS approach. J Proteomics. 2012; 75(11):3177-90. DOI: 10.1016/j.jprot.2012.03.018. View

10.

Uchino Y, Mauris J, Woodward A, Dieckow J, Amparo F, Dana R . Alteration of galectin-3 in tears of patients with dry eye disease. Am J Ophthalmol. 2015; 159(6):1027-1035.e3. PMC: 4426220. DOI: 10.1016/j.ajo.2015.02.008. View

11.

Csosz E, Boross P, Csutak A, Berta A, Toth F, Poliska S . Quantitative analysis of proteins in the tear fluid of patients with diabetic retinopathy. J Proteomics. 2012; 75(7):2196-204. DOI: 10.1016/j.jprot.2012.01.019. View

12.

Boehm N, Funke S, Wiegand M, Wehrwein N, Pfeiffer N, Grus F . Alterations in the tear proteome of dry eye patients--a matter of the clinical phenotype. Invest Ophthalmol Vis Sci. 2013; 54(3):2385-92. DOI: 10.1167/iovs.11-8751. View

13.

Lopez-Miguel A, Teson M, Martin-Montanez V, Enriquez-de-Salamanca A, Stern M, Gonzalez-Garcia M . Clinical and Molecular Inflammatory Response in Sjögren Syndrome-Associated Dry Eye Patients Under Desiccating Stress. Am J Ophthalmol. 2015; 161:133-41.e1-2. DOI: 10.1016/j.ajo.2015.09.039. View

14.

Gomes J, Azar D, Baudouin C, Efron N, Hirayama M, Horwath-Winter J . TFOS DEWS II iatrogenic report. Ocul Surf. 2017; 15(3):511-538. DOI: 10.1016/j.jtos.2017.05.004. View

15.

Tseng S . Staging of conjunctival squamous metaplasia by impression cytology. Ophthalmology. 1985; 92(6):728-33. DOI: 10.1016/s0161-6420(85)33967-2. View

16.

Dezso Z, Oltvai Z, Barabasi A . Bioinformatics analysis of experimentally determined protein complexes in the yeast Saccharomyces cerevisiae. Genome Res. 2003; 13(11):2450-4. PMC: 403764. DOI: 10.1101/gr.1073603. View

17.

Craig J, Tomlinson A . Importance of the lipid layer in human tear film stability and evaporation. Optom Vis Sci. 1997; 74(1):8-13. DOI: 10.1097/00006324-199701000-00014. View

18.

Mackie I, Seal D . Confirmatory tests for the dry eye of Sjögren's syndrome. Scand J Rheumatol Suppl. 1986; 61:220-3. View

19.

Wu G, Feng X, Stein L . A human functional protein interaction network and its application to cancer data analysis. Genome Biol. 2010; 11(5):R53. PMC: 2898064. DOI: 10.1186/gb-2010-11-5-r53. View

20.

Tong L, Zhou L, Beuerman R, Zhao S, Li X . Association of tear proteins with Meibomian gland disease and dry eye symptoms. Br J Ophthalmol. 2010; 95(6):848-52. DOI: 10.1136/bjo.2010.185256. View