Label-free quantitative proteomic analysis of aqueous humor in patients with pathologic myopia_Chinese Journal of Ocular Fundus Diseases

Authors：

Xue Min , Ke Yifeng , Ren Xinjun , Liu Juping , Fan Xiao’e ,  Li Xiaorong

Tianjin Key Laboratory of Retinal Functions and Diseases, Tianjin International Joint Research and Development Centre of Ophthalmology and Vision Science, Eye Institute and School of Optometry, Tianjin Medical University Eye Hospital, Tianjin 300384, China;
Xue Min is working at Department of Ophthalmology of Anhui NO.2 Provincial People’s Hospital, Hefei 230041, China;

Corresponding author：

Li Xiaorong, Email: xiaorli@163.com

Keywords：

Myopia, degenerative; Aqueous humor; Proteomics

DOI：

10.3760/cma.j.cn511434-20200907-00436

Video：

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Objective To characterize proteomic profile in aqueous humor of patients with pathologic myopia (PM) using quantitative proteomic analysis, which may provide new clues to understand the mechanisms and possible treatments of PM.Methods A cross-sectional study. From January 2019 to August 2019, aqueous humor samples (32 cataract patients) were collected for quantitative proteomic analysis using liquid chromatography tandem mass spectrometry at Tianjin Medical University Eye Hospital. There were 11 males and 21 females. They were 58-76 years old with an average age of 68.41±6.09 years old. Sixteen patients with PM were regarded as PM group, 16 patients without myopia were regarded as the control group. The aqueous humor samples (100-150 μl ) were collected from all patients before cataract surgery. Using protein quantification and non-labeled liquid chromatography tandem mass spectrometry analysis, differentially expressed proteins were obtained. Five different proteins were randomly selected for ELISA verification. The differentially expressed proteins were further analyzed by gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes, which were validated using ELISA in the other twenty samples of each group.Results A total of 583 proteins were identified and 101 proteins were found to be differentially expressed, including 63 up-regulated proteins and 38 down-regulated proteins. ELISA verification results showed that the expression trend of the 5 differentially expressed proteins between the PM group and the control group was consistent with the results of Label-free quantitative proteomics analysis. The main classifications of these differentially expressed proteins were protein-binding activity modulator, defense/immunity protein, protein modifying enzyme, metabolite interconversion enzyme, extracellular matrix protein, transfer/carrier protein and so on. The bioinformatics analysis suggested that PM was closely associated with inflammation and immune interactions, and remodeling of extracellular matrix.Conclusions Compared with the control group, the protein expression profile of PM patients' aqueous humor specimens has obvious changes. These differences indicate that PM is closely related to inflammation and immune interaction and extracellular matrix remodeling.

Citation： Xue Min, Ke Yifeng, Ren Xinjun, Liu Juping, Fan Xiao’e, Li Xiaorong. Label-free quantitative proteomic analysis of aqueous humor in patients with pathologic myopia. Chinese Journal of Ocular Fundus Diseases, 2020, 36(12): 936-942. doi: 10.3760/cma.j.cn511434-20200907-00436 Copy

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1. Neelam K, Cheung CM, Ohno-Matsui K, et al. Choroidal neovascularization in pathological myopia[J]. Prog Retin Eye Res, 2012, 31(5): 495-525. DOI: 10.1016/j.preteyeres.2012.04.001.
2. Curtin BJ. Physiologic vs pathologic myopia: genetics vs environment[J]. Ophthalmology, 1979, 86(5): 681-691. DOI: 10.1016/s0161-6420(79)35466-5.
3. Huang Y, Kee CS, Hocking PM, et al. A genome-wide association study for susceptibility to visual experience-induced myopia[J]. Invest Ophthalmol Vis Sci, 2019, 60(2): 559-569. DOI: 10.1167/iovs.18-25597.
4. Wei Q, Jiang C, Ye X, et al. Vitreous proteomics provides new insights into antivascular endothelial growth factor therapy for pathologic myopia choroid neovascularization[J]. J Interferon Cytokine Res, 2019, 39(12): 786-796. DOI: 10.1089/jir.2019.0030.
5. Tse JS, Lam TC, Cheung JK, et al. Data on assessment of safety and tear proteome change in response to orthokeratology lens-insight from integrating clinical data and next generation proteomics[J/OL]. Data Brief, 2020, 29: 105186[2020-01-28]. https://pubmed.ncbi.nlm.nih.gov/32071970/. DOI: 10.1016/j.dib.2020.105186.
6. Latosinska A, Vougas K, Makridakis M, et al. Comparative analysis of label-free and 8-plex iTRAQ approach for quantitative tissue proteomic analysis[J/OL]. PLoS One, 2015, 10(9): e0137048[2015-09-02]. https://pubmed.ncbi.nlm.nih.gov/26331617/. DOI: 10.1371/journal.pone.0137048.
7. Huang Q, Yang L, Luo J, et al. SWATH enables precise label-free quantification on proteome scale[J]. Proteomics, 2015, 15(7): 1215-1223. DOI: 10.1002/pmic.201400270.
8. Barbas-Bernardos C, Armitage EG, Garcia A, et al. Looking into aqueous humor through metabolomics spectacles-exploring its metabolic characteristics in relation to myopia[J]. J Pharm Biomed Anal, 2016, 127: 18-25. DOI: 10.1016/j.jpba.2016.03.032.
9. Ji Y, Rao J, Rong X, et al. Metabolic characterization of human aqueous humor in relation to high myopia[J]. Exp Eye Res, 2017, 159: 147-155. DOI: 10.1016/j.exer.2017.03.004.
10. Duan X, Lu Q, Xue P, et al. Proteomic analysis of aqueous humor from patients with myopia[J]. Mol Vis, 2008, 14: 370-377. DOI: 10.1016/j.moicatb.2007.10.003.
11. Wildschutz L, Ackermann D, Witten A, et al. Transcriptomic and proteomic analysis of iris tissue and aqueous humor in juvenile idiopathic arthritis-associated uveitis[J]. J Autoimmun, 2019, 100: 75-83. DOI: 10.1016/j.jaut.2019.03.004.
12. Yang Q, Lu H, Song X, et al. iTRAQ-based proteomics investigation of aqueous humor from patients with Coats' disease[J/OL]. PLoS One, 2016, 11(7): e0158611[2016-07-14]. https://pubmed.ncbi.nlm.nih.gov/27416065/. DOI: 10.1371/journal.pone.0158611.
13. Kang GY, Bang JY, Choi AJ, et al. Exosomal proteins in the aqueous humor as novel biomarkers in patients with neovascular age-related macular degeneration[J]. J Proteome Res, 2014, 13(2): 581-595. DOI: 10.1021/pr400751k.
14. Ohno-Matsui K, Kawasaki R, Jonas JB, et al. International photographic classification and grading system for myopic maculopathy[J]. Am J Ophthalmol, 2015, 159(5): 877-883. DOI: 10.1016/j.ajo.2015.01.022.
15. Qu SC, Xu D, Li TT, et al. iTRAQ-based proteomics analysis of aqueous humor in patients with dry age-related macular degeneration[J]. Int J Ophthalmol, 2019, 12(11): 1758-1766. DOI: 10.18240/ijo.2019.11.15.
16. Pollreisz A, Funk M, Breitwieser FP, et al. Quantitative proteomics of aqueous and vitreous fluid from patients with idiopathic epiretinal membranes[J]. Exp Eye Res, 2013, 108: 48-58. DOI: 10.1016/j.exer.2012.11.010.
17. Kim MS, Gu BH, Song S, et al. ITI-H4, as a biomarker in the serum of recurrent pregnancy loss (RPL) patients[J]. Mol Biosyst, 2011, 7(5): 1430-1440. DOI: 10.1039/c0mb00219d.
18. Ji Y, Rong X, Ye H, et al. Proteomic analysis of aqueous humor proteins associated with cataract development[J]. Clin Biochem, 2015, 48(18): 1304-1309. DOI: 10.1016/j.clinbiochem.2015.08.006.
19. Clark SJ, Bishop PN. The eye as a complement dysregulation hotspot[J]. Semin Immunopathol, 2018, 40(1): 65-74. DOI: 10.1007/s00281-017-0649-6.
20. Skeie JM, Mahajan VB. Proteomic landscape of the human choroid-retinal pigment epithelial complex[J]. JAMA Ophthalmol, 2014, 132(11): 1271-1281. DOI: 10.1001/jamaophthalmol.2014.2065.
21. Rada JA, Shelton S, Norton TT. The sclera and myopia[J]. Exp Eye Res, 2006, 82(2): 185-200. DOI: 10.1016/j.exer.2005.08.009.
22. Kano H, Kobayashi K, Herrmann R, et al. Deficiency of alpha-dystroglycan in muscle-eye-brain disease[J]. Biochem Biophys Res Commun, 2002, 291(5): 1283-1286. DOI: 10.1006/bbrc.2002.6608.
23. Jiao H, Manya H, Wang S, et al. Novel POMGnT1 mutations cause muscle-eye-brain disease in Chinese patients[J]. Mol Genet Genomics, 2013, 288(7-8): 297-308. DOI: 10.1007/s00438-013-0749-5.
24. Thapa N, Lee BH, Kim IS. TGFBIp/betaig-h3 protein: a versatile matrix molecule induced by TGF-beta[J]. Int J Biochem Cell Biol, 2007, 39(12): 2183-2194. DOI: 10.1016/j.biocel.2007.06.004.
25. Flitcroft DI, Loughman J, Wildsoet CF, et al. Novel myopia genes and pathways identified from syndromic forms of myopia[J]. Invest Ophthalmol Vis Sci, 2018, 59(1): 338-348. DOI: 10.1167/iovs.17-22173.
26. Wu WW, Molday RS. Defective discoidin domain structure, subunit assembly, and endoplasmic reticulum processing of retinoschisin are primary mechanisms responsible for X-linked retinoschisis[J]. J Biol Chem, 2003, 278(30): 28139-28146. DOI: 10.1074/jbc.M302464200.
27. Sauer CG, Gehrig A, Warneke-Wittstock R, et al. Positional cloning of the gene associated with X-linked juvenile retinoschisis[J]. Nat Genet, 1997, 17(2): 164-170. DOI: 10.1038/ng1097-164.
28. Weber BH, Schrewe H, Molday LL, et al. Inactivation of the murine X-linked juvenile retinoschisis gene, Rs1h, suggests a role of retinoschisin in retinal cell layer organization and synaptic structure[J]. Proc Natl Acad Sci USA, 2002, 99(9): 6222-6227. DOI: 10.1073/pnas.092528599.

Chinese Journal of Ocular Fundus Diseases

Label-free quantitative proteomic analysis of aqueous humor in patients with pathologic myopia

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