Tear proteomics in evaporative dry eye disease

Versura, Piera; Nanni, Paolo; Bavelloni, Alberto; Blalock, William L.; Piazzi, Manuela; Roda, Aldo; Campos, Emilio C.

doi:10.1038/eye.2010.7

Cited by 138 publications

(123 citation statements)

References 26 publications

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“…20 A statistically significant decrease in lactoferrin, lipophilin, and lipocalin amount was found in patients versus controls; however, our study did not show a change in lysozyme and zinc-alpha-2-glycoprotein amounts. Tear proteomics on contact lens wearers, 42 Sjögren's syndrome patients, 13,[16][17][18] and keratoconus patients 43,44 have shown upregulation and down-regulation of unique proteins.…”

Section: Discussioncontrasting

confidence: 50%

“…6,19 Versura et al, in a recent study, showed that the tear protein changes anticipate the onset of more extensive clinical signs in early stage DE disease. 20 Changes of tear protein profile also have been shown to correlate with DE severity, 13 and the levels of certain proteins have been correlated with the severity of meibomian gland disease. 14 Various factors can influence tear proteomics, including the tear collection methods, 7,[21][22][23] storage, and analysis techniques.…”

Section: Resultsmentioning

confidence: 99%

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iTRAQ Quantitative Proteomics in the Analysis of Tears in Dry Eye Patients

Srinivasan

Thangavelu

Zhang³

et al. 2012

Invest. Ophthalmol. Vis. Sci.

108

116

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PURPOSE. We analyzed the change in protein expression of tear film proteins in dry eye (DE) and non-DE (NDE) patients using isobaric tag for relative and absolute quantitation (iTRAQ) technology.METHODS. We categorized 24 participants into NDE, and mild (MDE), moderate-to-severe (MSDE), and mixed (MXDE) DE on the basis of clinical DE tests. Tear samples (n ¼ 6 subjects/ group) were collected using Schirmer's strips. Proteins were extracted from strips and were quantified using the Bradford assay. Protein from each sample was pooled as internal standard (IS), and 20 lg protein from each sample and the IS were digested and labeled with different tandem mass tag (TMT) isobaric mass tag labeling reagent. The reaction was quenched and the labeled peptides were mixed. Samples were injected for liquid chromatography-mass spectrometry (LC/MS/ MS) analysis on the Orbitrap mass spectrometer. Bioinformatic analyses were performed using protein information resource (PIR). RESULTS.Combined results showed a total of 386 proteins in tears as determined by the iTRAQ experiments. An average of 163 proteins was detected in each of 6 biologic replicates. Of those, 55% were detected 6 times and 90% were detected multiple times (>2). In addition to the down-regulation of commonly reported proteins, such as lipocalin-1, lysozyme, and prolactin-inducible protein across all sub groups of DE, a number of proteins were significantly differentially regulated in MSDE and other subgroups of DE. A greater number of proteins were down-regulated in MSDE versus MDE, and the specific functions involved include response to stimulus (8 vs. 6 proteins), immune system process (6 vs. 4), regulation of biologic processes (3 vs. 3), and ion transport (2 vs. 2).CONCLUSIONS. iTRAQ is one of the newest tools for quantitative mass spectrometry in tear proteome research. Differences in the protein ratios can be detected between normal and DE patients. PIR is a useful resource to interpret pathways and functions of proteins. (Invest Ophthalmol Vis Sci. 2012; 53:5052-5059) DOI:10.1167/iovs.11-9022 T he Dry Eye Workshop in 2007 defined dry eye (DE) as a multifactorial ocular surface disease diagnosed by symptoms of discomfort, and signs of visual disturbance, tear film instability, and ocular surface damage, accompanied by increased osmolarity of the tear film and ocular surface inflammation.1 It is evident from the definition that the tear film and ocular surface are altered in DE disease. The tear film serves/performs a variety of functions and is composed of various substances, including proteins, lipids, mucins, salts, and other organic molecules.2 The aqueous component constitutes the majority of the tear layer, and the proteins in the tear film are believed to have a key role in the protection of the external surface from potential pathogens, and also are involved in modulation of wound healing process. [3][4][5][6] The major source of tear proteins is the secretory acinar cells of the lacrimal gland, including the primary proteins of the tear film: lyso...

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Section: Discussioncontrasting

confidence: 50%

Section: Resultsmentioning

confidence: 99%

iTRAQ Quantitative Proteomics in the Analysis of Tears in Dry Eye Patients

Srinivasan

Thangavelu

Zhang³

et al. 2012

Invest. Ophthalmol. Vis. Sci.

108

116

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“…For example, increased abundance of ZAG was found in the tears of keratoconus patients [15], in patients with Grave's ophthalmopathy [41] or keratitis [42]. On the other hand, no significant changes of this protein were noticed in tears from patients with dry eye syndrome [43]. In the present study we have found elevated levels of ZAG in aniridia tears (1.4-fold).…”

Section: Zinc-α2-glycoproteinsupporting

confidence: 52%

Analysis of protein composition and protein expression in the tear fluid of patients with congenital aniridia

Ihnatko

Edén

Lagali

et al. 2013

Journal of Proteomics

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Aniridia is a rare congenital genetic disorder caused by haploinsuffiency of the PAX6 gene, the master gene for development of the eye. The expression of tear proteins in aniridia is unknown. To screen for proteins involved in the aniridia pathophysiology, the tear fluid of patients with diagnosed congenital aniridia was examined using two-dimensional electrophoresis (2-DE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Two-dimensional map of tear proteins in aniridia has been established and 7 proteins were differentially expressed with P less than 0.01 between aniridia patients and control subjects. Five of them were more abundant in healthy subjects, particularly alpha-enolase, peroxiredoxin 6, cystatin S, gelsolin, apolipoprotein A-1 and two other proteins, zinc-alpha 2-glycoprotein and lactoferrin were more expressed in the tears of aniridia patients. Moreover, immunoblot analysis revealed elevated levels of vascular endothelial growth factor (VEGF) in aniridia tears which is in concordance with clinical finding of pathological blood and lymph vessels in the central and peripheral cornea of aniridia patients. The proteins with different expression in patients tears may be new candidate molecules involved in the pathophysiology of aniridia and thus may be helpful for development of novel treatment strategies for the symptomatic therapy of this vision threatening condition. Biological significance This study is first to demonstrate protein composition and protein expression in aniridic tears and identifies proteins with different abundance in tear fluid from patients with congenital aniridia vs. healthy tears

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“…30 Our results showed a decrease in the levels of lipophilin-A and -C in tears from KC patients vs controls as reported by Versura et al in dry eye patients. 31 Thus, decreased levels of lipophilin-A and -C in tears from KC patients may lead to altered tear stability due to higher levels of free lipid in tears. When the lipid layer is abnormal, tear film evaporates faster, which in turn leads to rapid tear dissociation times and finally to epithelial cell damage.…”

Section: Discussionmentioning

confidence: 99%

Changes in tear protein profile in keratoconus disease

Acera

Vecino

Rodríguez-Agirretxe³

et al. 2011

Eye

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Purpose To analyze tear protein profile variations in patients with keratoconus (KC) and to compare them with those of control subjects. Subjects and methods Tears from 12 normal subjects and 12 patients with KC were analyzed by two-dimensional gel electrophoresis (2-DE) and liquid chromatography-mass spectrometry (LC-MS). Analysis of the 2-DE gels was performed using Progenesis SameSpots software (Nonlinear Dynamics). Proteins exhibiting high variation in expression levels (P-value o0.05) were identified using matrix-assisted laser desorption/ionization-TOF spectrometry. For LC-MS analysis, a label-free quantification approach was used. Tears were digested with trypsin, subjected to data-independent acquisition (MS E ) analysis, and identified proteins were relatively quantified using ProteinLynx Global Server software (Waters). Results The 2-DE and LC-MS analyses revealed a significant decrease in the levels of members of the cystatin family and an increase in lipocalin-1 in KC patients. A 1.43-fold decrease was observed for cystatin-S by 2-DE, and 1.69-and 1.56-fold for cystatin-SN and cystatin-SA by LC-MS, respectively. The increase in lipocalin-1 was observed by both methods with fold changes of 1.26 in the 2-DE approach and 1.31 according to LC-MS. Significant protein upregulation was also observed for Ig-j chain C and Ig J chain proteins by 2-DE. Levels of lipophilin-C, lipophilin-A, and phospholipase A2 were decreased in tears from KC patients according to LC-MS. Serum albumin was found to be increased in KC patients according to LC-MS. Conclusion The results show differences in the tear protein profile of KC and control subjects. These changes are indicative of alterations in tear film stability and in interactions with the corneal surface in KC patients.

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Tear proteomics in evaporative dry eye disease

Cited by 138 publications

References 26 publications

iTRAQ Quantitative Proteomics in the Analysis of Tears in Dry Eye Patients

iTRAQ Quantitative Proteomics in the Analysis of Tears in Dry Eye Patients

Analysis of protein composition and protein expression in the tear fluid of patients with congenital aniridia

Changes in tear protein profile in keratoconus disease

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