Bottom–up protein identifications from microliter quantities of individual human tear samples. Important steps towards clinical relevance.

Raus, Peter; Raghuraman, Bharath Kumar; Pinkse, Martijn W. H.; Verhaert, Peter

doi:10.1016/j.euprot.2015.06.005

Cited by 3 publications

(4 citation statements)

References 12 publications

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“…Considering the overlap between untargeted and targeted analyses, a final list of 124 putative biomarkers were selected, based on criteria of having p -values <0.05 and fold change higher than |1.5|. Four additional proteins were considered of interest for further investigation based on their relevance to SS-DED in literature. − …”

Section: Resultsmentioning

confidence: 99%

Discovery and Verification of Sjögren’s Syndrome Protein Biomarkers in Tears by Targeted LC-MRM

Lépine,

Robert,

Sleno

2024

J. Proteome Res.

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Sjogren's syndrome (SS) is an autoimmune rheumatic disorder characterized by exocrine gland dysfunction, mainly from the lacrimal and salivary glands. The disease causes severe aqueous dry eye syndrome (DED) and is associated with high rates of complications, including corneal ulceration, scaring, and perforation. Systemic complications may occur as well as a higher risk of developing lymphoma. Diagnosis of SS-DED is often delayed and difficult to establish. With the aim of discovering biomarkers to help discriminate SS-DED patients, a combination of untargeted and targeted LC-MS/MS analyses were performed on tear samples collected on Schirmer strips and subjected to tryptic digestion. Following the analysis of three cohorts and the development of two targeted LC-sMRM methods for the verification of putative biomarkers found in the first cohort of samples, 64 proteins could be linked to Sjogren's syndrome, in the hopes of helping to confirm diagnoses as well as potentially stratifying the severity of disease in these patients. Proteins that were increased in SS-DED showed activation of the immune system and alterations in homeostasis. Several proteases and protease inhibitors were found to be significantly changing in SS-DED, as well as a consistent decrease in specific proteins known to be secreted by the lacrimal gland.

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Section: Resultsmentioning

confidence: 99%

Discovery and Verification of Sjögren’s Syndrome Protein Biomarkers in Tears by Targeted LC-MRM

Lépine,

Robert,

Sleno

2024

J. Proteome Res.

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“…We opted for Schirmer test strips instead of glass capillaries or conjunctival swabs as most convenient clinician and patient friendly tear sampling tool. 20 The advantage of glass capillary tubes is that it does not induce local irritation and minimises transudation; however, it can be used only by trained personnel in cooperative patients. 21 Schirmer strips are a non-invasive body fluid collection method commonly used in ophthalmological practice for testing the severity of dry eyes.…”

Section: Discussionmentioning

confidence: 99%

SARS-CoV-2 RNA and antibodies in tear fluid

et al. 2021

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Background/aimsSARS-CoV-2 is highly contagious. More evidence concerning extrapulmonary transmission routes such as the eyes is urgently needed. Although the humoral immune response is important in the viral containment, the local response in tears has not yet been studied. The aim of our study was twofold: to assess the prevalence of both SARS-CoV-2 RNA and antibodies in tear fluid.MethodsIn a first series, nasopharyngeal sampling and tear sampling by Schirmer test strips were performed in 26 acutely ill patients with COVID-19 to assess the presence of SARS-CoV-2 RNA by reverse transcription PCR. In a second series, IgG and IgA responses to SARS-CoV-2 spike protein in serum and tear fluid of convalescent individuals (n=22) were compared with control individuals (n=15) by ELISA.ResultsSARS-CoV-2 RNA was detected in tears of 7/26 (26.9%) patients with COVID-19. None of them had ocular symptoms. Convalescent individuals displayed a significant higher ratio of IgG (p<0.0001) and IgA (p=0.0068) in tears compared with control individuals. A sensitivity of 77.3% and specificity of 93.3% was observed for IgG, and 59.1% and 100% for IgA.ConclusionsOur results demonstrate the presence of SARS-CoV-2 RNA and a local IgG and IgA immune response in tear fluid. These data confirm the possibility of SARS-CoV-2 transmission through tear fluid and the importance of the eye as a first defence against SARS-CoV-2, indicating the potential of tears as a non-invasive surrogate for serum in monitoring the host immune response.

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“…The tear film protects the eye against infection and ensures adequate lubrication and nutrition of the ocular surface . Proteins in tears, mainly contained in the aqueous phase, play a central role where a change in the quantity and quality of proteins at the ocular surface has previously demonstrated correlation with ocular disease. ,− …”

Section: Introductionmentioning

confidence: 99%

“… 5 Proteins in tears, mainly contained in the aqueous phase, play a central role where a change in the quantity and quality of proteins at the ocular surface has previously demonstrated correlation with ocular disease. 1 , 6 − 10 …”

Section: Introductionmentioning

confidence: 99%

Targeted Workflow Investigating Variations in the Tear Proteome by Liquid Chromatography Tandem Mass Spectrometry

Lépine,

Zambito,

Sleno

2023

ACS Omega

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Proteins in tears have an important role in eye health and have been shown as a promising source of disease biomarkers. The goal of this study was to develop a robust, sensitive, and targeted method for profiling tear proteins to examine the variability within a group of healthy volunteers over three days. Inter-individual and inter-day variabilities were examined to contribute to understanding the normal variations in the tear proteome, as well as to establish which proteins may be better candidates as eventual biomarkers of specific diseases. Tear samples collected on Schirmer strips were subjected to bottom-up proteomics, and resulting peptides were analyzed using an optimized targeted method measuring 226 proteins by liquid chromatography-scheduled multiple reaction monitoring. This method was developed using an in-house database of identified proteins from tears compiled from high-resolution data-dependent liquid chromatography tandem mass spectrometry data. The measurement of unique peptide signals can help better understand the dynamics of each of these proteins in tears. Some interesting trends were seen in specific pathways or protein classes, including higher variabilities for those involved in glycolysis, glutathione metabolism, and cytoskeleton proteins and lower variation for those involving the degradation of the extracellular matrix. The overall aim of this study was to contribute to the field of tear proteomics with the development of a novel and targeted method that is highly amenable to the clinical laboratory using high flow LC and commonly used triple quadrupole mass spectrometry while ensuring that protein quantitation was reported based on unique peptides for each protein and robust peak areas with data normalization. These results report on variabilities on over 200 proteins that are robustly detected in tear samples from healthy volunteers with a simple sample preparation procedure.

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Bottom–up protein identifications from microliter quantities of individual human tear samples. Important steps towards clinical relevance.

Cited by 3 publications

References 12 publications

Discovery and Verification of Sjögren’s Syndrome Protein Biomarkers in Tears by Targeted LC-MRM

Discovery and Verification of Sjögren’s Syndrome Protein Biomarkers in Tears by Targeted LC-MRM

SARS-CoV-2 RNA and antibodies in tear fluid

Targeted Workflow Investigating Variations in the Tear Proteome by Liquid Chromatography Tandem Mass Spectrometry

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