PXD075862-1

PXD075862 is an original dataset announced via ProteomeXchange.

Title	Association of IL-8 and IL-18 Expression with Histone-Associated NETosis in Aqueous-Deficient and Evaporative Dry Eye Disease
Description	Purpose: Dry eye disease (DED) is a chronic inflammatory disorder of the ocular surface; however, the upstream immune mechanisms contributing to DED are complex and unclear. This study aimed to identify differentially expressed tear fluid proteins in patients with DED and elucidate the underlying disease mechanism. Methods: Tear samples from healthy controls (n=26), patients with aqueous-deficient DED (ADDE; n=29), and evaporative DED (EDE; n=26) were collected using Schirmer strips. Proteomic profiling was performed using nano-ESI LC-MS/MS (Orbitrap Eclipse) and analyzed with Proteome Discoverer against the UniProt human database (PSM and FDR ≤0.01). Differential proteins were identified using LIMMA and MBQN in R, followed by Gene Ontology-based pathway enrichment and network visualization. Selected candidates were validated by ELISA (IL-8, IL-18, histones), western blotting for citrullinated H3, and PicoGreen quantification of extracellular DNA. In vitro studies were performed using human corneal epithelial cells to further assess the expression of inflammatory and histone-associated markers. Results: A total of 2,596 tear proteins were identified across the groups, of which 101 were significantly altered (P ≤ 0.001). PCA and heatmaps revealed differential expression in healthy and dry eye tears, demonstrating elevated levels of inflammatory and oxidative stress proteins (MMP9, MAPK3, S100A8/A9, and SOD1) and a reduced abundance of lactoferrin, lysozyme, lipocalin-1, and mucins. Pathway enrichment analyses revealed the activation of neutrophil-mediated inflammation and extracellular chromatin release. Validation studies confirmed significant upregulation of neutrophil degranulation and NETosis-associated markers, including IL-8 (P≤ 0.01), IL-18 (P≤ 0.001), and histone proteins (H1, H2A/B, H3, and H4) (P≤ 0.001), confirming active NETosis in both dry eye subtypes compared with healthy controls. In vitro gene expression analysis confirmed a significant increase in histone and inflammatory markers. Conclusions: This study elucidates that proteomic profiling of tears in patients with DED, followed by targeted validation, reveals the activation of neutrophil-mediated inflammation and histone-associated NETosis in both aqueous-deficient and evaporative forms of the disease. These findings underscore the potential of neutrophil-derived and histone-associated proteins as tear biomarkers and contributors to ocular surface inflammation in DED.
HostingRepository	PRIDE
AnnounceDate	2026-09-07
AnnouncementXML	Submission_2026-09-07_04:21:05.601.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Vivek Singh
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2026-03-19 08:47:11	ID requested
⏵ 1	2026-09-07 04:21:06	announced

Prasad D, Siddiqui MG, Salman M, Sonar R, Maity M, Singh S, Donthineni PR, Basu S, Singh V, Association of IL-8 and IL-18 Expression With Histone-Associated NETosis in Aqueous-Deficient and Evaporative Dry Eye Disease. Invest Ophthalmol Vis Sci, 67(10):8(2026) [pubmed]

10.1167/iovs.67.10.8;

submitter keyword: tear proteomics, neutrophil degranulation, histones,ocular surface, dry eye, and inflammation

Dr. Vivek Singh
contact affiliation	Centre for Ocular Regeneration (CORE), Prof. Brien Holden Eye Research Centre (BHERC), L V Prasad Eye Institute, Hyderabad, Telangana, India
contact email	viveksingh@lvpei.org
lab head
Vivek Singh
contact affiliation	Prof. Brien Holden Eye Research Centre, L V Prasad Eye Institute
contact email	viveksingh@lvpei.org
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD075862
     1. Label: PRIDE project
     2. Name: Association of IL-8 and IL-18 Expression with Histone-Associated NETosis in Aqueous-Deficient and Evaporative Dry Eye Disease