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Longitudinal Multi-Omics Profiling of Aqueous Humor Implicates GALNS Depletion as a Pro-Fibrotic Mediator of

Yujuan Huang1,2, Mi Gui1, Xiangbin Kong2

  • 1Guangdong Eye Institute, Department of Ophthalmology, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, People's Republic of China.

Investigative Ophthalmology & Visual Science
|May 18, 2026
PubMed
Abstract

Insights

Anti-vascular endothelial growth factor (VEGF) therapy for proliferative diabetic retinopathy (PDR) may worsen fibrosis. This study identified GALNS depletion as a potential pro-fibrotic marker after anti-VEGF treatment in PDR patients.

Area of Science:

  • Ophthalmology
  • Molecular Biology
  • Biochemistry

Background:

  • Preoperative anti-vascular endothelial growth factor (VEGF) therapy in proliferative diabetic retinopathy (PDR) can lead to fibrovascular contraction, complicating treatment.
  • Understanding the molecular changes induced by anti-VEGF therapy is crucial for managing PDR complications.

Purpose of the Study:

  • To characterize proteo-metabolomic alterations following anti-VEGF therapy in PDR.
  • To validate potential molecular candidates associated with PDR progression or improvement.
  • To investigate the functional role of key molecules in fibrosis.

Main Methods:

  • Aqueous humor samples from PDR patients were analyzed using data-independent acquisition (DIA) proteomics and widely targeted metabolomics before and after anti-VEGF (aflibercept) treatment.
  • Molecules showing significant changes were classified as 'aggravation-type' or 'improvement-type'.
  • Validation was performed using parallel reaction monitoring (PRM) and selected reaction monitoring (SRM) in an independent cohort, with functional assessment of GALNS via siRNA knockdown.

Main Results:

  • Thirty-eight molecules, including 5 proteins and 33 metabolites, were identified. Three molecules (CAST, GALNS, 3-hydroxypropanoic acid [3-HPA]) were classified as 'aggravation-type'.
  • Only GALNS was robustly validated across cross-sectional and longitudinal analyses, showing depletion post-treatment in PDR patients with tractional retinal detachment.
  • GALNS knockdown in fibroblasts increased pro-fibrotic markers (α-SMA, collagen I), enhanced cell migration, and promoted contractility.

Conclusions:

  • GALNS depletion is identified as a potential pro-fibrotic molecule associated with anti-VEGF therapy in PDR.
  • GALNS may serve as a candidate biomarker for perioperative risk stratification in PDR patients undergoing anti-VEGF treatment.
  • CAST and 3-HPA require further investigation in larger cohorts.

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