April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
The Potential Role of O-GlcNAcylation as Biomarker in Diabetic Retinopathy and Diabetic Macular Edema
Yasir J. Sepah; Ronald Copeland; Roomasa Channa; Junfeng Ma; Afsheen Khwaja; Elham Hatef; Mohamed Ibrahim; Diana V. Do; Gerald W. Hart; Quan D. Nguyen
Author Affiliations & Notes
  • Yasir J. Sepah
    The Wilmer Eye Institute, Baltimore, Maryland
  • Ronald Copeland
    Biological Chemistry, Johns Hopkins University, Baltimore, Maryland
  • Roomasa Channa
    The Wilmer Eye Institute, Baltimore, Maryland
  • Junfeng Ma
    Biological Chemistry, Johns Hopkins University, Baltimore, Maryland
  • Afsheen Khwaja
    The Wilmer Eye Institute, Baltimore, Maryland
  • Elham Hatef
    The Wilmer Eye Institute, Baltimore, Maryland
  • Mohamed Ibrahim
    The Wilmer Eye Institute, Baltimore, Maryland
  • Diana V. Do
    The Wilmer Eye Institute, Baltimore, Maryland
  • Gerald W. Hart
    Biological Chemistry, Johns Hopkins University, Baltimore, Maryland
  • Quan D. Nguyen
    The Wilmer Eye Institute, Baltimore, Maryland
  • Footnotes
    Commercial Relationships  Yasir J. Sepah, None; Ronald Copeland, None; Roomasa Channa, None; Junfeng Ma, None; Afsheen Khwaja, None; Elham Hatef, None; Mohamed Ibrahim, None; Diana V. Do, Genentech, Regeneron (F); Gerald W. Hart, None; Quan D. Nguyen, Genentech, Regeneron (F)
  • Footnotes
    Support  NIH-1R24DK084949-01
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 4457. doi:
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      Yasir J. Sepah, Ronald Copeland, Roomasa Channa, Junfeng Ma, Afsheen Khwaja, Elham Hatef, Mohamed Ibrahim, Diana V. Do, Gerald W. Hart, Quan D. Nguyen; The Potential Role of O-GlcNAcylation as Biomarker in Diabetic Retinopathy and Diabetic Macular Edema. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4457.

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Abstract

Purpose: : O-linked b-N-acetylglucosamine (O-GlcNAc) is a post-translational modification consisting of addition of a single GlcNAc moiety to serine and/or threonine residues of numerous nuclear and cytoplasmic proteins. Recent data supports the hypothesis that a hyperglycemia-induced increase in O-GlcNAcylation underlies insulin resistance and glucose toxicity. O-GlcNAc has not been established on secreted proteins. Presence of such proteins in the ocular fluid (OF) of eyes with DR/DME has also not been investigated. We aim to establish the presence of protein O-GlcNAcylation in OF of patients with diabetic retinopathy (DR) and diabetic macular edema (DME).

Methods: : Anterior chamber (AC) fluid was collected from one eye of each of 25 subjects with DME and non-proliferative or inactive proliferative DR who are participating in a clinical trial for DME. The eyes have not been treated with anti-VEGF/steroid therapies for at least 3 months prior to sample collection. AC fluids samples were stored at -80C°. Proteins contained in AC fluid (approx. 10 µl) was subjected to SDS-PAGE analysis and transferred onto polyvinylidenefluroride (PVDF) membranes. Membranes were blocked in Tris-buffer saline with Tween (0.1%, [v/v] Tween 20) containing 3% BSA followed by overnight blotting with O-GlcNAc specific antibody. To test for specificity, the O-GlcNAc antibody was preincubated with free GlcNAc for 1h prior incubation with PVDF membrane. Detection was performed by enhanced chemiluminescence.

Results: : Immunoblotting demonstrated that at least 4 protein bands were reactive to an O-GlcNAc specific antibody in all 25 samples. Immunoreactivity of the antibody could be competed away with GlcNAc, suggesting that proteins contained within the AC of eyes with DR and DME are O-GlcNAcylated. The identity of O-GlcNAcylated proteins is being confirmed using mass spectrometry.

Conclusions: : The index study has shown the presence of O-GlcNAcylated proteins in AC fluid of eyes with DR and DME, which has not been illustrated previously. Results from our novel study suggest that O-GlcNAcylation may be a "sensor" that could be employed to predict the various clinical stages and therapeutic responses of DR and DME.

Keywords: diabetic retinopathy • clinical laboratory testing • aqueous 
© 2011, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
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