June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Overexpression of ELOVL4 stabilizes tight junctions and prevents VEGF-induced vascular permeability in diabetic retina and retinal cells.
Author Affiliations & Notes
  • Nermin Kady
    Physiology, Michigan State university, East Lansing, Michigan, United States
  • Xuwen Liu
    Ophthalmology and Visual Sciences, University of Michigan -Kellogg eye center, Ann arbor , Michigan, United States
  • Todd Lydic
    Physiology, Michigan State university, East Lansing, Michigan, United States
  • Segey Seregin
    University of Michigan, Ann arbor, Michigan, United States
  • Andrea Amalfitano
    Microbiology and Molecular Genetics, Michigan State University, East Lansing, Michigan, United States
  • Vince A Chiodo
    University of Florida , Gainesville, Florida, United States
  • Sanford L Boye
    University of Florida , Gainesville, Florida, United States
  • William Hauswirth
    University of Florida , Gainesville, Florida, United States
  • Maria B Grant
    Opthalmology, University of Indiana, Bloomington, Indiana, United States
  • David Antonetti
    Ophthalmology and Visual Sciences, University of Michigan -Kellogg eye center, Ann arbor , Michigan, United States
  • Julia V Busik
    Physiology, Michigan State university, East Lansing, Michigan, United States
  • Footnotes
    Commercial Relationships   Nermin Kady, None; Xuwen Liu, None; Todd Lydic, None; Segey Seregin, None; Andrea Amalfitano, None; Vince Chiodo, None; Sanford Boye, None; William Hauswirth, AGTC (I), Bionic Sight (I); Maria Grant, None; David Antonetti, None; Julia Busik, None
  • Footnotes
    Support  RO1EY016077 NIH/NEI, R01EY025383 NIH/NEI, AgBioResearch grant MICL02163, NIH EY012021 and Jules and Doris Stein Professorship,NIH P30EY021721 (WWH), Research to Prevent Blindness (Univ. of Florida)
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 4034. doi:
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      Nermin Kady, Xuwen Liu, Todd Lydic, Segey Seregin, Andrea Amalfitano, Vince A Chiodo, Sanford L Boye, William Hauswirth, Maria B Grant, David Antonetti, Julia V Busik; Overexpression of ELOVL4 stabilizes tight junctions and prevents VEGF-induced vascular permeability in diabetic retina and retinal cells.. Invest. Ophthalmol. Vis. Sci. 2017;58(8):4034.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : We have previously demonstrated that the expression level of elongation of very long chain fatty acids –like 4(ELOVL4) is significantly reduced in the diabetic retina leading to decreased levels of very long chain (VLC ≥26:2) fatty acids. VLC fatty acids produced by ELOVL4 incorporate into VLC ceramides that are known to play an important role in skin permeability barrier maintenance. We hypothesized that, similar to skin, VLC ceramides could play a beneficial role in maintenance of the blood-retinal barrier and reduction of ELOVL4 in diabetes could lead to increased retinal vascular permeability through the decrease in VLC ceramides.

Methods : Human ELOVL4 was overexpressed in bovine retinal endothelial cells (BRECs) by an E1- and E3-deleted adenoviral vector under the control of CMV promoter. Permeability was determined in confluent monolayers by RITC dextran. Tight junction proteins were assayed by Western blot and immunostaining. Ceramide colocalization with tight junction complex was determined by immunogold electron microscopy. Tight junction isolates were analyzed by tandem mass spectrometry. Streptozotocin (STZ) mice received intravitreal injection of hELOVL4 packaged in adeno-associated virus type 2 containing four capsid Y-F mutations (AAV2 mut quad) under the control of a CMV-chicken β-actin (smCBA) promoter or empty AAV2 mut quad as a negative control. Retinal vascular permeability was assessed by measuring extravasation of FITC-albumin after 8 weeks of diabetes.

Results : Overexpression of hELOVL4 in BRECs significantly decreased basal permeability by 46% (n=3, P<0.01), as well as VEGF and IL-1β-induced permeability by 47.7% and 36.2%; respectively (n= 3-6, p<0.001, P<0.01; respectively). Importantly, hELOVL4 overexpression prevented VEGF-induced decrease in occludin expression and border staining of tight junction proteins; ZO-1 and claudin-5. Ceramides were found to colocalize with tight junction complexes. Lipidome analysis of tight junction isolates revealed the presence of VLC ceramides. Intravitreal delivery of hELOVL4-AAV2 in STZ mice reduced diabetes-induced increase in vascular permeability by 75% after 8 weeks of diabetes (n= 8-12 per group, p<0.01).

Conclusions : Normalization of retinal ELOVL4 expression could prevent blood-retina barrier dysregulation in DR through increase in VLC ceramides and stabilization of tight junctions.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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