Adenosine deaminase-2–induced hyperpermeability in human retinal vascular endothelial cells is suppressed by microRNA-146b-3p

Yara A. Samra, Heba Mohamed Saleh, Khaled A. Hussein, Nehal M. Elsherbiny, Ahmed Salah Ibrahim, Khaled Elmasry, Sadanand T Fulzele, Mamdouh M. El-Shishtawy, Laila A. Eissa, Mohamed Al-Shabrawey, Gregory I Liou

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

PURPOSE. We recently demonstrated that adenosine deaminase-2 (ADA2) contributes to diabetic retinopathy (DR) via up-regulating the production of inflammatory cytokines in macrophages. Also, microRNA (miR)-146b-3p has the ability to inhibit ADA2. The goal of this study was to investigate the potential role of ADA2 and therapeutic benefit of miR-146b-3p in retinal inflammation and endothelial barrier dysfunction during diabetes. METHODS. Adenosine deaminase-2 activity was determined by colorimetric method in diabetic human vitreous. Human monocyte cell line U937 was differentiated into macrophages and then treated with amadori glycated albumin (AGA), and conditioned medium (CM) was used to assess the changes in ADA2 activity and TNF-α and IL-6 levels by ELISA. Also, macrophages were transfected with miR-146b-3p before treatment with AGA. Permeability of human retinal endothelial cells (hRECs) was assessed by electric cell-substrate impedance sensing (ECIS) after treatment with macrophage CM. Zonula occludens (ZO)-1 was examined by immuno-fluorescence in hRECs. Leukocyte adhesion was assessed in hRECs by measuring myeloperoxidase (MPO) activity and intercellular adhesion molecule-1 (ICAM-1) expression. RESULTS. Adenosine deaminase-2 activity was significantly increased in diabetic human vitreous. ADA2 activity and TNF-α and IL-6 levels were significantly increased in human macrophages by AGA treatment. Amadori glycated albumin–treated macrophage CM significantly increased hREC permeability, disrupted ZO-1 pattern, and increased leukocyte adhesion to hRECs through up-regulating ICAM-1. All these changes were reversed by miR-146b-3p. CONCLUSIONS. Adenosine deaminase-2 is implicated in breakdown of the blood–retinal barrier (BRB) in DR through macrophages-derived cytokines. Therefore, inhibition of ADA2 by miR-146b-3p might be a useful tool to preserve BRB function in DR.

Original languageEnglish (US)
Pages (from-to)933-943
Number of pages11
JournalInvestigative Ophthalmology and Visual Science
Volume58
Issue number2
DOIs
StatePublished - Feb 1 2017

Fingerprint

Retinal Vessels
Adenosine Deaminase
MicroRNAs
Endothelial Cells
Macrophages
Diabetic Retinopathy
Conditioned Culture Medium
Tight Junctions
Intercellular Adhesion Molecule-1
Permeability
Interleukin-6
Leukocytes
Cytokines
Therapeutics
Electric Impedance
Peroxidase
Monocytes
Fluorescence
Enzyme-Linked Immunosorbent Assay
Inflammation

Keywords

  • Adenosine deaminase-2
  • BRB
  • Diabetic retinopathy
  • IL-6
  • Leukocyte adhesion
  • MiR-146b-3p
  • TNF-α
  • ZO-1

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Adenosine deaminase-2–induced hyperpermeability in human retinal vascular endothelial cells is suppressed by microRNA-146b-3p. / Samra, Yara A.; Saleh, Heba Mohamed; Hussein, Khaled A.; Elsherbiny, Nehal M.; Ibrahim, Ahmed Salah; Elmasry, Khaled; Fulzele, Sadanand T; El-Shishtawy, Mamdouh M.; Eissa, Laila A.; Al-Shabrawey, Mohamed; Liou, Gregory I.

In: Investigative Ophthalmology and Visual Science, Vol. 58, No. 2, 01.02.2017, p. 933-943.

Research output: Contribution to journalArticle

Samra, Yara A. ; Saleh, Heba Mohamed ; Hussein, Khaled A. ; Elsherbiny, Nehal M. ; Ibrahim, Ahmed Salah ; Elmasry, Khaled ; Fulzele, Sadanand T ; El-Shishtawy, Mamdouh M. ; Eissa, Laila A. ; Al-Shabrawey, Mohamed ; Liou, Gregory I. / Adenosine deaminase-2–induced hyperpermeability in human retinal vascular endothelial cells is suppressed by microRNA-146b-3p. In: Investigative Ophthalmology and Visual Science. 2017 ; Vol. 58, No. 2. pp. 933-943.
@article{765bbcfae81c42a9bdf77f40cd03fc25,
title = "Adenosine deaminase-2–induced hyperpermeability in human retinal vascular endothelial cells is suppressed by microRNA-146b-3p",
abstract = "PURPOSE. We recently demonstrated that adenosine deaminase-2 (ADA2) contributes to diabetic retinopathy (DR) via up-regulating the production of inflammatory cytokines in macrophages. Also, microRNA (miR)-146b-3p has the ability to inhibit ADA2. The goal of this study was to investigate the potential role of ADA2 and therapeutic benefit of miR-146b-3p in retinal inflammation and endothelial barrier dysfunction during diabetes. METHODS. Adenosine deaminase-2 activity was determined by colorimetric method in diabetic human vitreous. Human monocyte cell line U937 was differentiated into macrophages and then treated with amadori glycated albumin (AGA), and conditioned medium (CM) was used to assess the changes in ADA2 activity and TNF-α and IL-6 levels by ELISA. Also, macrophages were transfected with miR-146b-3p before treatment with AGA. Permeability of human retinal endothelial cells (hRECs) was assessed by electric cell-substrate impedance sensing (ECIS) after treatment with macrophage CM. Zonula occludens (ZO)-1 was examined by immuno-fluorescence in hRECs. Leukocyte adhesion was assessed in hRECs by measuring myeloperoxidase (MPO) activity and intercellular adhesion molecule-1 (ICAM-1) expression. RESULTS. Adenosine deaminase-2 activity was significantly increased in diabetic human vitreous. ADA2 activity and TNF-α and IL-6 levels were significantly increased in human macrophages by AGA treatment. Amadori glycated albumin–treated macrophage CM significantly increased hREC permeability, disrupted ZO-1 pattern, and increased leukocyte adhesion to hRECs through up-regulating ICAM-1. All these changes were reversed by miR-146b-3p. CONCLUSIONS. Adenosine deaminase-2 is implicated in breakdown of the blood–retinal barrier (BRB) in DR through macrophages-derived cytokines. Therefore, inhibition of ADA2 by miR-146b-3p might be a useful tool to preserve BRB function in DR.",
keywords = "Adenosine deaminase-2, BRB, Diabetic retinopathy, IL-6, Leukocyte adhesion, MiR-146b-3p, TNF-α, ZO-1",
author = "Samra, {Yara A.} and Saleh, {Heba Mohamed} and Hussein, {Khaled A.} and Elsherbiny, {Nehal M.} and Ibrahim, {Ahmed Salah} and Khaled Elmasry and Fulzele, {Sadanand T} and El-Shishtawy, {Mamdouh M.} and Eissa, {Laila A.} and Mohamed Al-Shabrawey and Liou, {Gregory I}",
year = "2017",
month = "2",
day = "1",
doi = "10.1167/iovs.16-19782",
language = "English (US)",
volume = "58",
pages = "933--943",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology Inc.",
number = "2",

}

TY - JOUR

T1 - Adenosine deaminase-2–induced hyperpermeability in human retinal vascular endothelial cells is suppressed by microRNA-146b-3p

AU - Samra, Yara A.

AU - Saleh, Heba Mohamed

AU - Hussein, Khaled A.

AU - Elsherbiny, Nehal M.

AU - Ibrahim, Ahmed Salah

AU - Elmasry, Khaled

AU - Fulzele, Sadanand T

AU - El-Shishtawy, Mamdouh M.

AU - Eissa, Laila A.

AU - Al-Shabrawey, Mohamed

AU - Liou, Gregory I

PY - 2017/2/1

Y1 - 2017/2/1

N2 - PURPOSE. We recently demonstrated that adenosine deaminase-2 (ADA2) contributes to diabetic retinopathy (DR) via up-regulating the production of inflammatory cytokines in macrophages. Also, microRNA (miR)-146b-3p has the ability to inhibit ADA2. The goal of this study was to investigate the potential role of ADA2 and therapeutic benefit of miR-146b-3p in retinal inflammation and endothelial barrier dysfunction during diabetes. METHODS. Adenosine deaminase-2 activity was determined by colorimetric method in diabetic human vitreous. Human monocyte cell line U937 was differentiated into macrophages and then treated with amadori glycated albumin (AGA), and conditioned medium (CM) was used to assess the changes in ADA2 activity and TNF-α and IL-6 levels by ELISA. Also, macrophages were transfected with miR-146b-3p before treatment with AGA. Permeability of human retinal endothelial cells (hRECs) was assessed by electric cell-substrate impedance sensing (ECIS) after treatment with macrophage CM. Zonula occludens (ZO)-1 was examined by immuno-fluorescence in hRECs. Leukocyte adhesion was assessed in hRECs by measuring myeloperoxidase (MPO) activity and intercellular adhesion molecule-1 (ICAM-1) expression. RESULTS. Adenosine deaminase-2 activity was significantly increased in diabetic human vitreous. ADA2 activity and TNF-α and IL-6 levels were significantly increased in human macrophages by AGA treatment. Amadori glycated albumin–treated macrophage CM significantly increased hREC permeability, disrupted ZO-1 pattern, and increased leukocyte adhesion to hRECs through up-regulating ICAM-1. All these changes were reversed by miR-146b-3p. CONCLUSIONS. Adenosine deaminase-2 is implicated in breakdown of the blood–retinal barrier (BRB) in DR through macrophages-derived cytokines. Therefore, inhibition of ADA2 by miR-146b-3p might be a useful tool to preserve BRB function in DR.

AB - PURPOSE. We recently demonstrated that adenosine deaminase-2 (ADA2) contributes to diabetic retinopathy (DR) via up-regulating the production of inflammatory cytokines in macrophages. Also, microRNA (miR)-146b-3p has the ability to inhibit ADA2. The goal of this study was to investigate the potential role of ADA2 and therapeutic benefit of miR-146b-3p in retinal inflammation and endothelial barrier dysfunction during diabetes. METHODS. Adenosine deaminase-2 activity was determined by colorimetric method in diabetic human vitreous. Human monocyte cell line U937 was differentiated into macrophages and then treated with amadori glycated albumin (AGA), and conditioned medium (CM) was used to assess the changes in ADA2 activity and TNF-α and IL-6 levels by ELISA. Also, macrophages were transfected with miR-146b-3p before treatment with AGA. Permeability of human retinal endothelial cells (hRECs) was assessed by electric cell-substrate impedance sensing (ECIS) after treatment with macrophage CM. Zonula occludens (ZO)-1 was examined by immuno-fluorescence in hRECs. Leukocyte adhesion was assessed in hRECs by measuring myeloperoxidase (MPO) activity and intercellular adhesion molecule-1 (ICAM-1) expression. RESULTS. Adenosine deaminase-2 activity was significantly increased in diabetic human vitreous. ADA2 activity and TNF-α and IL-6 levels were significantly increased in human macrophages by AGA treatment. Amadori glycated albumin–treated macrophage CM significantly increased hREC permeability, disrupted ZO-1 pattern, and increased leukocyte adhesion to hRECs through up-regulating ICAM-1. All these changes were reversed by miR-146b-3p. CONCLUSIONS. Adenosine deaminase-2 is implicated in breakdown of the blood–retinal barrier (BRB) in DR through macrophages-derived cytokines. Therefore, inhibition of ADA2 by miR-146b-3p might be a useful tool to preserve BRB function in DR.

KW - Adenosine deaminase-2

KW - BRB

KW - Diabetic retinopathy

KW - IL-6

KW - Leukocyte adhesion

KW - MiR-146b-3p

KW - TNF-α

KW - ZO-1

UR - http://www.scopus.com/inward/record.url?scp=85012034065&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85012034065&partnerID=8YFLogxK

U2 - 10.1167/iovs.16-19782

DO - 10.1167/iovs.16-19782

M3 - Article

C2 - 28170537

AN - SCOPUS:85012034065

VL - 58

SP - 933

EP - 943

JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

IS - 2

ER -