September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Adenosine deaminase-2-induced permeability increase in HRECs is suppressed by miR-146b-3p
Author Affiliations & Notes
  • Yara A Samra
    Biochemistry, Faculty of pharmacy, Mansoura University, Mansoura, Egypt
    The Culver Vision Discovery Institute, Ophthalmology, Medical College of Georgia, Augusta University, Augusta, Georgia, United States
  • Heba M Saleh
    Oral Biology and Anatomy, Dental College of Georgia, Augusta University, Augusta, Georgia, United States
  • Khaled Hussein
    Oral Biology and Anatomy, Dental College of Georgia, Augusta University, Augusta, Georgia, United States
  • Nehal M Elsherbiny
    Biochemistry, Faculty of pharmacy, Mansoura University, Mansoura, Egypt
    Oral Biology and Anatomy, Dental College of Georgia, Augusta University, Augusta, Georgia, United States
  • Ahmed Ibrahim
    Oral Biology and Anatomy, Dental College of Georgia, Augusta University, Augusta, Georgia, United States
    Biochemistry, Faculty of pharmacy, Mansoura University, Mansoura, Egypt
  • Sadanand Fulzele
    Orthopedics, Augusta University, Augusta, Georgia, United States
  • Mamdouh El-Shishtawy
    Biochemistry, Faculty of pharmacy, Mansoura University, Mansoura, Egypt
  • Laila A Eissa
    Biochemistry, Faculty of pharmacy, Mansoura University, Mansoura, Egypt
  • Mohamed Al-Sayed Al-Shabrawey
    Oral Biology and Anatomy, Dental College of Georgia, Augusta University, Augusta, Georgia, United States
    The Culver Vision Discovery Institute, Ophthalmology, Medical College of Georgia, Augusta University, Augusta, Georgia, United States
  • Gregory Ing Liou
    The Culver Vision Discovery Institute, Ophthalmology, Medical College of Georgia, Augusta University, Augusta, Georgia, United States
  • Footnotes
    Commercial Relationships   Yara Samra, None; Heba Saleh, None; Khaled Hussein, None; Nehal Elsherbiny, None; Ahmed Ibrahim, None; Sadanand Fulzele, None; Mamdouh El-Shishtawy, None; Laila Eissa, None; Mohamed Al-Shabrawey, None; Gregory Liou, None
  • Footnotes
    Support  Egyptian Cultural and Educational Bureau and Department of DefenseDM102155
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 5424. doi:
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      Yara A Samra, Heba M Saleh, Khaled Hussein, Nehal M Elsherbiny, Ahmed Ibrahim, Sadanand Fulzele, Mamdouh El-Shishtawy, Laila A Eissa, Mohamed Al-Sayed Al-Shabrawey, Gregory Ing Liou; Adenosine deaminase-2-induced permeability increase in HRECs is suppressed by miR-146b-3p. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5424.

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

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Abstract

Purpose : Diabetic retinopathy (DR), a neuroinflammatory disease, includes excessively increased cytokine release, microglia activation and blood–retinal barrier (BRB) breakdown. The molecular mechanism of neuroinflammation leading to BRB breakdown remains unclear. Adenosine deaminase-2 (ADA2), important for macrophage proliferation, is negatively regulated by miR-146b-3p. This is an initial study to investigate the role of miR-146b-3p and ADA2 in BRB breakdown.

Methods : Human monocytes U937 were differentiated to macrophages by phorbol 12-myristate 13-acetate (PMA 50ng/mL). Cells were then treated with amadori glycated albumin (AGA 500 µg/mL) for 12 hour. ADA2 activity in the macrophages conditioned medium (CM) and in the vitreous of diabetic and non-diabetic, non-inflammatory human donor eyes was colorimetrically measured. TNFα and IL-6 were measured in macrophages CM by enzyme-linked immunosorbent assay (ELISA). Human retinal endothelial cells (HRECs) were treated with AGA, CM of AGA-treated macrophages, and CM of AGA-treated and miR-146b-3p-transfected macrophages and BRB of HRECs was evaluated by electric cell-substrate impedance sensing (ECIS) that measures the changes in transcellular electrical resistance (TER). ZO-1 distribution in HRECs was examined by immunofluorescence.

Results : Our results illustrated that ADA2 activity was significantly increased in the diabetic humans as compared with the control (p<0.05). ADA2 activity, TNFα and IL-6 levels were increased significantly in the CM of AGA-treated macrophages as compared with the control (p<0.001). ADA2 activity, TNFα and IL-6 levels were significantly decreased by miR-146b-3p (p<0.05). TER of HRECs was significantly decreased by CM of AGA-treated macrophages as compared with AGA treatment (p<0.0001). This effect was significantly attenuated by miR-146b-3p, but not control, transfection (p<0.001). Lastly, ZO-1 pattern in HRECs was significantly disturbed by CM of AGA-treated macrophages as compared with AGA treatment. This effect was attenuated by miR-146b-3p transfection.

Conclusions : Our results suggest that ADA2 and miR-146b-3p contribute to BRB breakdown in DR. Thus, inhibition of ADA2 by miR-146b-3p is a potential therapeutic strategy to preserve retinal endothelial cell barrier function in DR.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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