April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Primary RPE Express High Levels Of Epithelial Membrane Protein 2 (EMP2)-Implications For PVR Progression
Author Affiliations & Notes
  • Lynn K. Gordon
    Jules Stein Eye Inst,
    University of California, Los Angeles, Los Angeles, California
  • Shawn A. Morales
    Jules Stein Eye Inst,
    University of California, Los Angeles, Los Angeles, California
  • David G. Telander
    Ophthalmology, University of California, Davis, Sacramento, California
  • David R. Hinton
    Pathology, Keck School of Medicine University of Southern California, Los Angeles, California
  • Christine Spree
    Pathology, Keck School of Medicine University of Southern California, Los Angeles, California
  • Jonathan Braun
    Pathology and Laboratory Medicine,
    University of California, Los Angeles, Los Angeles, California
  • Footnotes
    Commercial Relationships  Lynn K. Gordon, None; Shawn A. Morales, None; David G. Telander, None; David R. Hinton, None; Christine Spree, None; Jonathan Braun, None
  • Footnotes
    Support  NIH Grant EY019909
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 6591. doi:
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      Lynn K. Gordon, Shawn A. Morales, David G. Telander, David R. Hinton, Christine Spree, Jonathan Braun; Primary RPE Express High Levels Of Epithelial Membrane Protein 2 (EMP2)-Implications For PVR Progression. Invest. Ophthalmol. Vis. Sci. 2011;52(14):6591.

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

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Abstract

Purpose: : The retinal pigment epithelium (RPE) plays an important role in proliferative vitreoretinopathy (PVR), an aberrant wound-healing process caused in part by RPE cell migration, membrane formation, and contractile cellular forces leading to tractional retinal detachment. Using an in vitro model of PVR, collagen gel contraction, we have previously shown that EMP2 regulates collagen gel contraction by facilitating FAK recruitment and activation in ARPE-19 cells. The purpose of this study is to determine if EMP2 is expressed in primary RPE cells and whether blockade of EMP2 could control contraction of collagen gels, an in vitro model for mechanisms involved in PVR.

Methods: : Primary human RPE cells were cultured according to published methods and used in these studies. EMP2 and p-FAK expression was assessed in ARPE-19 and primary RPE cells by Western blot. Collagen gel contraction was measured using an in vitro assay. Cell viability was calculated by Trypan blue exclusion. EMP2 localization was determined by confocal microscopy using polarized primary RPE cells.

Results: : Primary RPE cells express high levels of EMP2 and p-FAK. Expression is increased 88% and 125% respectively for EMP2 and p-FAK when compared to ARPE-19 cells (P<0.05). Expression of EMP2 is largely localized to the apical cell surface in the polarized epithelium. Primary RPE cells are able to contract a collagen gel and, concordant with the high levels of EMP2, their contractile capacity is increased 55% as compared to ARPE-19 cells (P<0.05). Gel contraction by the primary RPE cells is significantly reduced following reduction of EMP2 using an anti-EMP2 diabody.

Conclusions: : Human primary RPE cells express high levels of EMP2 and p-FAK and demonstrate a robust contractile capacity. The connection between these three factors as seen in ARPE-19 cells is preserved in primary RPE cells. Targeting EMP2 with and anti-EMP2 antibody reduces contraction and may serve as a novel therapeutic target in prevention of PVR.

Keywords: retinal pigment epithelium • proliferative vitreoretinopathy • retinal detachment 
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