June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Crosstalk between RPE and choroid endothelium regulates RPE tight junctions
Author Affiliations & Notes
  • Ignacio Benedicto
    Ophtalmology, Weill Cornell Medical College, New York, NY
    Margaret Dyson Vision Research Institute, Weill Cornell Medical College, New York, NY
  • Guillermo Lehmann-Mantaras
    Ophtalmology, Weill Cornell Medical College, New York, NY
    Margaret Dyson Vision Research Institute, Weill Cornell Medical College, New York, NY
  • Michael Ginsberg
    Angiocrine Bioscience, New York, NY
  • Daniel J. Nolan
    Angiocrine Bioscience, New York, NY
  • Olivier Elemento
    Physiology and Biophysics, Weill Cornell Medical College, New York, NY
  • Shahin Rafii
    Angiocrine Bioscience, New York, NY
    Genetic Medicine, Weill Cornell Medical College, New York, NY
  • Enrique Rodriguez-Boulan
    Ophtalmology, Weill Cornell Medical College, New York, NY
    Margaret Dyson Vision Research Institute, Weill Cornell Medical College, New York, NY
  • Footnotes
    Commercial Relationships Ignacio Benedicto, None; Guillermo Lehmann-Mantaras, None; Michael Ginsberg, Angiocrine Bioscience (E), Angiocrine Bioscience (I); Daniel Nolan, Angiocrine Bioscience (E), Angiocrine Bioscience (I); Olivier Elemento, None; Shahin Rafii, Angiocrine Bioscience (I); Enrique Rodriguez-Boulan, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 41. doi:
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    • Get Citation

      Ignacio Benedicto, Guillermo Lehmann-Mantaras, Michael Ginsberg, Daniel J. Nolan, Olivier Elemento, Shahin Rafii, Enrique Rodriguez-Boulan; Crosstalk between RPE and choroid endothelium regulates RPE tight junctions. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):41.

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

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Abstract

Purpose: The retina is irrigated by two blood circulations, the choroid and the retinal blood vessels. The retinal pigment epithelium (RPE) performs a key role as a perm-selective barrier between the neural retina and the fenestrated choroid capillaries. RPE tight junctions (TJs) are a key component of this blood-retinal barrier. Recent studies have demonstrated that endothelial cells (ECs) display organ-specific transcriptomes and play key instructive roles in the differentiation and maintenance of parenchymal cells. Here, we isolated mouse choroid ECs (ChECs) and retinal ECs to high purity, analyzed their transcriptome relative to each other and to other body ECs and studied the regulation of RPE TJs by ChECs.

Methods: We performed co-culture experiments on Transwell inserts using human umbilical vein ECs expressing the adenoviral protein E4 (VeraVecs), which are able to survive in the absence of endothelial factors or serum, and human fetal RPE (hfRPE). The effect of VeraVecs on hfRPE transepithelial electrical resistance (TER) was assessed. Expression levels of occludin and different claudins in hfRPE were analyzed by real time PCR. The effect of the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor AG1478 on hfRPE TER and claudin levels was also evaluated. We isolated mouse ChECs by in vivo EC labeling and flow cytometry cell sorting. ChEC conditioned medium was used in hfRPE wound healing assays. We performed RNAseq analysis of mouse native ECs extracted from choroid, neural retina, heart, liver and lung.

Results: VeraVec and ChEC-conditioned media specifically induced an increase in hfRPE TER during both de novo TJ formation and TJ recovery after disruption by TNFα treatment, and a decrease in hfRPE claudin-2 mRNA levels. Bioinformatic analyses showed that ChECs presented a significantly high expression of a set of genes involved in development and wound healing, including the EGFR ligand HBEGF. Recombinant HBEGF increased hfRPE TER, and EGFR tyrosine kinase inhibition reduced EC-mediated changes in hfRPE TER and claudin-2 levels. Wound healing assays showed that ChEC conditioned media improved hfRPE wound closure.

Conclusions: Our results suggest a role of ChECs in the formation and maintenance of the outer blood-retinal barrier by modulating RPE TJ function, and in repair processes after RPE injury. Thus, ChECs emerge as a potential new therapeutic tool to treat RPE-related vision pathologies.

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