July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Extracellular vesicles from endothelial colony forming cells as paracrine mediators of neurovasculotrophic repair of the retina
Author Affiliations & Notes
  • Kyle V Marra
    Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, United States
    Department of Bioengineering, University of California, San Diego, La Jolla, California, United States
  • Susumu Sakimoto
    Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, United States
  • Salome Murinello
    Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, United States
  • Edith Aguilar
    Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, United States
  • Felicitas Bucher
    Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, United States
  • Martin Friedlander
    Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, United States
  • Footnotes
    Commercial Relationships   Kyle Marra, None; Susumu Sakimoto, None; Salome Murinello, None; Edith Aguilar, None; Felicitas Bucher, None; Martin Friedlander, None
  • Footnotes
    Support  NH Grant EY11254, The Lowy Medical Research Institute; Scripps Clinical Medical Group Research Fund; University of California, San Diego Medical Scientist Training Program T32 GM007198-40
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 5489. doi:
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    • Get Citation

      Kyle V Marra, Susumu Sakimoto, Salome Murinello, Edith Aguilar, Felicitas Bucher, Martin Friedlander; Extracellular vesicles from endothelial colony forming cells as paracrine mediators of neurovasculotrophic repair of the retina. Invest. Ophthalmol. Vis. Sci. 2018;59(9):5489.

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

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Abstract

Purpose : Endothelial colony forming cells (ECFCs) are a subset of endothelial progenitor cells that promote physiologic vascular growth and stability via a paracrine action. Recent studies have shown that microRNA-containing extracellular vesicles (EVs) shed from ECFCs reduced the area of vaso-obliteration and targeted genes associated with angiogenesis in the murine oxygen-induced retinopathy (OIR) model. In this study we investigate an EV-mediated paracrine mechanism underlying the observations that a subset of ECFCs characterized by high expression of the surface marker CD44 (CD44hi) exhibit an enhanced neuro- and vasculotrophic paracrine effect in rodent models of ischemic retinopathy, compared to cells with low expression of CD44 (CD44low).

Methods : To obtain pure samples of EVs, ECFC conditioned media was processed by a combination of differential ultracentrifugation, size exclusion chromatography, and ultrafiltration. OIR was induced in C57BL/6J mice via 75% oxygen exposure from p7-p12, after which mice were treated with EVs from CD44hi or CD44low ECFCs. EV samples were then characterized via metabolomic and proteomic analysis as well as microRNA sequencing. Intravesicular cargo significantly upregulated in EVs from CD44hi ECFCs were selected for in vitro functional assays of angiogenesis.

Results : In OIR, injections of EVs derived from CD44hi ECFCs significantly reduced the area of retinal neovascularization (NV) and vaso-obliteration (VO) in comparison to injections of EVs from CD44low ECFCs (n=25; NV: 1.65% vs. 9.07%, p<0.0001; VO: 6.43% vs. 16.52%, p<0.0001). Size-exclusion chromatography of media conditioned by ECFCs allowed for the isolation of EV samples free of soluble protein, which facilitated the downstream characterization of intravesicular bioactive cargo. We use metabolomic, proteomic, and sequencing analyses to identify agents that may mediate the therapeutic effect of CD44hi EVs.

Conclusions : By comparing the bioactive cargo of EVs from therapeutic CD44hi ECFCs in comparison to ineffective CD44low EVs, our -omics based approach may be used to identify novel therapeutic microRNAs, proteins, and small molecules that promote CD44hi EV-mediated rescue effect in the OIR model. EVs may be a highly effective drug delivery vehicle that circumvents the risks of tumorigenesis associated with cell-based therapeutics.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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