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Safety and angiogenic effects of systemic gene delivery of a modified erythropoietin

Gene Therapy volume 22, pages 365–373 (2015)Cite this article

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Abstract

Erythropoietin (EPO) is critical for red blood cell production and is also an effective neuroprotective agent. However, it may contribute to pathological angiogenesis. Here we investigate the angiogenic potential of EPO and a mutant form with attenuated erythropoietic activity, EPO-R76E, on primary human retinal microvascular endothelial cells (HRMECs) and in the adult retina. Assays of death, proliferation and tube formation were performed on HRMECs exposed to EPO, EPO-R76E or media alone. Postnatal day-9 wild-type mice were injected intramuscularly with adeno-associated virus vectors expressing either enhanced green fluorescent protein or EpoR76E. At 3 months, levels of EPO-R76E in the eye were quantified, and the health of the retinal vasculature was assessed by fluorescein angiography and isolectin immunolabeling. Immunohistochemistry, histology and electroretinogram (ERG) assessments were performed as measures of retinal health. Neither EPO nor EPO-R76E induced proliferation or tube formation in HRMECs under the conditions used. EPO-R76E decreased HRMEC death in a dose-dependent manner. Long-term systemic gene delivery of EPO-R76E was safe in terms of retinal vasculature, histology and the ERG in vivo. Our results show that EPO-R76E can block HRMEC death, consistent with its role in erythropoiesis and neuroprotection. In addition, long-term gene delivery of EPO-R76E is safe in the adult retina.

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Acknowledgements

We thank Megan Capozzi in Dr John Penn’s laboratory for assistance with the tube-formation assay, Dr Ashwath Jayagopal for assistance with fluorescein angiography and Kristi Wynn for technical assistance. Stable cell lines were generated and EPO and EPO-R76E were quantified by the VAPR, which is supported by the Vanderbilt Institute of Chemical Biology and the Vanderbilt Ingram Cancer Center (P30 CA68485). This work was supported by grants from Research to Prevent Blindness Career Development Award; Research to Prevent Blindness Unrestricted Funds (P. Sternberg); CTSA award UL1TR000445 from the National Center for Advancing Translational Sciences; Department of Defense W81XW-10-1-0528; P30-EY008126; NEI EY022349; and Fight for Sight.

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  1. Department of Ophthalmology and Visual Sciences, Vanderbilt Eye Institute, Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, USA

    A M de Lucas Cerrillo, W S Bond & T S Rex

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  1. A M de Lucas Cerrillo
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Correspondence to T S Rex.

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Competing interests

TSR is a co-inventor on a patent application (US No. 13/979,451) regarding neuroprotective action of EPO-R76E. No licensing has occurred. The other authors declare no conflict of interest.

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The views, opinions and/or findings contained in this research paper are those of the authors and do not necessarily reflect the views of the Department of Defense and should not be construed as an official DoD/Army position, policy or decision unless so designated by other documentation. No official endorsement should be made.

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de Lucas Cerrillo, A., Bond, W. & Rex, T. Safety and angiogenic effects of systemic gene delivery of a modified erythropoietin. Gene Ther 22, 365–373 (2015). https://doi.org/10.1038/gt.2015.12

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