April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Ocular Delivery of Compacted DNA-Nanoparticles Does Not Elicit Toxicity in the Mouse Retina
Author Affiliations & Notes
  • X.-Q. Ding
    Cell Biology, Univ Oklahoma Health Sciences Center, Oklahoma City, Oklahoma
  • A. B. Quiambao
    Cell Biology, Univ Oklahoma Health Sciences Center, Oklahoma City, Oklahoma
  • J. B. Fitzgerald
    Cell Biology, Univ Oklahoma Health Sciences Center, Oklahoma City, Oklahoma
  • M. J. Cooper
    Copernicus Therapeutics, Inc., Cleveland, Ohio
  • S. Conley
    Cell Biology, Univ Oklahoma Health Sciences Center, Oklahoma City, Oklahoma
  • M. Naash
    Cell Biology, Univ Oklahoma Health Sciences Center, Oklahoma City, Oklahoma
  • Footnotes
    Commercial Relationships  X.-Q. Ding, None; A.B. Quiambao, None; J.B. Fitzgerald, None; M.J. Cooper, None; S. Conley, None; M. Naash, None.
  • Footnotes
    Support  NEI EY-017031 (XQD), NEI EY018656 (MIN), FFB, Inc. (MIN), and OCAST (MIN). Drs. Ashish Chintakuntlawar and James Chodosh’s support is acknowledged in facilitating these experiments.
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 1728. doi:
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    • Get Citation

      X.-Q. Ding, A. B. Quiambao, J. B. Fitzgerald, M. J. Cooper, S. Conley, M. Naash; Ocular Delivery of Compacted DNA-Nanoparticles Does Not Elicit Toxicity in the Mouse Retina. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1728.

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

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Abstract

Purpose: : Subretinal delivery of compacted DNA nanoparticles has been shown to direct effective transfection and expression in both photoreceptors and other retinal cells. This technology therefore provides a useful non-viral approach for gene transfer to treat inherited retinal diseases. Since one of the limitations to successful gene therapy has been vector-associated toxicity, this work evaluates the safety of compacted DNA nanoparticles following subretinal delivery.

Methods: : Polyethylene glycol-substituted lysine peptide-compacted nanoparticles containing an enhanced green fluorescent protein (EGFP) expression plasmid (pZEOGFP5.1) were sub-retinally injected in adult mice (1 µl at 0.3, 1.0 and 3.0 µg/µl). Mock and saline injections were performed as controls. Retinas were examined for transgene expression and for signs of inflammation at 1, 2, 4 and 7 days post-injection (PI). H&E staining on retinal sections was performed to detect infiltration of polymorphonuclear neutrophils (PMN) and other inflammatory cells. Immunohistochemical analyses were performed to detect expression of macrophage marker F4/80 and myeloid marker myeloperoxidase (MPO). Expression levels of chemokines KC, monocyte chemotactic protein-1 (MCP-1) and tumor necrosis factor alpha (TNF-) were evaluated by ELISA and by quantitative RT-PCR.

Results: : Subretinal delivery of the nanoparticles induced expression of EGFP in photoreceptors and retinal pigment epithelium cells. Neither infiltration of PMN or lymphocytes nor elevations of F4/80 or MPO were detected in retinas of mice that had been injected with nanoparticles. Levels of KC mRNA were increased 3-4 fold in eyes that had been injected with either nanoparticles or with saline at PI-1, but returned to control level at PI-2. No elevation of KC protein was observed in these mice. Levels of MCP-1 protein were increased 3-4 fold at PI-1 for both nanoparticle and saline injected eyes, but also returned to control levels at PI-2. Thus transient elevations of MCP-1 protein were related to the subretinal injection procedure. No elevations of TNF- mRNA or protein were detected.

Conclusions: : These investigations show no signs of local inflammatory responses or cellular toxicity following subretinal injection of compacted DNA nanoparticles, indicating that the retina may be a suitable target for future clinical nanoparticle-based interventions.

Keywords: retina • gene transfer/gene therapy • inflammation 
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