June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Targeting lipid-based nanoparticles promotes cell-specific gene delivery to eyes and improves vision in vivo
Author Affiliations & Notes
  • Yuhong Wang
    Ophthal/Dean McGee Eye Inst, Univ of Oklahoma Hlth Sci Ctr, Oklahoma City, OK
  • Ammaji Rajala
    Ophthal/Dean McGee Eye Inst, Univ of Oklahoma Hlth Sci Ctr, Oklahoma City, OK
  • Michelle Ranjo-Bishop
    Ophthal/Dean McGee Eye Inst, Univ of Oklahoma Hlth Sci Ctr, Oklahoma City, OK
  • Jian-xing Ma
    Medicine/Physiology, Univ of Oklahoma Hlth Sci Ctr, Oklahoma City, OK
  • Raju V S Rajala
    Ophthal/Dean McGee Eye Inst, Univ of Oklahoma Hlth Sci Ctr, Oklahoma City, OK
  • Footnotes
    Commercial Relationships Yuhong Wang, None; Ammaji Rajala, None; Michelle Ranjo-Bishop, None; Jian-xing Ma, None; Raju Rajala, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 5460. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Yuhong Wang, Ammaji Rajala, Michelle Ranjo-Bishop, Jian-xing Ma, Raju V S Rajala; Targeting lipid-based nanoparticles promotes cell-specific gene delivery to eyes and improves vision in vivo. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):5460.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: The application of viruses as carriers to deliver genes to eye tissue was successful, though unsafe. A safer, non-viral, biocompatible lipid-based nanoparticle to treat blinding eye diseases has never been tested in vivo. In this study, we created an artificial virus using a nanoparticle, liposome-protamine-DNA complex (LPD), to deliver a functional gene with sustained expression to eye tissues in a cell-specific manner for the treatment of eye disease.<br />

Methods: Liposomes were prepared by standard liposome technology and mixed with DNA, protamine, and nuclear localization signaling (NLS) peptide to generate lipid-protamine-DNA complex (LPD). We used LPD-complexed with GFP-reporter to monitor the in vivo gene transduction. Chicken Rpe65 cDNA or control plasmid DNA was complexed with LPD and injected into Rpe65 knockout mice. Cone function was assessed by electroretinography (ERG). Cone density was measured by lectin cytochemical analysis using peanut agglutinin (PNA). Localization of Rpe65 was examined by immunohistochemistry.<br />

Results: Our studies suggest that LPD can deliver GFP-reporter in vivo. Rpe65 protein is the key enzyme in regulating the availability of photochemicals. A deficiency of this gene results in blinding eye disease. Rpe65 mice do not show any ERG, due to the absence of chromophore regeneration. These mice also exhibit progressive cone cell loss by four weeks of age. LPD-Rpe65-gene delivery to Rpe65 knockout mice produced vision improvements. Our results indicated a complete loss of cones in Rpe65 knockout mice injected with control DNA, and preservation of cones in LPD-Rpe65 gene delivered Rpe65 knockout mice, comparable to wild-type controls. The immunohistochemistry results showed that Rpe65 expression was restricted to the retinal pigment epithelium layer in Rpe65-injected Rpe65 knockout mice.<br />

Conclusions: Our results demonstrate, for the first time, that LPD promotes efficient delivery in a cell-specific manner, and delivers a long-term expression of Rpe65 gene to mice lacking Rpe65 protein, leading to in vivo correction of blindness. Thus, LPD nanoparticles could provide a promising, efficient, non-viral method of gene delivery with clinical applications in eye disease treatment.<br />

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×