June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Transduction of Mouse Retina by Intravitreal Delivery Using Chimeric Capsids with Altered Glycan Interactions
Author Affiliations & Notes
  • Kenton Woodard
    Gene Therapy Center, University of North Carolina, Chapel Hill, NC
    Neurobiology Curriculum, University of North Carolina, Chapel Hill, NC
  • Katharine J Liang
    Gene Therapy Center, University of North Carolina, Chapel Hill, NC
    Neurobiology Curriculum, University of North Carolina, Chapel Hill, NC
  • Thomas Lentz
    Gene Therapy Center, University of North Carolina, Chapel Hill, NC
  • William Bennett
    Gene Therapy Center, University of North Carolina, Chapel Hill, NC
  • Aravind Asokan
    Gene Therapy Center, University of North Carolina, Chapel Hill, NC
    Genetics, University of North Carolina, Chapel Hill, NC
  • Richard Samulski
    Gene Therapy Center, University of North Carolina, Chapel Hill, NC
    Pharmacology, University of North Carolina, Chapel Hill, NC
  • Footnotes
    Commercial Relationships Kenton Woodard, None; Katharine Liang, None; Thomas Lentz, None; William Bennett, None; Aravind Asokan, None; Richard Samulski, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 252. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Kenton Woodard, Katharine J Liang, Thomas Lentz, William Bennett, Aravind Asokan, Richard Samulski; Transduction of Mouse Retina by Intravitreal Delivery Using Chimeric Capsids with Altered Glycan Interactions. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):252.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: Intravitreal (IVit) delivery of recombinant adeno-associated virus (rAAV) has advantages over subretinal delivery, which is the clinic standard. This study sought to correlate capsid-receptor interaction to transduction using a panel of variants with modulated glycan interactions.

Methods: Serotypes with altered heparan sulfate (HS) binding were evaluated using the GFP reporter gene. Dual glycan interaction of HS and galactose binding was evaluated using the chimera rAAV2G9. Fundus imaging documented fluorescence onset and pattern. Eight weeks post-injection, histological analyses were conducted to identify cell tropism and retinal transduction. Fluorescence in situ hybridization (FISH) was used to detect viral genome location and immunohistochemistry was used to detect viral transduction.

Results: Mutations in HS binding residues of rAAV2 resulted in a loss of transduction of the inner retina. The addition of HS binding to rAAV1 showed a trend towards increased fluorescence over rAAV1, but tropism was unaltered and similar to published results with rAAV6. rAAV2G9 had similar GFP intensity to rAAV2; however, histology revealed an increase in Muller glia tropism with the chimera. This tropism difference was not due to damage to the retina or reactivation of glial cells. FISH results suggest viral genomes are present in the retina even in the absence of GFP fluorescence.

Conclusions: HS binding is required for IVit transduction of rAAV2, and may enhance the transduction of other serotypes. Galactose binding on rAAV2 alters infectivity towards Muller glia cells through receptor interactions with the capsid. The absence of rAAV9 expression, but the presence of genomes in the retina, indicate additional cellular barriers which restrict expression. The development of rAAV receptor heatmaps of the retina will improve the rational engineering of vectors for targeted, and more efficient, retinal gene transfer.

×
×

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.

×