May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Preclinical Evaluation of rAAV.sFlt-1 in Monkeys: Toxicity and Biodistribution
Author Affiliations & Notes
  • E. P. Rakoczy
    Centre for Ophthalmology & Visual Sciences, University of Western Australia, Perth, Australia
  • C.-M. Lai
    Centre for Ophthalmology & Visual Sciences, University of Western Australia, Perth, Australia
  • S. Lee
    Singapore Eye Research Institute, Singapore, Singapore
  • I. Yeo
    Singapore Eye Research Institute, Singapore, Singapore
  • R. Himbeck
    Molecular Ophthalmology, Lions Eye Institute, Perth, Australia
  • M. Escourt
    Centre for Ophthalmology & Visual Sciences, University of Western Australia, Perth, Australia
  • L. Chi
    Singapore Eye Research Institute, Singapore, Singapore
  • C. Ang
    Singapore Eye Research Institute, Singapore, Singapore
  • M. Degli-Espoti
    Centre for Ophthalmology & Visual Sciences, University of Western Australia, Perth, Australia
  • Footnotes
    Commercial Relationships  E.P. Rakoczy, None; C. Lai, None; S. Lee, None; I. Yeo, None; R. Himbeck, None; M. Escourt, None; L. Chi, None; C. Ang, None; M. Degli-Espoti, None.
  • Footnotes
    Support  NHMRC, JDRF
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 1132. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      E. P. Rakoczy, C.-M. Lai, S. Lee, I. Yeo, R. Himbeck, M. Escourt, L. Chi, C. Ang, M. Degli-Espoti; Preclinical Evaluation of rAAV.sFlt-1 in Monkeys: Toxicity and Biodistribution. Invest. Ophthalmol. Vis. Sci. 2008;49(13):1132. doi: https://doi.org/.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : To conduct toxicity and biodistribution studies in order to fulfil the requirements of the Australian Therapeutic Goods Administration (equivalent to FDA) authority to commence Phase I/II human trials.

Methods: : All animal experimentation was conducted in compliance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research. Monkeys (M. fascicularis) were subretinally injected with rAAV.sFlt-1 (n=5), rAAV.gfp (n=2) or sFLT protein (n=1). Animals were subjected to regular clinical and ophthalmic examinations which included slit lamp examination, electroretinography and fundus photography. Peripherial blood was collected at different time points. At the time of euthanasia, tissue samples were collected. Real-time PCR, ELISA and flow cytometry were used to establish recombinant virus and protein dissemination and immune response.

Results: : All monkeys were healthy throughout the course of the study. Fundoscopic examination revealed a normal fundus and clear vitreous. Slit lamp examination did not detect any abnormality in the anterior segment. There were no differences in a-wave and b-wave amplitudes taken before and after injection. Real-time PCR did not detect the presence of the AAV vector in the tissues sampled. ELISA did not detect any AAV2 capsid or increase in sFlt-1 levels outside of the eye. However, sFlt-1 levels were increased in the vitreous of the AAV.sFlt-1-injected eyes. Preliminary study of the immunogenicity of rAAV.sFlt-1 based on changes in the population of immune cell subsets by flow cytometry showed no significant changes in T-, B- and NK- cell and monocyte numbers.

Conclusions: : These preclinical studies have shown that subretinal administration of rAAV.sflt was well tolerated with no adverse effect on general health and visual function. There was a lack of any observed systemic toxicity or immunogenicity. In the light of these data and the long term efficacy of rAAV.sFlt-1 human trials are warranted.

Keywords: choroid: neovascularization • gene transfer/gene therapy • drug toxicity/drug effects 
×
×

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.

×