Poster Presentation ASSCR, AGCTS, ISCT ANZ and Friends Joint Scientific Conference 2019

Human and mouse retinal models, and investigation of adeno-associated virus delivery (#128)

Steven S Eamegdool 1 , Arek Rybicki 2 , To Ha Loi 1 , Luisa M Osorio 1 , John Grigg 1 3 , Ulrike Grunert 3 , Michele C Madigan 3 , Leszek Lisowski 2 , Robyn V Jamieson 1 4
  1. Children's Medical Research Institute, Westmead, NSW, Australia
  2. Translational Vectorology Group, Children's Medical Research Institute, Westmead, NSW, Australia
  3. Save Sight Institute, University of Sydney, Sydney, NSW, Australia
  4. Discipline of Genomic Medicine, University of Sydney, Sydney, NSW, Australia

Introduction:

Heritable diseases of the retina are one of the primary causes of vision loss, with very few treatment options currently. Our gene-editing and replacement constructs require testing for efficiency and specificity of delivery for investigation of their therapeutic effects. We are assessing the delivery efficacy of adeno-associated viral (AAV) vector serotypes in our in vivo and in vitro model retinal systems.

 

Methods:

Wild-type ARC mice were subjected to both intravitreal and subretinal injections of AAV2 and AAV-7m8, with transduction monitored via fundoscopy, optical coherence tomography and immunofluorescence for 1 month. Retinal pigment epithelial (RPE) cells were also differentiated from human induced pluripotent stem cells (iPSCs) and transduced with AAV1-13. An interphase culture method was used to maintain human retinal explants ex vivo, which were exposed to AAV4 and AAV5.

 

Results and Discussion:

The mouse model system was used to assess whether the AAVs exhibited topographical transduction of the different retinal cell types. The iPSC-derived RPE cells were used to assess the transducibility of retinal cells in vitro, for application of this knowledge to retinal explants. Our combined approach allows selection of the most suitable AAVs with tropism to cells of interest for specific genetic retinal diseases.

  • Have you presented your abstract at another international meeting?: No