Submitter Withdrawn ASSCR, AGCTS, ISCT ANZ and Friends Joint Scientific Conference 2019

Targeting adeno-associated viral AAV vectors for systemic delivery to the skeleton (67888)

Lucinda R Lee 1 2 , Lauren Peacock 1 , Leszek Lisowski 3 4 , David G Little 1 2 , Craig F Munns 2 5 , Aaron Schindeler 1 2
  1. Orthopaedic Research and Biotechnology, The Children's Hospital at Westmead, Westmead, NSW, Australia
  2. Discipline of Child & Adolescent Health, Faculty of Health & Medicine, The University of Sydney, Sydney, NSW, Australia
  3. Translational Vectorology Group, Children's Medical Research Institute, Faculty of Medicine & Health, The University of Sydney, Sydney, NSW, Australia
  4. Military Institute of Hygiene and Epidemiology, The Biological Threats Identification and Countermeasure Centre, 24-100, Puławy, Poland
  5. Institute of Endocrinology & Diabetes, The Children's Hospital at Westmead, Westmead, NSW, Australia

Adeno associated viruses (AAVs) can selectively transduce different cell types although their potential for specific bone targeting remains underdeveloped. Screening AAV variants in a murine fracture model, we found that AAV8 and AAV-DJ can transduce bone cells at high levels. We generated cre recombinase expressing constructs under the control of CAG constitutive promoter or bone cell specific promoters Col2.3 or Sp7, packaged into AAV variants AAV2, AAV8, and AAV-DJ. These AAVs were systemically administered to Ai9 mice via tail vein delivery at a dose of 5×1010 and later 5×1011 vg/mouse. AAV8-CAG-cre was able to effectively lead to high levels of tdTomato reporter expression in osteoblasts following a single tail vein injection. Bone cell-specific promoters Col2.3 and Sp7 were able to effectively restrict cre recombinase expression to bone cells, with Sp7 proving to be more specific with very little expression in non-osseous tissues (including brain, heart, lungs, liver, spleen, and kidney). AAV8-Sp7-cre showed the highest efficiency and specificity for targeting bone cells of the skeleton following systemic delivery in mice. To conclude, this viral construct has scientific utility for creating bone-specific postnatal murine knockouts, and can be combined with CRISPR/Cas9 gene editing for functional genomics and gene therapy applications.

  • Have you presented your abstract at another international meeting?: Yes