S52 Development of protocols for mouse GLP-toxicology studies

IntroductionWe have developed a Simian Immunodeficiency Virus (SIV)-based lentiviral vector pseudotyped with the Sendai-virus envelope glycoproteins (F/HN) (rSIV.F/HN) that is effective at transducing pulmonary epithelium in vivo. To prepare for a first-in-man clinical trial, we have developed protocols that can be used in a mouse GLP-toxicology study to efficiently transduce nasal-tissue or lungs.MethodsReporter-imaging, molecular, and radiopharmaceutical tracing methods have been used to quantify intranasally administered lentiviral vector deposition and subsequent tissue-level transgene expression in mice.ResultsNose study. A standard 100 µL vector bolus administration (‘nasal sniffing’) was compared to slow perfusion via catheter (6.7µL/min) and to small volume pipette dosing (2 × 5µL). All animals received 1e7 TU/mouse of rSIV.F/HN expressing luciferase (n=6/group). 10–12 days after transduction, all methods led to similar gene expression in the nasal cavity. Catheter-based delivery led to significant (p