| Autor: |
Brimble MA; Departments of, Host Microbe Interactions, St. Jude Children's Research Hospital, Memphis, Tennessee, USA., Morton CL; Departments of, Surgery, St. Jude Children's Research Hospital, Memphis, Tennessee, USA., Winston SM; Departments of, Surgery, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.; Graduate School of Biomedical Sciences, St. Jude Children's Research Hospital, Memphis, Tennessee, USA., Reeves IL; Departments of, Surgery, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.; Graduate School of Biomedical Sciences, St. Jude Children's Research Hospital, Memphis, Tennessee, USA., Spence Y; Departments of, Surgery, St. Jude Children's Research Hospital, Memphis, Tennessee, USA., Cheng PH; Departments of, Surgery, St. Jude Children's Research Hospital, Memphis, Tennessee, USA., Zhou J; Departments of, Surgery, St. Jude Children's Research Hospital, Memphis, Tennessee, USA., Nathwani AC; Research Department of Haematology, UCL Cancer Institute, University College London, London, United Kingdom., Thomas PG; Departments of, Host Microbe Interactions, St. Jude Children's Research Hospital, Memphis, Tennessee, USA., Souquette A; Department of Medicine, University of Maryland School of Medicine, Baltimore, Maryland, USA., Davidoff AM; Departments of, Surgery, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.; Graduate School of Biomedical Sciences, St. Jude Children's Research Hospital, Memphis, Tennessee, USA. |
| Abstrakt: |
Liver injury with concomitant loss of therapeutic transgene expression can be a clinical sequela of systemic administration of recombinant adeno-associated virus (rAAV) when used for gene therapy, and a significant barrier to treatment efficacy. Despite this, it has been difficult to replicate this phenotype in preclinical models, thereby limiting the field's ability to systematically investigate underlying biological mechanisms and develop interventions. Prior animal models have focused on capsid and transgene-related immunogenicity, but the impact of concurrently present nontransgene or vector antigens on therapeutic efficacy, such as those derived from contaminating nucleic acids within rAAV preps, has yet to be investigated. In this study, using Ad5-CMV_GFP-immunized immunocompetent BALB/cJ mice, and a coagulation factor VIII expressing rAAV preparation that contains green flourescent protein (GFP) cDNA packaged as P5-associated contaminants, we establish a model to induce transaminitis and observe concomitant therapeutic efficacy reduction after rAAV administration. We observed strong epitope-specific anti-GFP responses in splenic CD8+ T cells when GFP cDNA was delivered as a P5-associated contaminant of rAAV, which coincided and correlated with alanine and aspartate aminotransferase elevations. Furthermore, we report a significant reduction in detectable circulating FVIII protein, as compared with control mice. Lastly, we observed an elevation in the detection of AAV8 capsid-specific T cells when GFP was delivered either as a contaminant or transgene to Ad5-CMV_GFP-immunized mice. We present this model as a potential tool to study the underlying biology of post-AAV hepatotoxicity and demonstrate the potential for T cell responses against proteins produced from AAV encapsidated nontherapeutic nucleic acids, to interfere with efficacious gene transfer. |
