Use of normo-thermic perfused organs to profile PK and infection efficiency of novel viral gene therapy vectors.

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Major barriers to effective gene therapy include rapid neutralisation in the blood stream, high Kupffer cell capture in the liver and low efficiency of target cell infection. Targeted genetic modification can generate lentiviral vector variants which may have the potential to overcome these barriers. However, without valid test systems with which to compare new variants, misleading data may result in unsuitable variants being prioritised for clinical development. Indeed, current pre-clinical models fail to adequately recapitulate the scale and physiology of human patients. Rodents are commonly used but are a particularly poor model, with a circulation rate 13x faster than humans, liver endothelial fenestrae gaps 30-60nm wider than in humans [Wisse,2008,GT], a blood volume and total mass 3000x lower than humans, and a very different history of virus pre-exposure and blood cell complement receptor profile [Carlisle,2009,Blood]. We are uniquely positioned to be able to provide a system, a normothermic perfusion device (NTPD), which can maintain human organs outside the body and permit their dosing with therapeutic agents. Continuous perfusion allows for blood and bile samples to be taken so that organ function/toxicity and viral vector pharmacokinetics can be defined, whilst sampling for virus genomes and transgene products by punch tissue biopsies taken at regular junctures can also track rates and levels of entry and integration. We propose that, in collaboration with Oxford Biomedica, we will use the NTPD to profile the PK of novel Lentiviral vector variants developed in Oxford Biomedica.