Unequivocal detection of AAV-mediated gene doping by pan-AAV immunological profiling

Code: ISF17E07FB 

The rationale of our proposal is based on the hypothesis that gene transfer with AAVs will leave an immunological and genetic footprint in any human subject clearly distinguishable from a naturally occurring AAV infection with the wild-type virus. In the context of the detection of gene doping, the main objective of this project is to develop a two-step method for the detection of AAV-mediated gene doping based on the combination of immunological and molecular tools. This general objective can be divided in the following specific aims, describing the two-step approach proposed for the identification of subjects who underwent AAV-based gene doping. Specifically: 2.1. Profiling of the anti-AAV antibody responses. A sensitive, pan-AAV antibody assay will be developed, validated, and streamlined to distinguish humoral responses deriving from natural exposure to AAV from gene doping with AAV vectors. Thus, this assay will permit to establish a baseline to be used as reference value to discriminate between subjects positive or negative for AAV-gene doping. 2.2. Detection of AAV genomes in blood/PBMCs by means of target capture NGS. To this end, a panel will be designed containing at least 4 million capture probes specific for at least 100 genome elements, such as viral ITs, regulatory sequences and different doping genes involved in improvement of muscle strength and performance.