ANALYTIC METHODS TO IDENTIFY MOLECULAR BIOMARKERS USEFUL TO DETECT GENE DOPING

During the last two decades, progress in Genetic, have been improperly applied by dishonest athletes, leading to a new frontier of doping: “Gene Doping”. This practice has been developed as collateral aspect of the gene therapy approach to diseases, and its dangerousness to health, is related to the fact that not certainty about the long-term effects of the use of gene manipulation, has been assessed. Another important problem is linked to the use of not controlled laboratories, leading to an increased risk of contamination by chemicals and other impurities during the production processes. For these reasons the anti-doping community has given extensive efforts on developing methods for gene doping detection which could be used by the World Anti-Doping Agency to monitor athletes. At the present, among the genes involved in physical activity and sports, those targeted in gene doping included ACE (angiotensin-converting enzyme), ACTN3 (actinin binding protein 3), EPO (erythropoetin), HGH (human growth factor), HIF (hypoxia inducible factor), IGF-I (insulin-like growth factor), PPAR-delta (peroxisome proliferators-activated receptor delta), VEGF (vascular endothelial growth factor), as well as Endorphins. Gene doping is very difficult to be detected. For example gene transfer, cannot be easily identified as the genomic material transfected is of the same origin of the parental one. Also the DNA sequence used for the transfection is difficult to be recognized as it is used only as a shuttle and therefore its level is very low. Even in the case of the analysis of the effect induced by gene doping, the analysis can be very invasive as, in some cases, a biopsy is required. To overcome these problems, new strategies were tentatively applied, which included proteomic analysis, based on both separation (liquid/gas chromatography) and mass spectrometry techniques.