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  • Sports Medicine for Anti-Doping
  • Next generation omics approaches in the fight against blood doping

Next generation omics approaches in the fight against blood doping

Despite being prohibited by World Anti-Doping Agency (WADA), blood manipulations such as the use of recombinant human erythropoietin (rHuEpo) and blood transfusions are well-known methods used by athletes to enhance performance. Direct detection of illicit blood manipulation has been partially successful due to the short detection window of the substances/methods, sample collection timing and the use of sophisticated masking strategies. In response, WADA introduced the Athlete Biological Passport (ABP) in 2009, which is an individualised longitudinal monitoring approach that tests primarily haematologic biomarkers of doping in order to identify atypical variability in response(s) in the athlete, highlighting a potential doping violation.

Although the implementation of the ABP has been an encouraging step forward in the quest for clean/drug-free sport, this detection method has some limitations. To reduce the risk of being detected by the ABP method, athletes are now resorting to microdoses of prohibited blood boosting substances to restrain abnormal fluctuations in the haematologic biomarkers, thereby reducing the sensitivity of the ABP detection method. Recent studies from numerous laboratories, including our own, have confirmed the potential of a transcriptomic microarray approach, which can assess distinct changes in gene expression after blood manipulations, to enhance the ABP. The closely interconnected studies conducted to date systematically evaluated an omics solution to detect blood doping with particular reference to rHuEpo doping. For example, relative to baseline, hundreds of genes were differentially expressed, that is they were switched ‘on’ and ‘off’, during rHuEpo administration. Notably, a few of these genes were switched ‘on’ after only a single rHuEpo injection, while further genes being switched ‘on’ during rHuEpo administration and remained switched ‘on’ up to four weeks after the last rHuEpo injection. A subset of these genes reflecting rHuEpo was further validated using a complementary gene expression technology and under different conditions such as blood transfusion, altitude training and exercise training to ensure the robustness of the technology and approach. These promising data provide the strongest evidence to date that omics technologies such as gene expression have the potential to add a new dimension to the ABP in terms of sensitivity and specificity for rHuEpo detection and blood doping in general.

Given the promising results, it is now of paramount importance to evaluate the effects of major confounding factors on this ‘molecular signature’ of rHuEpo doping, such as the effects of altitude and the effects of prior exercise. Specifically, there is an urgent need for developing specific and robust testing models which can differentiate altitude training from blood doping methods.

Therefore, the aim of this research is to compare blood gene expression profiles altered by rHuEpo (fairly high and microdose regimens, data from past and ongoing WADA-funded projects) and blood doping (with a focus on autologous transfusion) with altitude exposure in order to provide a set of candidate genes that can be used to detect blood doping.

Project timeframe

This research commenced in April 2017 and will end in March 2018.

Project aims

The aim of this research is to compare blood gene expression profiles altered by rHuEpo (fairly high and microdose regimens, data from past and ongoing WADA-funded projects) and blood doping (with a focus on autologous transfusion) with altitude exposure in order to provide a set of candidate genes that can be used to detect blood doping.

Project findings and impact

We believe that omics technologies integrated with more traditional methods offer the preferred method to strengthen the current ABP approach and contribute to other traditional anti-doping tests, especially for doping substances and methods difficult to detect, such as recombinant human growth hormone and blood transfusions.

In the main, anti-doping experts with hands-on experience of omics agree that the future of anti-doping research must reside in omics technologies. As for cancer research, most recent developments are firmly grounded in omics technologies despite numerous setbacks. In order to confirm that adding omics technologies to the ABP can significantly improve detection of rHuEPO and other difficult to detect drugs/methods, it is essential to developing specific and robust testing models which can differentiate altitude training from blood doping. Omics technologies such as gene expression have the potential to provide a ‘molecular signature’ specific to rHuEpo, blood transfusion and altitude exposure. The investigations and data acquisition proposed here are necessary before being able to assess the likelihood for inclusion of the omics biomarkers to the ABP and/or developing the next generation stand-alone test(s) to reveal doping or identify suspicious samples for targeting purposes. There is also the interesting possibility that this approach could help reduce the pressure on the athletes’ anti-doping obligations such as the ‘athletes' whereabouts’.

Research team

Professor Yannis Pitsiladis

Dr Guan Wang

Dr Alan Richardson

Antonia Karanikolou (PhD student)

Shaun Sutehall (PhD student)

Output

Karanikolou A, Wang G, Pitsiladis Y (2017) Letter to the editor: A genetic-based algorithm for personalized resistance training. Biol Sport 34:31-33.

Related output

Yannis Pitsiladis, Joanna Harper, Jonathan Ospina Betancurt, María-José Martínez Patiño, Attilio Parisi, Guan Wang, Fabio Pigozzi (2016) Beyond fairness: the biology of inclusion for transgender and intersex athletes. Current Sports Medicine Reports. 15(6): 386-388.

Wang G, Tanaka M, Eynon N, North KN, Williams AG, Collins M, Moran CN, Britton SL, Fuku N, Ashley EA, Klissouras V, Lucia A, Ahmetov II, de Geus E, Alsayrafi M, Pitsiladis YP. The Future of Genomic Research in Athletic Performance and Adaptations to Training. Med Sport Sci. 61:55-67.

Salamin O, Barras L, Robinson N, Baume N, Tissot JD, Pitsiladis Y, Saugy M, Leuenberger N (2016) Impact of blood transfusion on gene expression in human reticulocytes. Am J Hematol. Jul 7. doi: 10.1002/ajh.24470. [Epub ahead of print]

Jérôme Durussel, Diresibachew Haile, Kerli Mooses, Evangelia Daskalaki, Wendy Beattie, Martin Mooses, Wondyefraw Mekonen, Neford Ongaro, Edwin Anjila, Rajan Patel, Neal Padmanabhan, Martin McBride, John McClure, Yannis Pitsiladis (2016) The blood transcriptional signature of recombinant human erythropoietin administration and implications for anti-doping strategies. Physiological Genomics. Mar;48(3):202-9. doi: 10.1152/physiolgenomics.00108.2015. Epub 2016 Jan 12.

Pitsiladis YP, Tanaka M, Eynon N, Bouchard C, North KN, Williams AG, Collins M, Moran CN, Britton SL, Fuku N, Ashley EA, Klissouras V, Lucia A, Ahmetov II, de Geus E, Alsayrafi M; Athlome Project Consortium (2016) A Concerted Effort to Discover Genomic and other "OMIC" Markers of Athletic Performance. Physiological Genomics. Mar;48(3):183-90. doi: 10.1152/physiolgenomics.00105.2015. Epub 2015 Dec 29.

Pitsiladis YP, Durussel J, Rabin O (2014) An Integrative “Omics” Solution to the Detection of Recombinant Human Erythropoietin and Blood Doping. British Journal of Sports Medicine.May;48(10):856-61. doi: 10.1136/bjsports-2014-093529. Epub 2014 Mar 13.

Dvorak J, Saugy M, Pitsiladis YP (2014) Challenges and threats to implementing the fight against doping in sport. British Journal of Sports Medicine. May;48(10):807-9. doi: 10.1136/bjsports-2014-093589.

Dvorak J, Baume N, Botré F, Broséus J, Budgett R, Frey WO, Geyer H, Harcourt PR, Ho D, Howman D, Isola V, Lundby C, Marclay F, Peytavin A, Pipe A, Pitsiladis YP, Reichel C, Robinson N, Rodchenkov G, Saugy M, Sayegh S, Segura J, Thevis M, Vernec A, Viret M, Vouillamoz M, Zorzoli M (2014) Time for change: a roadmap to guide the implementation of the World Anti-Doping Code 2015. British Journal of Sports Medicine. May;48(10):801-6. doi: 10.1136/bjsports-2014-093561.

Jérôme Durussel, Evangelia Daskalaki, Martin Anderson, Tushar Chatterji, Diresibachew Haile, Neal Padmanabhan, Rajan K Patel, John D McClure, Yannis P Pitsiladis (2013) Effects of Recombinant Human Erythropoietin on Haemoglobin Mass, Blood Volume and Running Time Trial Performance in Trained Men. PLoS ONE. 2013;8(2):e56151. doi: 10.1371/journal.pone.0056151. Epub 2013 Feb 13.

Jérôme Durussel, Ramzy Ross, Prithvi R Kodi, Evangelia Daskalaki, Pantazis Takas, John Wilson, Bengt Kayser, Yannis P Pitsiladis (2013) Precision of the Optimized Carbon Monoxide Rebreathing Method to Determine Total Haemoglobin Mass and Blood Volume, European Journal of Sport Science, 13(1): 68-77, 2013.

Fabio Pigozzi, Alessia Di Gianfrancesco, Mario Zorzoli, Norbert Bachl, David Mc Donagh, Joseph Cummiskey, Luigi Di Luigi, Yannis Pitsiladis, Paolo Borrione (2012) Why glucocorticosteroids should remain in the list of prohibited substances: A sports medicine view point. International Journal of Immunopathology and Pharmacology. 25(1):19-24

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