Projets de recherche

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Code: 21E04AB

This year WADA published the first version of Guidelines for Gene Doping Detection. The Guidelines apply to testing for doping genes using polymerase chain reaction (PCR). A PCR-based test developed in our laboratory and approved by WADA to be included in doping control has high sensitivity and is easy to use. Yet, as technologies for genetic measurements continue to develop, anti-doping scientists are employing them to develop alternative methods for gene doping testing.

Recently, we developed a novel method for gene doping detection based on massively parallel sequencing (MPS). This technology is rapidly expanding into clinical care with potential applications in diagnostics and therapeutics. The conceivable strength of the MPS methodology in gene doping testing is that it provides sequence-based information, which may be more difficult to challenge legally.

The current project aims to further validate the developed MPS method that simultaneously targets five genes, likely candidates for doping, but can be adapted for other doping genetic material. We will validate the full experimental protocol, the standard material that was designed and produced for quality controls, and the sophisticated bioinformatics pipeline that was developed for the test. We will also optimise DNA extraction from blood to improve recovery and purity of doping genes to increase reliability of the test. By analysing identical samples using the MPS- and PCR-based methods, we will compare the two tests for detecting five doping genes in terms of sensitivity, cost, ease and reliability.

We expect that on completion of the project and using our experience in developing a gene doping test capable to meet WADA’s accreditation requirements, a prototype test for gene doping detection based on MPS technology will be developed. Project’s outcomes will assist WADA in formulating a plan to transfer the method to WADA-accredited laboratories for implementation in routine anti-doping testing.

Code: 21C14AR

This project i a three-armed randomised controlled trial investigating the effect of WADA approved anti asthmatic medication on sprint performance during submaximal cycling. In addition to performance testing we will analyse urine and dried blood spot samples produce reference values for approved use of the respective medication. We will administer the newly WADA approved beta2 agonist vilanterol in combination with flutikasonfuroat, beclomethasone or placebo to 45 non asthmatic well-trained cyclists or triathletes. Performance test and analysis of urine will be performed after a single dose and one week’s use.

The study will address knowledge gaps regarding the effect approved doses of beta2 agonists identified by meta-analysis published in 2020 and knowledge gaps identified by a systematic review of regarding glucocorticoid administration in athletes. Our research will contribute to establishing thresholds to distinguish approved from prohibited use and develop new methods for analysis. *Research hypothesis and objectives The overall aims of the present study are to investigate the effect the of WADA approved anti-asthmatic medication vilanterol and beclomethasone on physical performance in health athletes, and to investigate the concentration levels and inter-subject variability in both urine and dried blood spots for establishment of levels consistent with therapeutic use.

In detail vilanterol and beclomethasone will be investigated to asses:

1. the effect on anaerobic performance during and after prolonged sub-maximal endurance exercise in well trained non-asthmatic athletes

2. the urine concentrations after prolonged endurance activity in athletes after acute and short-term daily inhalation of therapeutic doses

3. the utility of dried blood spots for detection of approved use after prolonged endurance activity

Code: 21C08XT

WADA drew the attention of the Laboratories to the possible detection of some anabolic androgenic steroids (AAS) and/or hormone and aromatase inhibitors in urine samples resulting from

the in situ microbial transformations of endogenous steroids in its Technical Letter TL20. The document establishes some requirements to verify if the findings could originate from microbial activity when the involved compounds are detected at concentrations below the MRPL. However there not enough scientific evidences or literature data to support this. The overall objective of the current project is to produce evidences to better support the actions to be taken when detecting 1-ene steroids metabolites, 7-oxo-steroids or 2- & 3-ene-steroids in human urine at low concentrations and to disclose the origin of these metabolites (external administration or produced by microbial contamination) as expected following WADA TL20. To reach the objectives the project intends to confirm specific metabolites for the 1-ene-5a-reduced steroids not originated from microbial contamination, produce specific metabolites of arimistane and 7-keto-DHEA, study the specificity of Δ1-steroid dehydrogenase over different substrates and finally study the formation of delta-2 (5a-androstane-2-en-17-one) metabolites excretion and their stability in human urine. This should allow a better evaluation of data produced by the WADA accredited Laboratories.

Code: 21A13GJ

Beta2-agonists salbutamol, formoterol, terbutaline and salmeterol delivered by inhalation are among the most used medications in athletes for treating asthma and exercise induced bronchoconstriction. To prevent supratherapeutic misuse that could result in performance enhancing effects, maximum permitted doses have been introduced for salbutamol and formoterol which are enforced through a urine threshold that corresponds to the maximum permitted dose. However, urine concentrations can be affected by a range of factors such as hydration status and urine production leading to large variations in urine concentrations of drug, both inter-patient, and intra-patient, which makes refining the urine threshold difficult. An alternative approach for quantifying beta2-agonist doping is measurement of concentrations using dried blood spots and dried plasma spots. These are easily conducted via finger prick, require no specialized personnel, are minimally invasive, and easily shipped to laboratories due to their light weight. The present project aims to investigate the utility of dried blood spots and dried plasma spot approaches for the measurement of systemic concentrations of the commonly used beta2-agonist salbutamol, formoterol, terbutaline, and salmeterol. Subjects will inhale beta2-agonists while exercising over a 1.5 h period with repeated sampling. Blood concentrations of beta2-agonists will be measured by dried blood spots and validated against traditional venous blood and urine concentrations. It is hoped that dried blood spots and dried plasma spots will avoid the variability traditionally associated with urine concentrations and help refine the detection of supratherapeutic dosing of beta2-agonists.

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Doping occurs in a complex and dynamic environment. Often, the sporting system in which doping occurs comprises multiple hierarchical levels that is challenging to fully elucidate. To date, research in doping has typically focused on athletes and athlete support personnel at the sharp end of this hierarchy rather than the broader systemic factors that contribute to doping. To understand and optimise the behaviour of complex systems (i.e., the anti-doping system), the application of contemporary systems thinking-based methods has been recommended. Such methods allow for analytical modelling to be conducted to understand and identify the numerous interacting factors that influence system behaviour. This enables the identification of opportunities to intervene and optimise the overall system rather than its component parts.