Projets de recherche

The proposed project will identify and characterize sportspecific stressors contributing to doping vulnerability among athletes, to include exploring their impact, frequency, severity, timing, and duration. Additionally, it seeks to investigate how psychological flexibility moderates the relationship between these stressors and doping vulnerability, thereby protecting athletes from doping.

A qualitative approach and informed by a co-design process with national anti-doping and sporting organizations. This project seeks to interview women in three countries to 1) investigate the role of support personnel (e.g., coaches, trainers, partners) in facilitating or preventing PIED use, and 2) to understand the broader psycho-socio-cultural factors that contribute to facilitating or preventing PIED use among female athletes. Investigating when, why, and how women choose to use or avoid PIEDs this project will gain important insights on women’s lived experiences with PIEDs.

A cross-sectional survey design and recruit a large sample of elite athletes (1500) from the said countries. The survey will include a number of validated screening tools in Spanish about self-compassion, mindfulness, sport anxiety, perfectionism, and doping susceptibility. Participants will also be invited to take part in a podcast series and to share their insights and experiences on doping in sport. In order to contribute towards more informed decision-making playing a crucial role in the prevention of doping behavior in the PANRADO Region, while focusing on a positive psychology approach emphasizing the role of mindfulness and selfcompassion.

Code: 23A20MP

GC-C-IRMS is used to confirm the exogenous origin of endogenous anabolic steroids. Samples are analyzed with a fast initial testing procedure to isolate suspicious samples and, afterwards, the suspicious samples are analyzed with an IRMS confirmatory method. High throughput IRMS will allow a substantial expansion of CIR analyses, enabling a considerable increase in doping violation detections and the set-up of an efficient CIR ABP module. High throughput IRMS is also highly desirable in those cases where a large number of samples needs to be tested on IRMS in relative short period of time e.g., preseason/regular season testing and Olympic Games.

Code: 23B02CR

Chapter S2 of WADA’s Prohibited List 2023 (“Peptide hormones, growth factors, related substances, and mimetics”) lists Erythropoietin Receptor Agonists (ERAs) and Transforming growth factor beta (TGF-ß) signalling inhibitors (luspatercept, sotatercept) under chapter 1 (“Erythropoietins (EPO) and agents affecting erythropoiesis”) as prohibited substances. Currently, SAR- and SDS-PAGE are the most frequently applied techniques for the initial testing and confirmation procedures (ITP, CP) for ERAs in WADA accredited laboratories worldwide (TD2022EPO). While electrophoretic detection methods for TGF-ß signalling inhibitors were also developed, they are not yet part of the technical document. In 2019 and 2020, the first luspatercept-based pharmaceutical (Reblozyl®, “luspatercept-aamt”) was approved by FDA (USA) and EMA (Europe).

Hence, routine testing for TGF-ß signalling inhibitors will become necessary in the near future. For the detection of erythropoietins and TGF-ß signalling inhibitors in blood and urine immunoaffinity purification is required before electrophoretic separation. Due to significant structural differences between these compounds, three different capture antibodies have to be employed, i.e. anti-EPO, anti-activin receptor type IIA (ACVR2A), and type IIB (ACVR2B) antibodies for epoetins, sotatercept and luspatercept, respectively. A protocol for the combined immunopurification of ERAs, sotatercept and luspatercept followed by isoelectric focusing (IEF-PAGE) was published in 2019, another one for ERAs and luspatercept in combination with SDS- and SAR-PAGE in 2021. Both protocols were based on covalent immobilization of relatively large amounts of the capture antibodies on magnetic beads.

Consequently, the beads had to be re-used several times for cost-reduction. Additionally, sotatercept was not included in the protocol for SDS- and SAR-PAGE. We already developed individual protocols for capturing sotatercept and luspatercept in serum samples, which use non-covalent immobilization of very small antibody amounts on anti-antibody coated magnetic beads. Additionally, we presented a similar cost-minimized protocol for the purification of EPOs from blood and urine at the Cologne workshop in March 2023. The plan is to combine these protocols in order to simultaneously purify all three compounds from the three sample matrices (serum/plasma/urine). Another disadvantage of the 2021 protocol was, that EPOs and luspatercept could not be detected in a truly multiplexed way after electrophoretic separation and Western blotting.

The reason were interferences caused by non-specific binding of the two detection antibodies with co-eluted proteins (the protocol used the same polyclonal antibody for capture and detection of luspatercept). Hence, the membrane had to be first incubated with an anti-EPO antibody followed by re-incubation with the anti-ACVR2B antibody. Contrary to that, the proposal of this project is that (1) the target proteins will be simultaneously detected by incubating the blot membrane with a mixture of all three detection antibodies, (2) it will also include sotatercept, and (3) the protocol will also be applicable to urine (it was shown in 2022 that luspatercept is also partly excreted in urine).

Code: 23E04FP

It is well known that the use performance-enhancing substances by athletes provide an unfair advantage and doping is contrary to the ‘the spirit of sport’. However, a randomised response surveys conducted between 2011-2018 indicate that >15% of athletes have doped with banned substances. Yet in 2018, WADA reported only 2% positive samples, with hormone doping accounting for >50% of adverse analytical findings. These statistics suggests that micro-dosing, off-competition doping and cyclical (on/off)/pyramiding patterns of administering anabolic steroids (AAS) have allowed athletes to evade current detection methods, while beneficial for building muscle mass, strength and performance. In addition, drug administration studies in humans have demonstrated that testosterone dose-dependently increases satellite cell and myonuclei number, muscle fibre cross sectional area (CSA) and induces hypertrophy of muscle fibres. These benefits may remain long after cessation of AAS use and/or detraining leading a muscle atrophy, when a new stimulus (i.e., returning to training) is given, the performance is enhanced, a phenomenon known as “muscle memory”. The “muscle memory” associated with AAS could explain why androgen cycling has been so effective for doping athletes. AAS treatment in mice for 14 days has been shown to increase myonuclei number by 66%, and these are retained despite drug withdrawal and contribute to faster muscle growth in response to muscle overload (Egner et al, 2013). These findings indicate that even brief exposure to AAS may provide long-term benefits for muscle performance. Increasing understanding of the mechanisms associated with AAS muscle memory is thus vital for enhancing anti-doping detection. Other mechanisms, besides myonuclei accretion/retention, have also been implicated with the muscle memory phenomenon, and the understanding of this process is crucial for the identification of long-term biomarkers. Therefore, the aim of this research is to identify relevant biomarkers associated with past-AAS use. Building on previous work, we will generate and characterise a mouse model of AAS muscle memory and explore the associated mechanisms using transcriptomic, phosphoproteomic and metabolomic approaches. In our previous research funded by a WADA research grant (16E11FP), we have established a tissue biobank (muscle biopsies, saliva, urine, serum) from powerlifters that either do not use AAS, actively use AAS or have previously used AAS. Guided by findings in mice, we will examine the utility of these markers for identifying current and past-AAS use in our biobank of tissues. Our findings are envisaged to enhance anti-doping detection, provide guidance for appropriate punitive measures in response to positive tests, and contribute to ongoing debates regarding transgender athlete inclusion in elite competition.

Code: 23D10YD

Our main purpose of this study is to evaluate non-analytical factors that may influence the markers in the serum steroid profiles and endocrine profiles, which are indirect methods to detect doping with anabolic androgenic steroids and growth hormone, respectively, and which will be integrated into the WADA Athlete Biological Passport (ABP) in 2023. The new knowledge will support the interpretation and evaluation of serum steroid and endocrine profiles, and may increase the sensitivity of the ABP. In project 1, 27 Swedish athletes with symptoms of long-term low energy availability (LEA) and 27 matched controls are invited to participate in a cross-sectional study evaluating the influence of long-term LEA on the serum endocrine and steroid profiles in athletes. It is hypothesized that male and female athletes with long-term LEA expressed as reproductive dysfunction (amenorrhea/low libido) will have suppressed levels of present and new biomarkers in the serum steroid and endocrine profiles compared with male and female athletes with normal reproductive function without symptoms of eating disorders. In project 2, 22 female national level powerlifters and weightlifters in Norway are recruited to evaluate changes in endocrine and steroid profiles in serum after a familiar resistance training session at two different time points (follicular phase and luteal phase) in the menstrual cycle. Based on existing literature, it is hypothesized that present and new serum steroid biomarkers are unaltered and that endocrine biomarkers are increased and more variable immediately after a resistance training session in female weightlifters and powerlifters during either menstrual cycle phases.

Code: 23C07KD

Tesamorelin is a synthetic growth hormone-releasing hormone (GHRH) analogue, used for the reduction of excess abdominal fat in HIV-infected adults. As a GHRH analogue, it triggers the secretion of growth hormone (GH), which in turn increases serum insulin-like growth factor 1 (IGF-1) levels. The latter exerts an anabolic (growth) effect on tissues throughout the human body. Since this anabolic effect is evidently beneficial for many athletes, the GHRH – GH –IGF-1 axis is a common target in sports doping. All three compounds, as well as their analogues, are banned by the World Anti-Doping Agency (WADA) at all times. Besides curating the list of prohibited compounds, WADA also provides doping control laboratories with a set of minimum required performance levels (MRPL) for these compounds. These MRPLs indicate the sensitivity that a detection method should achieve for a certain compound and are commonly based on administration studies. Since doping often corresponds to the illicit use of prescription medicine, administration studies for many prohibited compounds have only been performed in the context of a clinical trial involving a specific patient population. This research project aims to perform a Tesamorelin administration study on six healthy volunteers. This will allow for a re-evaluation of the current MRPL using a study population more representative of healthy athletes, as well as generate insight into the urinary excretion pattern of Tesamorelin. Both are important factors for the anti-doping community in evaluating the performance of detection methods. For the second objective of this project, the influence of Tesamorelin administration on serum IGF-1 levels will be monitored to investigate the possibility of using IGF-1 as a long-term marker for GHRH doping. Also P-III-NP levels will be assessed to evaluate if the IGF-1/P-III-NP ratio (as will be applied in the endocrine module of the athlete biological passport) is a stronger marker than IGF-1 alone to detect Tesamorelin abuse and GH secretagogues in general in sport.

Code: T23A05GG

The project deal with the synthesis and analytical characterization of an epimeric long-term metabolite of the anabolic steroid Dehydrochloromethyltestosterone (4a-chloro-18-nor-17b-hydroxymethyl-17a-methyl-5a-androst-13-en-3a-ol)

Code: T23A03GG

The project deal with the synthesis and analytical characterization of a long term metabolite of the anabolic steroid oxymesterone (17β-methyl-4,17α-dihydroxy-androst-4-en-3-one)