mRNA biomarkers sensitive and specific to micro-dose erythropoietin treatment at sea level and altitude

Francesco Loria*, Andreas Breenfeldt Andersen, Jacob Bejder, Thomas Bonne, Silke Grabherr, Tiia Kuuranne, Nicolas Leuenberger*, Nikolai Baastrup Nordsborg*

*Corresponding author af dette arbejde

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

1 Citationer (Scopus)

Abstract

Recombinant human erythropoietin (rhEPO) is prohibited by the World Anti-Doping Agency. rhEPO abuse can be indirectly detected via the athlete biological passport (ABP). However, altitude exposure challenges interpretation of the ABP. This study investigated whether 5′-aminolevulinate synthase 2 (ALAS2) and carbonic anhydrase 1 (CA1) in capillary dried blood spots (DBSs) are sensitive and specific markers of rhEPO treatment at altitude. ALAS2 and CA1 expression was monitored in DBS collected weekly before, during, and after a 3-week period at sea level or altitude. Participants were randomly assigned to receive 20 IU kg bw−1 epoetin alpha (rhEPO) or placebo injections every second day for 3 weeks while staying at sea level (rhEPO, n = 25; placebo, n = 9) or altitude (rhEPO, n = 12; placebo, n = 27). ALAS2 and CA1 expression increased up to 300% and 200%, respectively, upon rhEPO treatment at sea-level and altitude (P-values <0.05). When a blinded investigator interpreted the results, ALAS2 and CA1 expression had a sensitivity of 92%. Altitude did not confound the interpretation. Altitude affected ALAS2 and CA1 expression less than actual ABP markers when compared between sea level and altitude results. An individual athlete passport-like approach simulation confirmed the biomarker potential of ALAS2 and CA1. ALAS2 and CA1 were sensitive and specific biomarkers of micro-dose rhEPO treatment at sea level and altitude. Altitude seemed less a confounding factor for these biomarkers, especially when they are combined. Thus, micro-dose rhEPO injections can be detected in a longitudinal blinded setting using mRNA biomarkers in DBS.

OriginalsprogEngelsk
TidsskriftDrug Testing and Analysis
ISSN1942-7603
DOI
StatusAccepteret/In press - 2024

Bibliografisk note

Funding Information:
This project has been carried out with the support of the Dried Blood Spot Project. The Dried Blood Spot Project is a collaborative project between the World Anti‐Doping Agency (WADA) (Grant No. R17R02NN), the International Olympic Committee (IOC), the China Anti‐Doping Agency (CHINADA), the International Testing Agency (ITA) and the U.S. Anti‐Doping Agency (USADA), with additional financial support by the Anti‐doping Switzerland (ADCH), Sport Integrity Australia (SIA), and the Japan Anti‐Doping Agency (JADA).

Publisher Copyright:
© 2024 John Wiley & Sons Ltd.

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