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Drug testing and analysis2024; 17(7); 1078-1087; doi: 10.1002/dta.3812

New Transcriptomic Biomarkers for Detection of the Recombinant Human Erythropoietin (rHuEPO) MirCERA in Horses.

Abstract: Detection and monitoring of biomarkers related to doping is particularly suitable for the development of analytical strategies dedicated to indirect detection of banned substances. Previous studies in horses have already allowed the investigation of transcriptomic biomarkers in equine blood associated with reGH and rHuEPO administrations. Our most recent developments continue to focus on the discovery and monitoring of transcriptomic biomarkers for the control of ESAs, and a collaborative study with WADA-accredited doping control laboratories has recently been initiated to conduct a pilot study. In humans, three mRNAs (ALAS2, CA1, and SLC4A1) were previously observed to be differentially expressed after blood doping and were associated with immature red blood cells, the so-called circulating reticulocytes. In horses, circulating reticulocytes are rarely observed even after rHuEPO administration. With the improved primers that detect the equine orthologues of the human mRNAs from the ALAS2, CA1, and SLC4A1 genes, we can now report the first evidence of the detection of the three biomarkers in equine blood. In addition, an upregulation of the mRNA levels of the three genes was observed after analysis of blood samples collected from MirCERA-treated animals, with kinetics similar to those previously documented in humans. Our data suggest that ALAS2 and CA1 are promising indirect biomarkers for the detection of recombinant EPO abuse in horses, as observed in humans.
© 2024 John Wiley & Sons Ltd.
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Publication Date: 2024-09-25 PubMed ID: 39321850DOI: 10.1002/dta.3812Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigates the detection and monitoring of doping in horses using biomarkers, particularly focusing on the drug Recombinant Human Erythropoietin (rHuEPO) MirCERA. Results showed that there was an increase in the mRNA levels of three genes (ALAS2, CA1, and SLC4A1) in the blood of MirCERA-treated horses, similar to previous findings in humans.

Background and Objectives

  • The main objective of this research was to develop better methodologies for detecting the use of banned substances in horses, focusing on rHuEPO MirCERA, and to monitor these using transcriptomic biomarkers.
  • The study aimed to further build on the work of previous studies which had explored using transcriptomic biomarkers found in equine blood associated with the administration of rHuEPO.
  • The research also sought to provide an improved primer for detecting these biomarkers.

Methodology

  • The study utilized the three mRNAs (ALAS2, CA1, and SLC4A1), which were previously observed to be differentially expressed after blood doping, associated with immature red blood cells or circulating reticulocytes.
  • Using these mRNAs as potential biomarkers, the study developed improved primers to detect the existence of these biomarkers in equine blood.
  • The analysis of blood samples from MirCERA-treated horses was done to see if there was a change in the mRNA levels of the three genes.

Findings

  • After analysis of the blood samples, it was discovered that an upregulation of the mRNA levels of the three genes was observed in MirCERA-treated animals.
  • Interestingly, the kinetics for this occurrence were similar those documented in previous studies involving humans.
  • The study concluded that, specifically, ALAS2 and CA1 genes were promising as indirect biomarkers for the detection of rHuEPO abuse in horses.

Significance and Implications

  • This study offers the first-ever evidence that these three specific biomarkers can be detected in equine blood.
  • It not only advances our understanding of drug detection in sports animals but may also lead to more robust and accurate anti-doping measures in the future.
  • The research could play a crucial role in collaborative projects with authoritative bodies such as the World Anti-Doping Agency (WADA), enhancing monitoring and control of potential doping in sports involving animals.

Cite This Article

APA
Loup B, André F, Leuenberger N, Marchand A, Barnabé A, Delcourt V, Garcia P, Popot MA, Bailly-Chouriberry L. (2024). New Transcriptomic Biomarkers for Detection of the Recombinant Human Erythropoietin (rHuEPO) MirCERA in Horses. Drug Test Anal, 17(7), 1078-1087. https://doi.org/10.1002/dta.3812

Publication

Drug testing and analysis
ISSN: 1942-7611
NlmUniqueID: 101483449
Country: England
Language: English
Volume: 17
Issue: 7
Pages: 1078-1087

Researcher Affiliations

Loup, Benoit
  • GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
André, François
  • GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
Leuenberger, Nicolas
  • Swiss Laboratory for Doping Analyses, University Center of Legal Medicine, Lausanne and Geneva, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
Marchand, Alexandre
  • Laboratoire antidopage français (LADF), Université Paris-Saclay, Orsay, France.
Barnabé, Agnès
  • GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
Delcourt, Vivian
  • GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
Garcia, Patrice
  • GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
Popot, Marie-Agnès
  • GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
Bailly-Chouriberry, Ludovic
  • GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.

MeSH Terms

  • Horses / blood
  • Horses / genetics
  • Animals
  • Doping in Sports
  • Erythropoietin / blood
  • Erythropoietin / administration & dosage
  • Biomarkers / blood
  • Substance Abuse Detection / methods
  • Substance Abuse Detection / veterinary
  • Recombinant Proteins / blood
  • Humans
  • RNA, Messenger / blood
  • RNA, Messenger / genetics
  • Transcriptome
  • 5-Aminolevulinate Synthetase / genetics
  • 5-Aminolevulinate Synthetase / blood
  • Anion Exchange Protein 1, Erythrocyte / genetics
  • Anion Exchange Protein 1, Erythrocyte / blood

Grant Funding

  • Institut Français du Cheval et de l'Equitation

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