Blood cells RNA biomarkers as a first long-term detection strategy for EPO abuse in horseracing.
Abstract: Recombinant human erythropoietins (rHuEPOs) are glycoproteins drugs, produced by the pharmaceutical industry to restore production of red blood cells by stimulating human bone marrow for which this pathology has been diagnosed. It is suspected that these molecules are diverted as doping agents in horseracing to enhance oxygen transport and aerobic power in racehorses. Although indirect double-blotting or direct liquid chromatography-mass spectrometry (LC-MS) methods have been developed to confirm the presence of rHuEPO in a sample, the short detection time (48 h) is still a problem for doping control. In this context, gene profiling investigation through Serial Analysis of Gene Expression (SAGE) has been conducted on seven thoroughbreds treated with Eprex. This functional genomic method has been performed from total blood cells collected from each animal to assess the mRNA expression consecutive to rHuEPO injections. Sample pooling was chosen as a powerful, cost-effective, and rapid means of identifying the most common and specific changes in terms of gene expression profile and to eliminate individual variation. Consequently, three SAGE libraries were constructed, before, during, and after Eprex treatment. More than 71 440 mRNA signatures were observed and subjected to statistical analysis; 49 differentially expressed genes were identified and analyzed by qPCR. From the selected gene list, were defined as potential biomarkers in terms of their low inter-individual variation and capacity as strong markers of rHuEPO administration up to 60 days after the beginning of the doping period. In this paper, a new strategy is proposed to the horseracing industry to prevent rHuEPO abuse.
Copyright 2010 John Wiley & Sons, Ltd.
Publication Date: 2010-08-06 PubMed ID: 20687115DOI: 10.1002/dta.146Google Scholar: Lookup
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- Journal Article
Summary
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The research article explores the development of a long-term detection strategy for the misuse of human erythropoietins (rHuEPOs) – medical substances used in doping – in horse racing.
Objective of the Research
- This research aimed to develop a long-term detection method for rHuEPO abuse in horseracing, which is an ongoing problem due to the short detection span (48 hours) of current methods. rHuEPOs stimulate red blood cells’ production, and they are often misused to improve endurance in racehorses.
The Detection Process Used
- The researchers conducted gene profiling studies using Serial Analysis of Gene Expression (SAGE) on seven thoroughbred horses treated with a form of rHuEPO called Eprex.
- They collected total blood cells from each animal to assess the changes in mRNA expression due to rHuEPO injections.
- Instead of individual sample analysis, they pooled the samples to establish a common gene expression profile changes which is also an economical and speedy method.
Construction of SAGE Libraries
- The researchers structured three SAGE libraries representing different stages: before, during, and after Eprex treatment.
- They observed more than 71,440 mRNA signatures in these libraries and conducted a statistical analysis of these results.
Identification of Potential Biomarkers
- From the analysis, they identified 49 differentially expressed genes which they further investigated using qPCR.
- Subsequently, certain genes were chosen as potential biomarkers based on their low inter-individual variation and their effectiveness as rHuEPO administration indicators for up to 60 days from the initial doping period.
Proposal of New Strategy
- The research suggests this new strategy to the horseracing industry for preventing rHuEPO abuse, by allowing for a longer detection window, which improves the chances of identifying any form of doping.
Cite This Article
APA
Bailly-Chouriberry L, Noguier F, Manchon L, Piquemal D, Garcia P, Popot MA, Bonnaire Y.
(2010).
Blood cells RNA biomarkers as a first long-term detection strategy for EPO abuse in horseracing.
Drug Test Anal, 2(7), 339-345.
https://doi.org/10.1002/dta.146 Publication
Researcher Affiliations
- LCH, Laboratoire des Courses Hippiques, 15 rue de Paradis, 91370 Verrières-le-Buisson, France. l.bailly@lchfrance.fr
MeSH Terms
- Animals
- Biomarkers / blood
- Doping in Sports
- Epoetin Alfa
- Erythropoietin / administration & dosage
- Erythropoietin / analysis
- Female
- Gene Expression Profiling / methods
- Horses
- Humans
- Male
- Polymerase Chain Reaction / methods
- RNA, Messenger / blood
- Recombinant Proteins
- Substance Abuse Detection / methods
Citations
This article has been cited 3 times.- Dahlgren AR, Knych HK, Arthur RM, Durbin-Johnson BP, Finno CJ. Transcriptomic Markers of Recombinant Human Erythropoietin Micro-Dosing in Thoroughbred Horses. Genes (Basel) 2021 Nov 24;12(12).
- Spornraft M, Kirchner B, Haase B, Benes V, Pfaffl MW, Riedmaier I. Optimization of extraction of circulating RNAs from plasma--enabling small RNA sequencing. PLoS One 2014;9(9):e107259.
- Loria F, Grabherr S, Kuuranne T, Leuenberger N. Use of RNA biomarkers in the antidoping field. Bioanalysis 2024;16(10):475-484.
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