Loup Benoit, André François, Leuenberger Nicolas, Marchand Alexandre, Barnabé Agnès, Delcourt Vivian, Garcia Patrice, Popot Marie-Agnès, Bailly-Chouriberry Ludovic
GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
Swiss Laboratory for Doping Analyses, University Center of Legal Medicine, Lausanne and Geneva, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
Drug Test Anal. 2024 Sep 25. doi: 10.1002/dta.3812.
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.
与兴奋剂相关的生物标志物的检测和监测特别适合用于开发专门用于间接检测违禁物质的分析策略。先前对马的研究已经能够调查与重组生长激素（reGH）和重组人促红细胞生成素（rHuEPO）给药相关的马血液中的转录组生物标志物。我们最近的进展继续聚焦于发现和监测用于促红细胞生成素类似物（ESA）控制的转录组生物标志物，并且最近已经启动了一项与世界反兴奋剂机构（WADA）认可的兴奋剂检测实验室的合作研究来开展一项试点研究。在人类中，先前观察到三种信使核糖核酸（mRNA，即ALAS2、CA1和SLC4A1）在血液兴奋剂使用后表达存在差异，并且与未成熟红细胞，即所谓的循环网织红细胞有关。在马中，即使给予rHuEPO后也很少观察到循环网织红细胞。利用改进后的引物，能够检测来自人类ALAS2、CA1和SLC4A1基因的马直系同源基因，我们现在可以报告在马血液中检测到这三种生物标志物的首个证据。此外，在分析从经米卡芬净（MirCERA）处理的动物采集的血液样本后，观察到这三个基因的mRNA水平上调，其动力学与先前在人类中记录的相似。我们的数据表明，正如在人类中观察到的那样，ALAS2和CA1有望成为检测马中重组促红细胞生成素滥用的间接生物标志物。
