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Home / Equine Research Bank / Drug Test Anal / Discovery of Biomarkers of a Recombinant Human Erythropoieti...
Drug testing and analysis2024; doi: 10.1002/dta.3810

Discovery of Biomarkers of a Recombinant Human Erythropoietin Administration to Thoroughbred Geldings by Label-Free Proteomics.

Abstract: Erythropoiesis-stimulating agents (ESAs) continue to be a significant threat to the integrity of human and equine sports. Besides conventional direct testing, monitoring the biomarkers associated with the effects of ESAs may provide a complementary approach via indirect detection to enhance doping control. In this study, we applied label-free proteomics to discover plasma protein biomarkers in Thoroughbred geldings after administration with a long-acting form of recombinant human erythropoietin (rhEPO), methoxy polyethylene glycol epoetin beta, Mircera. Increased haematocrit, haemoglobin and red blood cell (RBC) levels were evidenced as early as 4 days post-administration in all three horses to varying extents. Tryptic peptides were obtained from plasma samples and analysed by nanoflow ultra-high-performance liquid chromatography-high-resolution tandem mass spectrometry (nano-UHPLC-HRMSMS) using data-independent acquisition. Differential protein abundance analysis has shortlisted seven protein biomarker candidates that showed significant changes specifically after Mircera administration in the treated but not in the control geldings, which comprised downregulation of two proteins, haptoglobin (HP) and haemopexin (HPX), and upregulation of five proteins, transferrin receptor 1 (TFR1), phospholipid transfer protein (PLTP), tenascin C (TNC), vascular cell adhesion molecule 1 (VCAM1) and galectin 3 binding protein (LGALS3BP). Multivariate analysis of plasma proteome has allowed the classification of control and treated samples. This is the first report on the discovery of plasma protein biomarkers of rhEPO administration to geldings. The results lay a foundation for applications of protein biomarkers for controlling the misuse of ESAs.
© 2024 John Wiley & Sons Ltd.
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Publication Date: 2024-09-22 PubMed ID: 39307543DOI: 10.1002/dta.3810Google Scholar: Lookup
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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.

The research identifies seven potential plasma protein biomarkers in racehorses to indirectly detect the use of erythropoiesis-stimulating agents (ESAs), substances that can be misused for performance enhancement in sports.

Context

  • ESAs increase the production of red blood cells, boosting oxygen delivery to muscles and improving athletes’ performance. However, they are prohibited in sports due to health risks and the potential for performance enhancement.
  • The traditional method of detecting ESAs misuse involves directly testing for the presence of the drugs. This research proposes a complementary approach by identifying biomarkers associated with the effects of ESAs through proteomic techniques.

Methodology

  • The researchers implemented a long-acting form of recombinant human erythropoietin (rhEPO), called Mircera, on Thoroughbred geldings (castrated male horses).
  • They observed the increase in haematocrit, haemoglobin, and red blood cell levels in all three horses as early as 4 days after administration, marking ESA’s effects.
  • Tryptic peptides were extracted from plasma samples and analysed using nanoflow ultra-high-performance liquid chromatography-high-resolution tandem mass spectrometry (nano-UHPLC-HRMSMS). This process allowed the researchers to identify and quantify the proteins present in the samples.

Findings

  • After the Mircera administration, seven biomarker candidates showed notable changes in the treated geldings but remained consistent in the control group.
  • Two proteins, haptoglobin (HP) and haemopexin (HPX), were reduced, while five others, including transferrin receptor 1 (TFR1), phospholipid transfer protein (PLTP), tenascin C (TNC), vascular cell adhesion molecule 1 (VCAM1), and galectin 3 binding protein (LGALS3BP), were boosted.
  • This differential protein abundance points to potential biomarkers of rhEPO administration and, more broadly, the misuse of ESAs.
  • Moreover, multivariate analysis of the plasma proteome successfully differentiated between samples from control and treated horses, which strengthens the findings.

Significance

  • This research offers new insights into plasma protein biomarkers associated with rhEPO administration in geldings. It represents a step towards a more comprehensive ESA misuse control approach in sports involving horses and potentially humans.
  • The identified biomarkers could help enhance existing testing methods, contributing to fair play and health protection in equine and human sports.

Cite This Article

APA
Cheung HW, Wong KS, Tam JCL, Farrington AF, Bond AJ, Wan TSM, Ho ENM. (2024). Discovery of Biomarkers of a Recombinant Human Erythropoietin Administration to Thoroughbred Geldings by Label-Free Proteomics. Drug Test Anal. https://doi.org/10.1002/dta.3810

Publication

Drug testing and analysis
ISSN: 1942-7611
NlmUniqueID: 101483449
Country: England
Language: English

Researcher Affiliations

Cheung, Hiu Wing
  • Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, Hong Kong, China.
Wong, Kin-Sing
  • Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, Hong Kong, China.
Tam, Jimmy C L
  • Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, Hong Kong, China.
Farrington, Adrian F
  • Veterinary Clinical Services, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, Hong Kong, China.
Bond, Amanda J
  • Equestrian Affairs, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, Hong Kong, China.
Wan, Terence S M
  • Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, Hong Kong, China.
Ho, Emmie N M
  • Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, Hong Kong, China.

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