Serum Proteomic Changes after Randomized Prolonged Erythropoietin Treatment and/or Endurance Training: Detection of Novel Biomarkers

• Published in: PloS one Vol. 10; no. 2; p. e0117119
• Main Authors: Christensen, Britt, Ludvigsen, Maja, Nellemann, Birgitte, Kopchick, John J., Honoré, Bent, Jørgensen, Jens Otto L.
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 13.02.2015; Public Library of Science (PLoS)
• Subjects: Abuse; Adolescent; Adult; Biological markers; Biomarkers; Biomarkers - blood; Blood; Blood Proteins; Chromatography, Liquid; Darbepoetin alfa - therapeutic use; Electrophoresis; Electrophoresis, Gel, Two-Dimensional; Endocrinology; Endurance; Erythrocyte Indices; Erythropoiesis; Erythropoietin; Erythropoietin - therapeutic use; Exercise; Gel electrophoresis; Gene expression; Haptoglobin; Haptoglobins; Hematology; Hospitals; Humans; Internal medicine; Laboratories; Male; Males; Mass spectrometry; Mass spectroscopy; Medical research; Medicine; Metabolism; Molecular chains; Musculoskeletal system; Oxygen Consumption; Passports; Physical Endurance; Physical training; Proteins; Proteome; Proteomics; Serum proteins; Spots; Studies; Tandem Mass Spectrometry; Training; Urine; Washout; Young Adult; Denmark; Ohio; United States > US; Aarhus Denmark
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0117119

Despite implementation of the biological passport to detect erythropoietin abuse, a need for additional biomarkers remains. We used a proteomic approach to identify novel serum biomarkers of prolonged erythropoiesis-stimulating agent (ESA) exposure (Darbepoietin-α) and/or aerobic training. Thirty-six healthy young males were randomly assigned to the following groups: Sedentary-placebo (n = 9), Sedentary-ESA (n = 9), Training-placebo (n = 10), or Training-ESA (n = 8). They were treated with placebo/Darbepoietin-α subcutaneously once/week for 10 weeks followed by a 3-week washout period. Training consisted of supervised biking 3/week for 13 weeks at the highest possible intensity. Serum was collected at baseline, week 3 (high dose Darbepoietin-α), week 10 (reduced dose Darbepoietin-α), and after a 3-week washout period. Serum proteins were separated according to charge and molecular mass (2D-gel electrophoresis). The identity of proteins from spots exhibiting altered intensity was determined by mass spectrometry. Six protein spots changed in response to Darbepoietin-α treatment. Comparing all 4 experimental groups, two protein spots (serotransferrin and haptoglobin/haptoglobin related protein) showed a significant response to Darbepoietin-α treatment. The haptoglobin/haptoglobin related protein spot showed a significantly lower intensity in all subjects in the training-ESA group during the treatment period and increased during the washout period. An isoform of haptoglobin/haptoglobin related protein could be a new anti-doping marker and merits further research. ClinicalTrials.gov NCT01320449.