Transcriptomic Markers of Recombinant Human Erythropoietin Micro-Dosing in Thoroughbred Horses

Recombinant human erythropoietin (rHuEPO) is a well-known performance enhancing drug in human athletes, and there is anecdotal evidence of it being used in horse racing for the same purpose. rHuEPO, like endogenous EPO, increases arterial oxygen content and thus aerobic power. Micro-doping, or injecting smaller doses over a longer period of time, has become an important concern in both human and equine athletics since it is more difficult to detect. Horses offer an additional challenge of a contractile spleen, thus large changes in the red blood cell mass occur naturally. To address the challenge of detecting rHuEPO doping in horse racing, we determined the transcriptomic effects of rHuEPO micro-dosing over seven weeks in exercised Thoroughbreds. RNA-sequencing of peripheral blood mononuclear cells isolated at several time points throughout the study identified three transcripts (C13H16orf54, PUM2 and CHTOP) that were significantly (PFDR < 0.05) different between the treatment groups across two or three time point comparisons. PUM2 and CHTOP play a role in erythropoiesis while not much is known about C13H16orf54, but it is primarily expressed in whole blood. However, gene expression differences were not large enough to detect via RT-qPCR, thereby precluding their utility as biomarkers of micro-doping.

Cross-platform transcriptomic profiling of the response to recombinant human erythropoietin

RNA-seq has matured and become an important tool for studying RNA biology. Here we compared two RNA-seq (MGI DNBSEQ and Illumina NextSeq 500) and two microarray platforms (GeneChip Human Transcriptome Array 2.0 and Illumina Expression BeadChip) in healthy individuals administered recombinant human erythropoietin for transcriptome-wide quantification of differential gene expression. The results show that total RNA DNB-seq generated a multitude of target genes compared to other platforms. Pathway enrichment analyses revealed genes correlate to not only erythropoiesis and oxygen transport but also a wide range of other functions, such as tissue protection and immune regulation. This study provides a knowledge base of genes relevant to EPO biology through cross-platform comparisons and validation.

Evaluation of Recombinant Human Erythropoietin Responsiveness by Erythrocyte Creatine in Haemodialysis Patients

Background: One of the main causes of anaemia in patients with end-stage renal disease is relative deficiency in erythropoietin production. Recombinant human erythropoietin (rHuEpo), a potent haematopoietic growth factor, is used to treat anaemia in haemodialysis patients. The effect of rHuEpo is usually assessed by haematological indices such as red blood cell count, haemoglobin concentration and haematocrit, but erythrocyte indices do not provide information of the rapid change in erythropoietic activity. As erythrocyte creatine directly assess erythropoiesis, the aim of this study was to evaluate the effect of rHuEpo in haemodialysis patients by measuring erythrocyte creatine. Methods: rHuEpo dose was fixed 3 months prior to the enrollment and was maintained throughout the entire study period. Eerythrocyte creatine was measured with haematologic indices in 83 haemodialysis patients. Haemoglobin was also measured 3 months after. Results: rHuEpo dose (152.4±62.9 vs. 82.2±45.5 units/kg/week, P=0.0001) and erythrocyte creatine (2.07±0.73 vs. 1.60±0.41 µmol/gHb, p=0.0003) were significantly higher in 27 patients with haemoglobin <10g/dL compared to 56 patients with haemoglobin ≥10g/dL. There was a fair correlation between rHuEpo dose and erythrocyte creatine (r=0.55, P <0.0001). Increase in haemoglobin (>0.1g/dL) was observed in 37 patients, whereas haemoglobin did not increase in 46 patients. Erythrocyte creatine was significantly higher in patients with increase in haemoglobin compared to those without (2.04±0.64 vs. 1.52±0.39 µmol/gHb, p <0.0001). When 8 variables (rHuEpo dose, erythropoietin resistance index, C-reactive protein, intact parathyroid hormone, incidence of iron deficiency, presence of anaemia, erythrocyte creatine and reticulocyte) were used in the multivariate logistic analysis, erythrocyte creatine emerged as the most important variable associated with increase in haemoglobin (Chi-square=6.19, P=0.01). Conclusion: Erythrocyte creatine, a useful marker of erythropoietic capacity, is a reliable marker to estimate ameliorative effectiveness of rHuEpo in haemodialysis patients.

Roxadustat Improves Erythropoietin Antibody-Mediated Pure Red Cell Aplasia in a Patient with Hemodialysis

Anemia is a common complication of chronic kidney disease (CKD). Recombinant human erythropoietin (rHu-EPO) is used extensively in patients with CKD. However, anti-erythropoietin (anti-EPO) antibody has been reported during rHu-EPO treatment, which causes pure red cell aplasia (PRCA). We presented a case of 75-year-old man, who underwent hemodialysis for 2 years. He developed PRCA during rHu-EPO treatment. The rHu-EPO was immediately discontinued, and the patient was given roxadustat treatment. After 6 months of roxadustat treatment, the anti-EPO antibody was disappeared, and hemoglobin recovered normal range. The results suggest that roxadustat can be used to treat patients with anti-EPO antibody-mediated PRCA without immunosuppressive therapy.

Preoperative Preparation of Patients with Anemia before Shoulder Replacement

The preoperative anemia in patients increases the risk of perioperative blood transfusion, myocardial infarction, ischemic stroke, acute kidney injury, and higher nosocomial and 30-day mortality, lengthens the duration of treatment, and increases the risk of re-hospitalization.The objective: to improve treatment results of patients after revision shoulder replacement through management of preoperative anemia.Subjects and methods. 170 medical files of patients who underwent revision shoulder replacement from 2014 to 2021 were retrospectively analyzed. On the day when they were examined by the physician, all patients had blood hemoglobin level below 130 g/l. The patients were divided into two groups that were comparable in their characteristics. Patients of the Main Group (n = 90) received preoperative preparation with iron carboxymaltate in combination with recombinant human erythropoietin 4 weeks before the expected date of surgery. Patients of the Control Group (n = 80) did not have any specific preparation.Results. The data obtained showed that the volume of intraoperative and postoperative drainage blood loss did not differ statistically between the groups; hemoglobin blood level in patients from the Main Group was statistically significantly higher both before the surgery and on the first day after it, and no hemotransfusion was indicated. In the Control Group, a statistically significantly greater decrease in hemoglobin level was observed in the postoperative period; 3 (3.8%) patients in the Control Group required RBC-transfusion. Complications in the early postoperative period (myocardial infarction, pulmonary embolism, and cardiac arrhythmia) were significantly less frequent in the Main Group versus the Control one (4.5% of cases vs. 8.8%; p < 0.05).Conclusion. The use of iron carboxymaltate at the dose of 1,000 mg once and, if necessary, repeatedly in 14 days in combination with recombinant human erythropoietin at the dose of 150 IU/kg allows preparing patients for revision shoulder replacement within the period from two weeks to one month. Preoperative management of anemia until the hemoglobin concentration reaches 130 g/l using the proposed regimen can significantly reduce the need for blood transfusion and frequency of somatic complications in the early postoperative period.