Analysis of human reticulocyte genes reveals altered erythropoiesis: potential use to detect recombinant human erythropoietin doping.
OBJECTIVE
Enhancement of oxygen delivery to tissues is associated with improved sporting performance. One way of enhancing oxygen delivery is to take recombinant human erythropoietin (rHuEpo), which is an unethical and potentially dangerous practice. However, detection of the use of rHuEpo remains difficult in situations such as: i) several days after the end of treatment ii) when a treatment with low doses is conducted iii) if the rHuEpo effect is increased by other substances. In an attempt to detect rHuEpo abuse, we selected erythroid gene markers from a SAGE library and analyzed the effects of rHuEpo administration on expression of the HBB, FTL and OAZ genes.


METHODS
Ten athletes were assigned to the rHuEpo or placebo group. The rHuEpo group received subcutaneous injections of rHuEpo (50 UI/kg three times a week, 4 weeks; 20 UI/kg three times a week, 2 weeks). HBB, FTL and OAZ gene profiles were monitored by real time-polymerase chain reaction (PCR) quantification during and for 3 weeks after drug administration.


RESULTS
The global analysis of these targeted genes detected in whole blood samples showed a characteristic profile of subjects misusing rHuEpo with a increase above the threshold levels. The individual analysis of OAZ mRNA seemed indicative of rHuEpo treatment.


CONCLUSIONS
The performance-enhancing effect of rHuEpo treatment is greater than the duration of hematologic changes associated with rHuEpo misuse. Although direct electrophoretic methods to detect rHuEpo have been developed, recombinant isoforms of rHuEpo are not detectable some days after the last subcutaneous injection. To overcome these limitations indirect OFF models have been developed. Our data suggest that, in the near future, it will be possible to consolidate results achievable with the OFF models by analyzing selected erythroid gene markers as a supplement to indirect methods.
Varlet-Marie
Emmanuelle
E
Biophysical & Bioanalysis Laboratory, Faculty of Pharmacy, University Montpellier I, Montpellier, France.
Audran
Michel
M
Lejeune
Mireille
M
Bonafoux
Béatrice
B
Sicart
Marie-Therese
MT
Marti
Jacques
J
Piquemal
David
D
Commes
Thérèse
T
D000293:Adolescent; D000328:Adult; D015415:Biomarkers / Q000097:blood; D004300:Doping in Sports; D004920:Erythropoiesis / Q000235:genetics*; D004921:Erythropoietin / Q000097:blood*; D005786:Gene Expression Regulation; D006801:Humans; D010919:Placebos; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins; D012156:Reticulocytes / Q000502:physiology*; D013177:Sports
D015415:Biomarkers; D010919:Placebos; D012333:RNA, Messenger; D011994:Recombinant Proteins; D004921:Erythropoietin