Detection of growth hormone doping by gene expression profiling of peripheral blood.
BACKGROUND
GH abuse is a significant problem in many sports, and there is currently no robust test that allows detection of doping beyond a short window after administration.


OBJECTIVE
Our objective was to evaluate gene expression profiling in peripheral blood leukocytes in-vivo as a test for GH doping in humans.


METHODS
Seven men and thirteen women were administered GH, 2 mg/d sc for 8 wk. Blood was collected at baseline and at 8 wk. RNA was extracted from the white cell fraction. Microarray analysis was undertaken using Agilent 44K G4112F arrays using a two-color design. Quantitative RT-PCR using TaqMan gene expression assays was performed for validation of selected differentially expressed genes.


RESULTS
GH induced an approximately 2-fold increase in circulating IGF-I that was maintained throughout the 8 wk of the study. GH induced significant changes in gene expression with 353 in women and 41 in men detected with a false discovery rate of less than 5%. None of the differentially expressed genes were common between men and women. The maximal changes were a doubling for up-regulated or halving for down-regulated genes, similar in magnitude to the variation between individuals. Quantitative RT-PCR for seven target genes showed good concordance between microarray and quantitative PCR data in women but not in men.


CONCLUSIONS
Gene expression analysis of peripheral blood leukocytes is unlikely to be a viable approach for the detection of GH doping.
Mitchell
Christopher J
CJ
Pituitary Research Unit, Garvan Institute of Medical Research, 384 Victoria Street, Darlinghurst New South Wales 2010, Sydney, Australia.
Nelson
Anne E
AE
Cowley
Mark J
MJ
Kaplan
Warren
W
Stone
Glenn
G
Sutton
Selina K
SK
Lau
Amie
A
Lee
Carol M Y
CM
Ho
Ken K Y
KK
D000328:Adult; D004300:Doping in Sports / Q000379:methods*; D004311:Double-Blind Method; D005260:Female; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D019382:Human Growth Hormone / Q000494:pharmacology*; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012727:Sex Characteristics; D055815:Young Adult
D011994:Recombinant Proteins; D019382:Human Growth Hormone; D012313:RNA; D007334:Insulin-Like Growth Factor I