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This version published online on October 29, 2009
Journal of Clinical Endocrinology & Metabolism , doi:10.1210/jc.2009-1038
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Submitted on May 15, 2009
Accepted on September 18, 2009

Detection of Growth Hormone Doping by Gene Expression Profiling of Peripheral Blood

Christopher J. Mitchell, Anne E. Nelson, Mark J. Cowley, Warren Kaplan, Glenn Stone, Selina K. Sutton, Amie Lau, Carol M. Y. Lee, and Ken K. Y. Ho*

Pituitary Research Unit (C.J.M., A.E.N., S.K.S., A.L., C.M.Y.L., K.K.Y.H.) and Peter Wills Bioinformatics Centre (M.J.C., W.K.), Garvan Institute of Medical Research; St. Vincent's Hospital (K.K.Y.H.) and St. Vincent's Clinical School, University of New South Wales (K.K.Y.H); Sydney 2010, and Commonwealth Scientific and Industrial Research Organization Mathematical and Information Sciences, North Ryde (G.S.), Sydney, New South Wales 2113, Australia

* To whom correspondence should be addressed. E-mail: k.ho{at}garvan.org.au.

Context: 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.

Design: 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.

Conclusion: Gene expression analysis of peripheral blood leukocytes is unlikely to be a viable approach for the detection of GH doping.






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