WADA Implications of RNA-seq in the detection of anabolic steroid use and the harnessing of the molecular mechanism(s) of muscle memory

This project aimed to determine the histological and morphological changes within human skeletal muscles as a result of the administration of AAS.

The effects of Anabolic Androgenic Steroids (AAS) are largely illustrated through Androgen Receptor induced gene transcription, yet RNA-Seq has yet to be conducted on human whole blood and skeletal muscle. Investigating the transcriptional signature of AAS in blood may aid AAS detection and in muscle further understanding of AAS induced hypertrophy.

Aims:
To establish skeletal muscle gene markers (in the first instance) elicited by AAS.
To integrate the molecular, histological and training response data to help elucidate the short- and long-term effects of AAS and implications for anti-doping. To refine the detection of AAS

Cross-comparison of both standard reagent whole blood (N = 55: C = 7, RT = 20, RT-AS ≤ 2 = 14, RT-AS ≥ 10 = 10, RP = 4; N = 46: C = 6, RT = 17, RT-AS ≤ 2 = 12, RT-AS ≥ 10 = 8, RP = 3) sequencing datasets, showed that no genes or gene sets/pathways were differentially expressed between time points for RP or between group comparisons of RT-AS ≤ 2 vs. C, RT, or RT-AS ≥ 10. Cross-comparison of both muscle (N = 51, C = 5, RT = 17, RT-AS ≤ 2 = 15, RT-AS ≥ 10 = 11, RP = 3) sequencing (one standard & one CoolMPS reagent) datasets, showed one gene, CHRDL1, which has atrophying potential, was upregulated in RP visit two. In both muscle sequencing datasets, nine differentially expressed genes, overlapped with RT-AS ≤ 2 vs. RT and RT-AS ≤ 2 vs. C, but were not differentially expressed with RT vs. C, possibly suggesting they are from acute doping alone. No genes seemed to be differentially expressed in muscle after the long-term cessation of AAS, whereas a previous study found long term proteomic changes.

Conclusion:

A whole blood transcriptional signature of AAS doping was not identified. However, RNA-Seq of muscle has identified numerous differentially expressed genes with known impacts on hypertrophic processes that may further our understanding on AAS induced hypertrophy. Differences in training regimens in participant groupings may have influenced results. Future studies should focus on longitudinal sampling pre, during and post-AAS exposure to better control for confounding variables.

Primary publication:
An observational human study investigating the effect of anabolic androgenic steroid use on the transcriptome of skeletal muscle and whole blood using RNA-Sequencing. Kolliari-Turner, A., Lima, G., Wang, G., Malinsky, F. R., Karanikolou, A., Eichhorn, G., Tanisawa, K., Ospina-Betancurt, J., Hamilton, B., Kumi, P. Y. O., Shurlock, J., Skiadas, V., Twycross-Lewis, R., Kilduff, L., Martin, R. P., Ash, G. I., Potter, C., Guppy, F. M., Seto, J. T. & Fossati, C. & 3 others, , 3 May 2023, In: BMC Medical Genomics. 16, 1, 16 p., 94.