Coupling Complete Blood Count and Steroidomics to Track Low Doses Administration of Recombinant Growth Hormone: An Anti-Doping Perspective.

Luca Narduzzi,Emmanuelle Varlet-Marie,Michel Audran,Yves Le Bouc,Marie-Line Morvan,Magnus Ericsson,Alexandre Marchand,Corinne Buisson,Bruno Le Bizec,Gaud Dervilly

doi:10.3389/fmolb.2021.683675

Abstract

Growth Hormone (GH) under its human recombinant homologue (rhGH), may be abused by athletes to take advantage of its well-known anabolic and lipolytic properties; hence it is prohibited in sports by the World Anti-Doping Agency. Due to the rapid turnover of rhGH, anti-doping screening tests have turned to monitor two endocrine biomarkers (IGF-I and P-III-NP), but unfortunately, they show population-wise variability, limiting the identification rate of rhGH users. Previous studies have evidenced the numerous effects of GH on human physiology, especially in hematopoiesis and steroidogenesis. In this work, aiming to discover novel physiological rhGH biomarkers, we analyzed the complete blood count and the steroidomics profile of healthy, physically active, young males treated either with EPO + rhGH or EPO + placebo. The time-trends of these two physiological routes have been analyzed through geometric trajectory analysis (GTA) and OPLS-DA. Individuals supplemented with micro-doses of rhGH exhibited different leukopoietic and steroidal profiles compared to the control population, suggesting a role of the rhGH in both pathways. In the article, hypotheses on the observed differences are discussed according to the most recent literature and compared to results in animal models. The use of leukopoietic and steroidal biomarkers together with endocrine biomarkers (IGF-1 and P-III-NP) allows to correctly classify over 98% of samples with no false positives, miss-classifying only one single sample (false negative) over a total of 56; a promising result, if compared to the current rhGH detection strategies.

Highlights

Growth hormone is an endocrine factor involved in the regulation of several pathways, like correct body development, metabolic homeostasis, bones and muscles anabolism
An example of the effect of Geomtetric Trajectory Analysis (GTA) on inhomogeneous groups is reported in Figure 2 with the measurements of Insulin-like Growth Factor (IGF)-I and P-III-NP, which are both biomarkers of recombinant human Growth Hormone (rhGH) administration
When original data are used, P-III-NP shows a better discrimination power than IGFI between the two groups. This happens because the concentration of IGF-I is higher in the EPO group than the EGH group before the treatments (Figure 2A); but the literature claims that IGF-I is a better marker than P-III-NP, because it is the main target of Growth Hormone (GH)’s cascade pathway and it is accumulated rapidly after rhGH treatment (Holt, 2009)

Summary

Introduction

Growth hormone is an endocrine factor involved in the regulation of several pathways, like correct body development, metabolic homeostasis, bones and muscles anabolism. Receptors for GH and IGF-I are present in several cells types of the body and they are involved in the regulation of several processes (Jones and Clemmons, 1995; Giustina and Veldhuis, 1998). Their effect is different depending on the age (Junnila et al, 2013; Blum et al, 2018), ethnicity (Berrigan et al, 2009; Grimberg et al, 2018) and sex (Meinhardt and Ho, 2006) of the subjects. Two main advantages are obtained after abusing rhGH: 1) GH exerts an anabolic effect on muscles, enhancing amino-acids uptake, limiting proteins breakdown (Bujis et al, 2002; Liu et al, 2006; Velloso, 2008). 2) GH has a lipolytic effect, driving the metabolism toward ketosis

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Conclusion