OMICS-strategies and methods in the fight against doping

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Abstract

During the past decade OMICS-methods not only continued to have their impact on research strategies in life sciences and in particular molecular biology, but also started to be used for anti-doping control purposes. Research activities were mainly reasoned by the fact that several substances and methods, which were prohibited by the World Anti-Doping Agency (WADA), were or still are difficult to detect by direct methods. Transcriptomics, proteomics, and metabolomics in theory offer ideal platforms for the discovery of biomarkers for the indirect detection of the abuse of these substances and methods. Traditionally, the main focus of transcriptomics and proteomics projects has been on the prolonged detection of the misuse of human growth hormone (hGH), recombinant erythropoietin (rhEpo), and autologous blood transfusion. An additional benefit of the indirect or marker approach would also be that similarly acting substances might then be detected by a single method, without being forced to develop new direct detection methods for new but comparable prohibited substances (as has been the case, e.g. for the various forms of Epo analogs and biosimilars). While several non-OMICS-derived parameters for the indirect detection of doping are currently in use, for example the blood parameters of the hematological module of the athlete's biological passport, the outcome of most non-targeted OMICS-projects led to no direct application in routine doping control so far. The main reason is the inherent complexity of human transcriptomes, proteomes, and metabolomes and their inter-individual variability. The article reviews previous and recent research projects and their results and discusses future strategies for a more efficient application of OMICS-methods in doping control.

Introduction

In 1994 Marc Wilkins introduced the word “proteome” to the scientific community as a counterpart to the word “genome” (vide infra). A “genome” contains the entire set of genetic information of a cell or organism, while a “proteome” comprises all expressed proteins of a cell or organism [1], [2]. Subsequently, words with similar meaning were created with relation to metabolites (“metabolome”) or the RNA transcripts (“transcriptome”). The science related to these terms was named accordingly “genomics”, “transcriptomics”, “proteomics”, and “metabolomics” (Fig. 1) [3]. Within a few years an increasing number of OMICS-terms was created (e.g. glycomics, lipidomics, phosphoproteomics, neuroproteomics, degradomics, toponomics, interactomics) [4], which also was reasoned by the development of high-throughput methods for the parallel identification, characterization, and quantitation of thousands of biomolecules. OMICS-sciences have been leading to an increased understanding of diseases on the molecular level and stimulated the search for new and more specific biomarkers for the early diagnosis of diseases, their progression and prognosis, as well as the monitoring of therapies. Since similar alterations on the molecular level have been assumed for the application of prohibited doping substances, OMICS-strategies have also been applied to anti-doping research. While the ultimate goal has been the development of indirect methods for the detection of not or difficult to detect substances (e.g. hGH) and methods (autologous blood transfusion), the longitudinal monitoring of changes in “OMICS-patterns” (e.g. the pattern of endogenous steroids) added a possible new dimension to anti-doping control within the framework of the athlete's biological passport. However, currently no OMICS-derived biomarkers are used in doping testing. The main application so far has been the use of methods and instruments usually applied in proteomics (two-dimensional polyacrylamide gel-electrophoresis (2D-PAGE), or nano-liquid chromatography (LC) coupled to mass spectrometry) for the direct (targeted) detection of misused substances like peptides or hGH. In consequence, the focus of this article is on research projects, which have been using a non-targeted transcriptomic, proteomic, or metabolomic approach for the discovery of biomarkers in anti-doping control.

Section snippets

Genomics and transcriptomics

The biological information of all cellular life forms is stored in deoxyribonucleic acid (DNA) and the sequence of its nucleotides (bases). Some viruses partly use ribonucleic acid (RNA) for data-storage [2]. The entirety of this hereditary nucleic acid stored information is called a genome. Genomics is the science which deals with the exploration of genomes. The human genome consists of two parts, the nuclear genome (ca 3,200,000,000 DNA-nucleotides in size and organized in 24 chromosomes of

Proteomics

The word “proteomics” was derived from the term “proteome” in the late 1990s (vide supra) [55], [56]. Marc Wilkins introduced the word “proteome” to the scientific community in 1994 at the Siena 2D-electrophoresis meeting [57], [58]. In its original conception “proteome” meant “the PROTEins expressed by a genOME or tissue” [57] and proteome studies were primarily dedicated to the “identification and characterization of all proteins expressed by an organism or tissue” [59], [60], [61]. At that

Metabolomics

In analogy to transcriptome and proteome the term metabolome can be defined as the complete set of metabolites found in a cell, organism, or biological system [133], [134]. The term metabonomics was defined by Jeremy Nicholson in 1999 as “the quantitative measurement of the time-related multiparametric metabolic response of living systems to pathophysiological stimuli or genetic modification” [134], [135]. In 2001 the term metabolomics was defined by Oliver Fiehn as “a comprehensive and

Conclusions and future perspectives

During the past couple of years a considerable number of doping-related OMICS-projects was conducted, which was mainly driven by the poor detectability of several WADA-prohibited substances (e.g. hGH, Epo) or methods (autologous blood transfusion). The fact that the majority of these projects either failed or not directly led to applications in routine doping control is mainly due to the complexity of human transcriptomes, proteomes, and metabolomes. Additionally, the correlation between

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    This paper is part of the special issue entitled: Fight Against Doping in 2011, Guest-edited by Neil Robinson (Managing Guest Editor), Martial Saugy, Patrice Mangin, Jean-Luc Veuthey, Serge Rudaz and Jiri Dvorak.

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