Drug Monitoring and Metabolism Research: Highlights from Viapath’s Fourth Innovation Academy

A host of dynamic and influential scientists, lecturers and clinicians in the pathology field convened in London to present important scientific findings for the fourth Innovation Academy Scientific Symposium.

The Grocers’ Hall in the City of London was the opulent setting for the fourth Innovation Academy (IA4) on December 5, 2014. The Innovation Academy, organised by Viapath, meets twice yearly, bringing together world-renowned leaders in pathology and innovative young clinical scientists, with this session focusing on Next Generation Diagnostics. Here we examine the effect of next generation innovations on drug monitoring and metabolism research by profiling the presentations from two of the Academy’s impressive line-up of speakers.

Doping and Detection of Anabolic Steroids

One of the guest speakers was Dr Andrew Kicman, Head of Research and Development at the Drug Control Centre, Department of Forensic Science and Drug Monitoring at King’s College London. Andrew gave a fascinating talk about Anabolic Steroids in the Gym; Doping and Detection. The use of drugs to enhance performance is considered unethical by most international sports organisations as well as the public; and the majority wish to keep sport clean. Unfortunately, doping continues to be a problem, with the World Anti-Doping Accredited (WADA) laboratories reporting 2% adverse or atypical (drug- or stimulant-positive) findings from nearly 270,000 samples tested worldwide in 2013 (the latest statistics available). Year on year, anabolic steroids account for the majority of these findings.

In society, a British Crime Survey (published in 2010) reported that the prevalence of anabolic steroid use was similar in number to use of heroin or crack cocaine (50,000 users). Moreover, reports suggest that anabolic steroids are now one of the commoner forms of injected drugs (results gleaned from needle exchange programme data)[i]. Anabolic steroids detected in dietary supplements for bodybuilding also pose a significant risk to public health, in particular among teenage boys keen to build muscle mass. For example, in an analysis of 24 bodybuilding products sold online and from UK fitness shops, 23 of them contained steroids including known anabolic agents, 16 contained mislabelled steroids – different to those indicated on the packaging, including potentially hepatotoxic steroids at doses that could present a genuine health risk[ii].

A further area of concern is with stimulants such as DMAA (1,3-dimethylamylamine or methylhexanamine), originally marketed as a (now withdrawn) nasal decongestant. Such stimulants are extensively marketed as energy-boosting, weight-loss and pre-workout dietary supplements. However, several adverse events and at least three deaths have been associated with DMAA-containing supplements[iii], including that of a marathon runner[iv]. Recently, the supply of DMAA has been curtailed because of action by the US Food and Drug Administration, but other aliphatic amines have become available, such as DMBA.

Readily Available

Despite many anabolic steroids being withdrawn as licensed products in the UK and several other countries from the 1980s onwards, there is still a readily-available source worldwide for non-medicinal purposes. Of note is that the clinical usefulness of anabolic steroids is currently being revisited, such as in the treatment of AIDS-associated cachexia, severe burns, renal failure and sarcopenia in the elderly. Even with steroids that have an enhanced anabolic effect, e.g. nandrolone and stanozolol, their androgenic effects cannot be divorced entirely with their use (such as increased cardiovascular risk factors, gynaecomastea, hirsutism and virilisation, amenorrhea and infertility in women, testicular atrophy in males, acne, mood disturbances and increased aggression).

In spite of the unfavourable anabolic-androgenic index of testosterone compared to many of its chemical analogues, in sport, testosterone continues to rank as number one in the findings of the class of anabolic agents. This is because it is tricky to prove a doping offence, as it is also produced naturally within the body. The standard approach measures the urinary testosterone to epitestosterone (T/E) ratio, which includes a screen based on a Bayesian statistical approach, where each individual acts as his/her own reference (known as the Athlete Biological Passport). Further confirmation can be made using carbon isotope mass spectrometry to discern whether steroids are of pharmaceutical or endogenous origin.

Dr Kicman concluded: “Unfortunately, users can still evade detection by the co-administration of epitestosterone with testosterone, which can normalize the T/E ratio. This makes a case for adopting a multifaceted approach in detection of testosterone doping, including the longitudinal profiling of testosterone in serum. This approach was taken in a pilot collaborative study between King’s College London and UK Anti-Doping (UKAD) regarding serum T profiling which has been completed and the results were encouraging.”

Following his presentation, Robert Dunn, a member the Future Leaders in Innovation Group at Viapath, discussed the detection of doping in sport with Dr Kicman and asked him about his long-standing career in toxicology. Read the interview here.

PCOS-Related Changes in Serum Steroid Concentrations

Lea Ghataore, Viapath Chief Biomedical Scientist, deputy in the Steroid Laboratory at King’s College Hospital provided the attendees with an overview of polycystic ovarian syndrome (PCOS), a heterogeneous disorder affecting up to 20% of women of reproductive age. As hyperandrogenism plays a central role in the pathophysiology of PCOS and is associated by factors such as insulin resistance and obesity, the Innovation Academy-funded project assessed the effect of insulin resistance and fat distribution on steroid metabolism.

The cross-sectional study, from Plymouth Hospital, UK, recruited 33 PCOS women (14 lean and 19 obese) and 30 control women (16 lean and 14 obese), matched for age and body mass index (BMI). Fat distribution was assessed by waist circumstance, skin fold measurements and body mass index. Insulin resistance was measured using Homeostatic Model Assessment (HOMA-IR). Subjects underwent glucose tolerance test (GTT) after an overnight fast and blood samples were collected at baseline, 15, 30, 60, 90 and 120 min and a serum steroid panel (androstenedione, testosterone, 17oh-pregnenolone, 17oh-progesterone, dehydroepiandrosterone sulphate, corticosterone, cortisol & cortisone), was measured by an in-house method using LC-MS/MS (TSQ Vantage, ThermoFisher Scientific).

Baseline testosterone, corticosterone and 17oh-pregnenolone were higher in the PCOS group vs. the control group. Following OGTT, attenuated responses were found for androstenedione and cortisol in the PCOS group. HOMA-IR was similar between both groups; however, the obese individuals in both groups had higher HOMA-IR. No correlations were found between the steroids measured and fat distribution/insulin resistance.

The changes in steroid metabolism in our cohort were more likely due to PCOS rather than fat distribution/insulin resistance. Higher 17oh-pregnenolone in PCOS has not been previously reported. It remains to be elucidated if the altered steroid metabolism is a contributing factor to the pathogenesis or a consequence of PCOS. A serum ‘steroid panel’ provides advantages over single steroid measurements in excluding major causes of androgen excess.

Dominic Harrington, Scientific Director at Viapath commented: “The delivery of Next Generation Diagnostics is about more than a ground-breaking technology, smart techniques, simple tests or therapies. To foster its true potential requires the alignment of multiple diagnostics, devices, IT, educational platforms and professional networks. Viapath’s Innovation Academy drives inter-disciplinary communication and helps build such networks. We are grateful for all the speakers and attendees for their fascinating contributions in making December’s IA4 a genuine forum for change.”

References

[i] ACMD Advisory Council on the Misuse of Drugs, Consideration of the Anabolic Steroids, September 2010. Available from: https://www.gov.uk/government/uploads/system/uploads/attachment_data/file/119132/anabolic-steroids.pdf

[ii] Abbate V et al. Anabolic steroids detected in bodybuilding dietary supplements - a significant risk to public health. Drug Testing and Analysis (Wiley).Online Early View (2014) available from: https://archive.org/stream/pdfy-ezHlaJn8SabPRJkO/Anabolic%20steroids%20in%20supplements_djvu.txt

[iii] Singer, Natasha; Peter Lattman (April 16, 2013). "F.D.A. Issues Warning on Workout Supplement". New York Times. Retrieved April 16, 2013.

[iv] Archer et al, Running an unknown risk: a marathon death associated with the use of 1,3 dimethylamylamine (DMAA). Drug Testing and Analysis (Wiley).Online Early View (2015) available from: http://onlinelibrary.wiley.com/doi/10.1002/dta.1764/pdf