Improving health outcomes of transplant patients

Stephen Bustin

Professor Stephen Bustin is Professor of Molecular Medicine at our Postgraduate Medical Institute. For 30 years he has investigated the biology of cancers, focusing on the role of endocrine factors on tumourigenesis.

Many of Stephen's publications describe important advances in colorectal and breast cancers, and a continuing research theme has been translating novel ideas into clinical practice. He holds editorial and committee positions in his field and is Editor-in-Chief of the journal Biomolecular Detection and Quantification.

After identifying widespread concerns about published research results, Stephen is working on developing a new technique for accurately identifying pathogens and is aiming to establish a centre of excellence for molecular diagnostics.

How did the work come about?

I have a long-standing interest in applying molecular technologies to translational research. The Polymerase Chain Reaction (PCR) is the most common molecular technique in use today and I have been working with it for many years. A review I published in 2000 has become the most cited on this technique (over 3,000 citations) and I have written and edited three books on this topic. I realised early on that there

were real problems with how researchers carry out their experiments. I have spent the last 16 years identifying the pitfalls associated with this technique and suggesting solutions.

What is the impetus for establishing the centre?

Molecular diagnostics refers to the detection of cellular disease associated biomarkers or the identification of pathogens using antibodies or the PCR. Cellular biomarkers comprise genetic and epigenetic alterations of DNA or RNA expression signatures and are used for the diagnosis and prognosis of cancers. Unfortunately, the reliability of many of these biomarkers is open to serious doubt. Last year, we published in Nature Methods a survey of 2,000 peer-reviewed publications that use PCR and showed that virtually every paper we looked at was seriously flawed. We followed this up with a paper published earlier this year that looked specifically at

colorectal cancer. It concluded that most prognostic biomarkers for this cancer are unlikely to be clinically useful. A second interest involves developing new approaches to improve the speed and accuracy of detecting viable and infectious fungal and bacterial pathogens.

Why focus on fungal pathogen detection?

The impaired immune system of immunosuppressed transplant patients makes any infection life threatening and so early and reliable detection of pathogens is particularly important. The problem is that, although fungi are ubiquitous, their accurate identification as infectious agents is rather challenging. The current approach (using clinicians’ gut instinct to decide on the correct treatment) often leads to the administration of inappropriate antibiotics and wastes millions of pounds a year.

What will this work lead to?

A better test that will save lives, we hope. The problem with current antibody-based diagnostic tests is that, while they can identify viable pathogens, they’re not sufficiently sensitive for earliest possible diagnosis. PCR assays, on the other hand, are extremely sensitive, but provide no information of the infectivity of the bug they’re detecting. Our approach combines the advantages of the two methods in an assay developed by US biotechnology company, Life Technologies, called a Proximity Ligation assay. It allows us to use a quantitative PCR assay to detect the binding of an antibody that specifically targets proteins expressed only by actively growing fungi. This should allow us to detect the presence of the infectious agent up to a week earlier than is currently possible with antibody-based tests. Consequently, antibiotics administration should become more targeted, saving money and potentially lives.

Have there been any outcomes so far?

Since my arrival at Anglia Ruskin in September 2012, we’ve published 12 peer reviewed papers and several book chapters, and one of the assays we helped develop is in regular use with several NHS Trusts. We’re regularly invited to give talks around the world and I’ve just returned from two lecture tours (one of the USA, the other of India, Malaysia, Indonesia and Australia). We also received a grant from the European Molecular Biology Organisation (EMBO) to run an international workshop on the Proximity Ligation assay in Chelmsford and are recognised worldwide as being at the very forefront in our particular area.

Research Matters

Your inspiration?

My inspiration is to do things that are useful and have a positive impact on people’s lives. I like to work in areas where I can see some kind of impact or return within a reasonable period of time, rather than just addressing questions of scientific interest.

Any surprises?

The poor quality of a lot of the research in the field, how the results are over interpreted and how much contradiction there is in the results published.

Why does this research matter?

The publication of erroneous results that lead to wrong conclusions can negatively affect people’s lives, as we have seen with the MMR/autism farce. This has caused a great deal of unhappiness and even deaths. So it matters very much.

Research funding

Life Technologies, San Francisco, CA

Amount of funding awarded: $50,000

OLM Medical, Newcastle

Amount of funding awarded: £66,000

Reading matter

Real-time quantitative PCR, pathogen detection and MIQE

Methods Mol Biol. 2013 PMID: 23104279

Johnson G, Nolan T, Bustin S.A.

RNA biomarkers in colorectal cancer

Methods. 2012 PMID: 23079397

Bustin S.A., Murphy J

A MIQE-compliant real-time PCR assay for Aspergillusdetection

PLoS One. 2012 PMID: 22808087

Johnson G.L., Bibby D.F., Wong S, Agrawal S.G., Bustin S.A., Agrawal S.G.