Proteomic Biomarker Research and Systems Medicine
The Biomarkers and Systems Medicine (BSM) group was set up by Professor Harald Mischak to investigate the use of proteomic/peptidomic biomarkers as a method for the early detection of disease states. He has been joined in his research by Dr Bill Mullen whose previous focus was in metabolomics in nutrition and health.
The aim of the group was to develop panels of biomarker from body fluids that would provide clinically applicable tests in the diagnosis and monitoring of diseases, as well as being able to provide clinicians details of progress in therapeutic treatments. The long term goal being personalised medicine, where simple analysis of an individual’s proteome can help diagnose specific conditions and inform and monitor treatment.
Conventional biomarkers have failed to provide sufficient sensitivity and selectivity to allow early detection of many diseases (Figure 1).
The BSM group, in conjunction with several leading research institutions worldwide (link 3), have developed urinary tests that detect the onset of a range of diseases, before the occurrence of classic symptoms; these include coronary artery disease (CAD), preeclampsia, chronic kidney disease (CKD), diabetic nephropathy (DN), prostate and bladder cancer (Figure 2).
The non-invasive test, based on the analysis of the urinary proteome, can assist in the early diagnosis of a number of diseases, in monitoring their progress and in the efficacy of their treatment. In addition, the identification of the individual disease biomarkers and the proteins that they originate from provides an insight to the pathophysiological changes that occur at the onset of the disease. The researchers aim to utilize this knowledge in the development of novel drug candidates
The group uses CE-MS technology to display peptides and proteins of body fluids. The generated pattern, filed in a database, provides a fast and authentic description of the individual health of an organism. Measurement of the proteome in combination with on-line database matching is the key to an all-in-one diagnosis for a wide spectrum of clinical applications (Figure 3).
The overall principle of this ground-breaking technology is the fact that proteins are indispensable for the function of cells and organs, as well as for all communication inside the body. They also catalyze chemical reactions and protect the body against pathogens. Thousands of proteins and peptides are involved in these processes, forming a distinct pattern, which changes with disease. The database can identify up to 6,000 of such proteins and polypeptides and subsequently generates disease specific patterns. This vast wealth of information facilitates accurate diagnosis of diseases and enables effective therapy prior to serious organ damage.
Studying proteins rather than genes is important as can be seen by looking at the life cycle of a butterfly. As the butterfly develops from caterpillar to butterfly its genome does not change but its proteome does, as different proteins are expressed to produce very different creatures at different times (Figure 4).
One of the major challenges in proteomics is the accurate display of the peptides and proteins of body fluids, providing an opportunity to get a fast and authentic description of the individual health of an organism. Measurement of the proteome in combination with on-line database matching is the key to an all-in-one diagnosis for a wide spectrum of clinical applications (Figure 5).
Once these biomarker fingerprints have been validated it is possible to then sequence the peptides that make up the patterns. The group has the latest state of the art Orbitrap Velos mass spectrometer with nanoflow HPLC system to sequence the individual peptides. Once these peptides have been sequenced the originating proteins can be identified (Figure 6). This provides an insight to the pathophysiological changes that occur at the onset of the disease. The researchers aim to utilize this knowledge in the development of novel drug candidates.
The biomarkers lab specialises in the identification of peptides and proteins in body fluids (mostly urine) that correlate with disease, hence can be used as biomarkers in the diagnosis of disease, assessment of therapeutic intervention, and also establishment of molecular biology. Using multiparametric algorithms, the biomarkers can be combined to classifiers for a variety of pathologies. The group has pioneered clinical proteomics, established guidelines for biomarker identification and qualification, and is renowned as one of the leading groups in this area worldwide. Various diagnostic tests based on the technology have been developed, and some of these are already being used for clinical evaluation of patients. Several biomarkers have been filed with the regulatory agencies, FDA and EMA, for qualification as biomarkers in various disease conditions.
- Carty DM, Siwy J, Brennand JE, Zurbig P, Mullen W, Franke J, McCulloch JW, North RA, Chappell LC, Mischak H, Poston L, Dominiczak AF, Delles C. Urinary Proteomics for Prediction of Preeclampsia. Hypertension 2011;57(3):561-9.
- Lopez LM, Mullen W, Zurbig P, Harris SE, Gow AJ, Starr JM, Porteous DJ, Mischak H, Deary IJ. A pilot study of urinary peptides as biomarkers for intelligence in old age. Intelligence 2011;39:46-53.
- Albalat A, Mischak H, Mullen W. Urine proteomics in clinical applications: technologies, principal considerations and clinical implementation. Prilozi 2011;32:13-44.
- Kistler AD, Siwy J, Breunig F, Jeevaratnam P, Scherl A, Mullen W, Warnock DG, Wanner C, Hughes DA, Mischak H, Wüthrich RP, Serra AL. A distinct urinary biomarker pattern characteristic of female Fabry patients that mirrors response to Enzyme Replacement Therapy. PloS ONE 2011;6:e20534.
- Metzger J, Chatzikyrkou C, Broecker V, Schiffer E, Jaensch L, Iphoefer A, Mengel M, Mullen W, Mischak H, Haller H, Gwinner W. Diagnosis of subclinical and clinical acute T-cell mediated rejection in renal transplant patients by urinary proteome analysis. Proteomics Clin Appl 2011;5:322-33.
- Siwy J, Mullen W, Golovko I, Franke J, Zurbig P. Human urinary peptide database for multiple disease biomarker discovery. Proteomics Clin Appl 2011;5:367-74.
- Mullen W, Gonzalez J, Siwy J, Franke J, Sattar N, Mullan A, Roberts S, Delles C, Mischak H, Albalat A. The effect of short-term consumption of a polyphenol rich drink on biomarkers of coronary artery disease defined by urinary proteomics J Agr Food Chem 2011;59:12850-7.
- Mullen W, Albalat A, Gonzalez J, Zerefos P, Siwy J, Franke J, Mischak H. Performance of different separation methods interfaced in the same MS-RTOF detector, a comparison of performance between Capillary Electrophesis versus High performance Liquid Chromatography for biomarker analysis Electrophoresis 2012;33:567-74.
- Mullen W, Delles C, Mischak H. Urinary proteomics in the assessment of chronic kidney disease. Curr Opin Nephrol Hypertens 2011;20:654-61.
- Kitson SM, Mullen W, Cogdell RJ, Bill RM, Fraser NJ. GPCR production in a novel yeast strain that makes cholesterol-like sterols. Methods 2011;55:287-92.
- Albalat A, Mischak H, Mullen W. Clinical application of urinary proteomics/peptidomics. Expert Rev Proteomics 2011;8:615-29.
- Jahn H, Wittke S, Zurbig P, Raedler TJ, Arlt S, Kellmann M, Mullen W, Eichenlaub M, Mischak H, Wiedemann K. Peptide fingerprinting of Alzheimer's disease in cerebrospinal fluid: identification and prospective evaluation of new synaptic biomarkers. Plos One 2011;6:e26540.
- Dawson J, Walters MR, Delles C, Mischak H, Mullen W. Urinary Proteomics to Support Diagnosis of Stroke. PLoS One 2012;7:e35879.
- Siwy J, Metzger J, Hanig JP, Rodney R, Mullen W, Mischak H. Proteomic Biomarkers of Drug-Induced Nephrotoxicity in the Rat. Plos One 2012;7:e34606.
- Kuznetsova T, Mischak H, Mullen W, Staessen JA. Urinary proteome analysis in hypertensive subjects with left ventricular diastolic dysfunction. European Heart Journal 2012
- Metzger J, Negm AA, Plentz RR, Weismüller TJ, Wedemeyer J, Karlsen TH, Mullen W, Mischak H, Manns MP, Lankisch TO. Urine proteomic analysis differentiates cholangiocarcinoma from primary sclerosing cholangitis and other benign biliary disorders, Gut 2012.
- Rouse R, Siwy J, Mullen W, Mischak H, Metzger J, Hanig J. Proteomic Candidate Biomarkers of Drug-Induced Nephrotoxicity in the Rat. Plos One 2012