Accelerating the Implementation of Mass Spectrometry in the Clinical Lab

CMS Logo MSACL

EUROPE 2018

Help Us Reach Our Educational Support Goal of $40,000
Educational Travel Grants supported in part by:

Plenary & Keynote Lecture Series


Distinguished Contribution Award Lecture


On the Award

>> Wednesday 16:30 in Mozart 1-5
Mass Spectrometry and the Elucidation of Steroid Metabolome
Cedric Shackleton
UCSF Benioff Children’s Hospital Oakland Research Institute, Oakland, California, and Institute of Metabolism and Systems Research (IMSR), College of Medical and Dental Sciences, University of Birmingham, UK

Sterols and steroids were among the first biomolecule groups to be studied by mass-spectrometry. The early success of GC separation in 1960 by Sweeley and Horning and the combination of GC and MS by Ragner Ryhage in Stockholm with invention of a technique for separating the bulk of carrier gas from analytes prior to ionization were fundamental. This led in the mid-60s to the first commercial dedicated GC/MS instrument, the LKB 9000. Professor Jan Sjövall (a colleague of Dr Ryhage) in particular spearheaded studies of sterols and steroids leading to multiple publications of mass spectra, and steroid quantification of serum, urinary and biliary steroids. Developments in derivatization, capillary columns and computerization were all important constituents of these developments. This presenter (a student of Sjövall) was early involved in the clinical diagnostic use of GC/MS in the 1970s and has been involved in studying urinary steroids and defining new disorders through metabolic profiles ever since.

Particle beam MS (eg. FAB) was introduced in the 80s and allowed for the first time analysis of underivatised steroids and steroid conjugates. Steroid LC/MS (Thermospray) arrived in 1987 but it was a decade later before ESI and LC/MS/MS made the use of MS in the routine steroid analytical lab a practicality. Now the technique has essentially replaced RIA in routine endocrinological applications.

GC/MS is labour-intensive and was never really suited to individual serum hormone analysis but has always been powerful for profiling urinary steroid metabolites, which now forms a branch of the recently-termed metabolome discipline. Recently there has been a re-birth in urinary steroid analysis in diagnosis and several groups are working on transferring GC/MS methodologies to LC/MS/MS to avoid need of derivatization and shorten chromatography times. This is a challenge because many of the “metabolite-type” steroids have weak ESI ionization and poor CID sensitivity. Two steroid groups are particularly suited to LC/MS/MS and those are the steroid sulfates and glucuronides since they have an existing ionic center. Analysis of intact conjugates avoids the labour-intensive enzyme hydrolysis, and we a pursuing their analysis. A drawback here is the paucity of authentic reference materials.

A huge challenge in steroid metabolomics is the presentation of diagnostic data in a form friendly to researchers and clinicians. Many recent publications have addressed this issue, including use of heat-maps and machine learning for metabolome diagnostic interpretation.

Plenary Lectures


>> Thursday 16:00 in Mozart 4-5
Mass Spectrometric Analysis of Glycomic Signatures of Cancer and Autoimmune Diseases: Towards Clinical Application
Manfred Wuhrer
LUMC

Many major human diseases including various types of cancer and autoimmune diseases are associated with protein glycosylation changes. These changes are often instrumental in disease development and progression. In cancer, glycosylation changes of both tumor tissues and tumor-derived circulating antigens have been described. However, diagnostic test often merely rely on the measurement of antigen concentrations and fail to register glycosylation changes, largely due to the lack of suitable technology and workflows. In this presentation, examples will be given of recently developed mass spectrometric workflows to determine disease-associated glycosylation changes from tumor tissues and in the circulation, but also from other body fluids including urine and saliva. Promising markers will be presented including immunoglobulin G glycosylation, cancer glycoprotein antigens, as well as tissue glycosylation signatures revealed by mass spectrometry imaging. Steps towards the clinical translation of these markers will be discussed.
>> Tuesday 14:00 in Europa
Metabolomics Messages on Human Health and Disease
Jurek Adamski
Helmholtz Zentrum München

The human metabolome represents functional read out of processes in health and disease. The metabolome is both stable and highly dynamic. Stable components are determined by genetics, genomic imprinting or physiological homeostasis. The dynamics originates from circadian rhythm, hormonal status, nutrition, environmental exposure, ageing, medication or disease. Despite its interlaced origin metabolome specifically reflects distinct processes. Metabolomics studies request therefore a special study design. Unique metabolomics signatures have been identified pre-disease (like type 2 diabetes), in disease progression (chronic kidney disease) or in companion diagnostics (drug action monitoring). Metabolomics informed diagnostics has great potential for selectivity, specificity and multiplexing of indications to be screened for.

Keynote Lectures


>> Wednesday 9:00 in Track 3 (Papageno) : Session 1
Clinical Proteomics: The Path Towards Implementation
Harald Mischak
mosaiques therapeutics GmbH & University of Glasgow

Clinical proteomics, the application of proteome analysis to clinical purpose, represents a major field in the area of proteome research. The aim of clinical proteomics is the improvement of clinical care based on (1) the identification and application of biomarkers, and (2) the suggestion of relevant therapeutic targets. These two areas have different requirements regarding specimens to be employed, technology, and data evaluation.

While substantial efforts (especially in biomarker discovery, but also in the identification of therapeutic targets) are evident based on the large number of associated publications, only a few approaches have actually resulted in clinical application. In this presentation, key issues and major challenges in clinical proteomics will be discussed. Among these are: a) the definition of a clinical need and a context-of-use, b) selection of appropriate samples, sample preparation and analytical platform, c) application of appropriate statistics, d) demonstration of benefit in a well-powered clinical study and e) obtaining regulatory approval and reimbursement, to enable actual implementation.

For several conditions and diseases, clinical proteomics has delivered solutions that are already being applied. Most successful developments are based on multi-marker panels that have demonstrated value in large studies (i.e., including at least several hundred patients). The results of these studies are expected to initiate a change in disease assessment: from diagnosis based on existing damage and therapy aiming to prevent deterioration, towards diagnosis based on molecular mechanisms, prior to observations of clinical symptoms, and therapy via correction of molecular anomalies, thereby preventing disease onset. This will also open a path towards personalized precision medicine, where intervention will be guided by molecular mechanisms, not morphological changes.

It is becoming clear that the tools required to meaningfully apply clinical proteomics (i.e., potential biomarkers, relevant technology and bio-banked samples) are available. The move from discovery towards validation and application is not only urgently necessary, but within reach. Now, a change in objective, away from additional discovery studies and towards properly testing the plethora of potential biomarkers that have been described, is needed to demonstrate the practical value of clinical proteomics.
>> Wednesday 11:00 in Track 3 (Papageno) : Session 2
Mass Spectrometry Harmonisation: Paediatric Steroid Tales
Ronda Greaves
Murdoch Children’s Research Institute

Our goal in laboratory medicine is to improve patient health by using laboratory tests as our tool to support medical decisions. The change from immunoassay to mass spectrometry analysis of steroids has significantly improved the specificity, accuracy and sensitivity of many steroids frequently analysed in children. However, mass spectrometry assays are not infallible and recognised differences exist across the total testing process. There are demonstrated variations pre-analytically, analytically and post-analytically in methods and understanding what differences are critical and how results compare between laboratories is essential to close the gap between result differences between laboratories. In this presentation, we will explore the challenges and possible solutions for the harmonisation of paediatric steroid analysis by mass spectrometry to support medical decisions.
>> Wednesday 14:30 in Track 2 (Mozart 4-5) : Session 3
Tackling Analytical Challenges in Sports Drug Testing by Mass Spectrometric Approaches
Mario Thevis
German Sport University, Cologne, Germany
European Monitoring Center for Emerging Doping Agents (EuMoCEDA), Cologne/Bonn, Germany

Sports drug testing laboratories are facing multifaceted challenges including the misuse of naturally/endogenously occurring substances, non-approved/discontinued drug candidates, urine manipulation, etc. In order to provide best-possible analytical performance, mass spectrometry-based approaches are predominantly utilized to detect prohibited substances and methods of doping. With the constantly increasing analytical requirements concerning the number of target compounds, the complexity and range of physico-chemical properties of analytes (e.g., inorganic ionic transition metals, gases, lipids, alkaloids, peptides, proteins, DNA/RNA-based drugs, etc.) as well as the desire to accelerate analyses and obtain information allowing also for retrospective data mining, high resolution/high accuracy mass spectrometry has become a mainstay in doping controls. In that context, various assays have been reported, enabling either multi-component analyses of low- or high molecular mass measurands or the specific and dedicated (confirmatory) detection of prohibited substances.

Selected applications will be presented reporting on examples of recent findings in routine sports drug testing, demonstrating both the inventiveness of cheating individuals that undermine current anti-doping efforts as well as the relevance of in-depth investigations into unusual findings, where the athletes’ innocence was to be shown albeit prohibited substances were unequivocally identified in their doping control urine samples.
>> Wednesday 14:30 in Track 4 (Paracelsus) : Session 3
Combining Mass Spectrometry Imaging & Microproteomics to Investigate Intratumor Heterogeneity
Liam McDonnell
Fondazione Pisana per la Scienza ONLUS, Pisa, Italy

Mass spectrometry imaging (MSI) is able to simultaneously record the distributions of hundreds of molecules directly from tissue. This spatially-resolved molecular information can be combined with multivariate/clustering analysis to reveal regions of tissue with distinct molecular signatures, a process that has been termed MSI-based molecular histology and has been used to reveal tumor subpopulations, metabolically distinct cell layers, and tumor interface zones. Rapid direct tissue analysis is essential for MSI in order to maintain spatial localization and acceptable measurement times. The absence of an explicit analyte separation/purification step means MSI lacks the depth of coverage of LC-MS/MS. Here, we demonstrate how MSI can be combined with high sensitivity microproteomics, even of the same tissue section, to further investigate the molecular changes associated with tumor subpopulations.
>> Thursday 9:00 in Track 5 (Doppler) : Session 4
Tandem MS-Proteotyping: Proteomics- and Genomics-Based Characterization and Typing of Infectious Bacteria
Edward Moore
University of Gothenburg, Sweden
LC-MS/MS proteomics- and genomics-based characterisation and typing of microorganisms, i.e., ‘proteotyping’, using peptides from expressed proteins, matched to genomic sequence data, can be applied for sensitive and accurate detection and typing of pathogenic bacteria. Proteotyping is capable of resolving closely-related bacterial taxa with simultaneous detections of virulence and antibiotic resistance features, providing for comprehensive characterisations of infectious bacteria. The methodology may be applied directly to analyses of clinical samples without prior cultivation and isolation, thus providing for rapid, reliable, infectious disease diagnostics.

>> Thursday 14:30 in Track 1 (Mozart 1-3) : Session 6
How Tandem Mass Spectrometry Revolutionized Newborn Screening
David Millington
Duke University - Retired

Inspired by a clinician’s account of a child rescued from near death by a revolutionary therapeutic intervention, the author applied chemistry and mass spectrometry to solve an analytical challenge that led to the first front-line diagnostic test performed by tandem mass spectrometry (MSMS) – the analysis of acylcarnitines to recognize and diagnose inherited disorders of fatty acid and branched-chain amino acid catabolism. By applying this method to dried blood spots and adding an additional analytical component to include several essential amino acids, a novel multiplex assay was developed to screen newborns for over 30 inherited metabolic conditions with a single test. The introduction of this method into public health systems and hospitals across the world during the past 20 years has literally revolutionized neonatal screening, and new technology is being introduced to add even more value to this critical “first test”. This concept subsequently became the basis of targeted metabolomics platforms that have been used, for example, to help identify new animal models of metabolic disease by screening the offspring of genetically modified adults. MSMS with UPLC has been widely applied to develop new assays for useful biomarkers of metabolic disease for both diagnosis and therapeutic monitoring. Examples from the author’s laboratory will be used to illustrate the value and scope of these methods.
>> Thursday 14:30 in Track 4 (Paracelsus) : Session 6
Advances in Computational Methods for Tissue Imaging by Mass Spectrometry
Raf Van De Plas
Delft TU
Imaging Mass Spectrometry (IMS) has made rapid progress as an imaging modality that can map the spatial distribution of molecules in tissue. In recent years, novel computational developments have become an increasingly important part of major advancements in this field. This talk presents several computational techniques developed in our group, specifically relevant to molecular imaging in medicine and the clinical practice. We show recent work in low-level signal processing, where in silico integration of isolation windows enables High-Dynamic-Range mass spectrometry, substantially increasing MS sensitivity. We also address advancements in data-driven image fusion, a multi-modal data mining methodology that drives the automated discovery of biomolecular relationships between stained microscopy and IMS, thus directly linking exploratory tissue analysis to established clinical targets.