Research
The CDSS Bioanalytical Facility was fundamental to research in the MRC Centre for Drug Safety Science for 10 years, providing a vital link between chemistry, biology and translational medicine, and was pivotal to the renewal of the Centre in 2014. In 2019 the Facility joined Liverpool Shared Research Facilities to widen access, ensure sustainability, and benefit from centralised administrative support. As first manager then head of the Facility, I have supported a wide range of projects, with the ultimate aim of understanding disease and effecting change in the management of patients.
Human organoid models in drug safety science (PIs Carrie Duckworth, Chris Goldring, Paul O'Neill)
Gastrointestinal (GI) toxicity is a leading cause of drug attrition. There are no robust in vitro assays or translational biomarkers that predict GI toxicity in animals or patients. We have established murine and human enteroids (complex intestinal organoids with multiple lobes and a lumen) and are investigating the interplay between the epithelium and immune system after challenge with compounds associated with severe GI adverse effects, such as irreversible tyrosine kinase inhibitors (TKIs) (Fig1). Direct TKI toxicity is believed to be a consequence of covalent binding of the drug/metabolites to proteins other than the intended targets (off-target toxicity). As well as defining the phenotypic consequences of drug exposure +/- immune mediators using SWATH, we will map the adductome of TKIs osimertinib and afatinib in human enteroids using native and biotinylated drugs, a task for which the Facility is well-qualified (20 publications on chemical modification of proteins). This work will lay the groundwork for development of the next generation of TKI, possible adjunct therapies to reduce side effects, and biomarkers to predict patient susceptibility to adverse reactions.
Drug-induced liver injury results in patient suffering and attrition of drugs. The gold standard for drug safety/toxicity assessment are human hepatocytes, but these lose crucial functions within hours under standard conditions. New culture techniques, including organoids and precison-cut liver slices, are being developed to improve the phenotypic recapitulation of the liver in vitro. SWATH/DIA analysis of these new models provides an in-depth picture of how liver-like they are and will help refine the models further. This and other work by UoL scientists has lead to a change in practice within Pharma companies.
Development of quantitative assays for monitoring changes in the activity of Nrf2 in human tissue/biofluid (PI Ian Copple)
Nrf2 is a pivotal transcription factor that orchestrates the response to oxidative stress and drives regeneration in the liver. It is gaining interest as a drug target in several disease areas, yet there are currently no validated circulating biomarkers that reflect Nrf2 modulation, hindering the ability of the field to non-invasively monitor target engagement/ pharmacodynamic responses in patients. In collaboration with industry partners, temporal blood samples are being collected from patient trials of different Nrf2 activators, and will be screened for candidate biomarkers by SWATH/DIA. This will provide supporting evidence to progress these novel drugs into the clinic and increase the likelihood of regulatory approval. There is also the potential to use Nrf2 stress response measures as a means of exploring pathways linking exposure to adverse outcomes in the context of environmental agents and medicines.
Identification of the IgG targets in autoimmune fibromyalgia (Andreas Goebel, David Andersson (UCL), Roz Jenkins)
Chronic pain disorders are the leading cause of life-years lived with disability worldwide. Fibromyalgia syndrome (FMS) affects >2% of the UK population, rising to 10-30% in patients with rheumatological conditions. Symptoms include widespread pain, fatigue and psychological distress. We demonstrated that serum-IgG from patients with severe phenotypes invariably induces FMS symptoms when injected into mice, suggesting an autoantibody-mediated mechanism. Patient IgG will be used to immunoprecipitate potential membrane-associated targets from mouse primary cells including dorsal root ganglia, and from relevant human cell lines. However, the paucity of the target proteins is a significant challenge. We are therefore developing novel applications of SWATH/DIA to enhance the identification of these difficult targets. The aim is to develop a first diagnostic serum test for this condition and also inform the development of new medications, in a field where currently few options exist.
Research grants
Development of an equine protein atlas to understand ageing and age-related diseases
HORSERACE BETTING LEVY BOARD (UK)
December 2024 - November 2027
Novel diagnostic and therapeutic insights for fibromyalgia
THE SIR JULES THORN CHARITABLE TRUST (UK)
July 2023 - June 2028
Understanding the heterogeneity of chronic lymphocytic leukaemia through the elucidation of how genetic alterations influence protein expression at the whole genome level.
NORTH WEST CANCER RESEARCH INCORPORATING CLATTERBRIDGE CANCER RESEARCH (UK)
April 2016 - March 2019
Combined Genetic and Proteomic Screening for early Detection of Pancreatic Cancer.
CANCER RESEARCH UK (UK)
October 2007 - September 2011
Regulated and deregulated secretion from primary myofibroblasts in gastric cancer.
NORTH WEST CANCER RESEARCH FUND
October 2008 - September 2011
Research collaborations
Mandy Peffers
Proteomic profiling of extracellular vesicles from equine synovial fluid and plasma to identify biomarkers of age-related conditions such as osteoarthritis
Chris Goldring
Phenotypic characterisation of novel human in vitro models using SWATH/DIA
Guozheng Wang, Simon Abrams
We are using SWATH/DIA to compare the phospho- and unmodified proteome of spleen lymphocytes from mice +/- TLR4/9 activation to determine which signalling pathways drive multi-organ failure in sepsis.
Andreas Goebel
University Hospital Aintree
Molecular mechanisms of CRPS pain
Eithne Costello
Protein expression profiling in pancreatic cancer and identification of plasma biomarkers capable of distinguishing between cancer, pancreatitis and bile duct obstruction in patients
Parveen Sharma
Mechanistic understanding of drug-induced cardiotoxicity using a novel 3D cardiomyocyte model and SWATH/DIA proteomics
Neill Liptrott
Proteomics to define inter-individual variability in immunological response to advanced therapeutics
Ian Copple
Profiling baseline and drug-stimulated hepatic proteomes to understand the translational relevance of pre-clinical models for human adverse drug reactions
Laura Watkins
Proteomic assessment of human cholangiocarcinoma liver slices as a model for screening new cancer therapies
Carrie Duckworth
Global proteomics of gastric enteroids to understand toxicity induced by therapeutic drugs
James Dear
Edinburgh University
Plasma biomarkers of drug-induced liver injury
Andrew Pettitt
Investigation of chemotherapy resistance in chronic lymphocytic leukaemia (CLL) through proteomic analysis of paired clinical trial samples obtained before treatment and at relapse. In addition, the role of CD40 stimulation in CLL survival and drug response
Ian Prior
Absolute quantification of Ras isoforms