Dr Roz Jenkins BSc (Hons), PhD

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.

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.

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.