Development and validation of an integrated tiered in vitro screening approach for predicting drug induced liver injury (DILI)

Lead Research Organisation: University of Birmingham
Department Name: Sch of Biosciences

Abstract

Liver injury is a major cause of drug attrition as a result of preclinical toxicity and usually occurs late on in clinical trials. Currently, liver injury is largely assessed using animal (rodent) models. However, there is an urgent need to develop alternative in vitro and in silico models that once validated would have a number of important potential advantages. Because multiple modes of action are known to be important an integrated tiered screening approach based on a mechanistic understanding is required. The aim of the project is o develop and assess in vitro assays including bile salt export pump and multidrug resistance protein 2 inhibition assay, UDP-glucuronosyl transferase 1A1 and 1A3 enzyme inhibition, cell toxicity (MTT, adenylate kinase), impairment of mitochondrial membrane potential (TMRE), mitochondrial (mitosox) and cellular ROS (DCF-DA) using liver cell lines (e.g. HepG2) and primary hepatocytes in combination with state-of-the-art technologies including the Hi Content Imaging Express, Incucyte Cell Imager for real time analysis and imaging of cells in culture and LC-MS-TOF for reactive metabolite assessment. In addition we will explore use of confocal-imaging in combination with tagged-reporter proteins to develop assays for stress-response mediated by specific transcriptional activation pathways (e.g. Nrf2 nuclear translocation as a marker of the "anti-oxidant response"). These assays will be validated using a "training set" of known toxic (e.g. carbon tetrachloride) and non-toxic compounds to build a database of responses to known hepatoxicants that exert their effects by different mechanisms. Once validated the predictive power of developed assays will be tested using a panel of chemicals (and metabolites, acyl,N-,O-glucuronides for example) where little information exists on their ability to cause liver injury or which are currently under in vivo assessment. A link to potential systemic drug levels will also be assessed,

Publications

10 25 50

Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/M01116X/1 01/10/2015 30/09/2023
1940003 Studentship BB/M01116X/1 02/10/2017 30/09/2021 Nicholas James Coltman
 
Description We are developing research models (multi-cellular) for use in the pharmaceutical industry that not only bypass the need for animal models, but may even be better modeling disease state, drug action/efficacy and toxicity in vivo. In drug discovery and pharmaceutical research, as well as in regulatory sciences, the use of animal models (particularly rodent models) has been mandatory policy under governing bodies such as the Food and Drug Administration (FDA, USA) and European Medicines Agency (EMA, EU) for more than 50 years, particularly in the pharmaceutical industry when novel chemical and biological entities are tested in animal models before submission to man.

With our industrial iCASE partner and academic partners at the University of Birmingham, we are developing models that can be used in both the pharmaceutical and academic environment for modeling liver biology and other useful outputs, e.g. drug metabolism and drug efficacy. We are currently assembling a manuscript where our models have been used to successfully model DNA damage and genotoxicty on exposure to common toxic agents we are exposed to in our everyday environment, such as those found in cigarette smoke for example; we therefore expect this model could be used for testing novel chemical agents also.
Exploitation Route Ultimately these models may be taken up by the pharmaceutical industry for safety assessment; alternatively these models may also serve a role in basic biological research in the academic environment.
Sectors Healthcare,Pharmaceuticals and Medical Biotechnology