PhD Research Project: Development and validation of an integrated tiered in vitro screening approac
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 including:
1) They would be simpler and cheaper.
2) They would be amenable to high throughput screening.
3) They could be designed based on mechanistic understanding of liver injury to test for different classes of toxicants by using different endpoints.
4) In line with the 3Rs they would reduce the need for animals.
To 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,
Are you the right person for this PhD?
We are looking for an outstanding candidate with a high quality undergraduate or Masters degree (can be pending) in fields related to biochemistry, toxicology or cell biology and with the desire and drive to develop novel cell culture techniques to address the important question of adverse drug reactions in the liver and to apply these to safety testing.
This studentship starts in October 2017 and is funded by an MIBTP iCASE award which covers tuition fees and a competitive stipend for UK and EU nationals (where the EU nationals have lived in the UK for 3+ years). Deadline for applications January 8th 2017
When applying, include the name of the supervisor (Hodges), state the funding source (MIBTP iCASE) and the project title above.
For informal enquiries or for further information about the project please contact Nik Hodges (firstname.lastname@example.org)