Creation of bio-artificial kidney with renal cells (primary, immortalised or stem) as a model of renal transport

Department Name: Centre for Biological Engineering at Loughborough University and DMPK AstraZeneca
Supervisors: Dr Karen Coopman and Dr Elnaz Gozalpour

Development of an in vitro human renal model is a key requirement for early stage drug development to predict renal clearance in human and potentially develop structure activity relationships. The aim of this project is to generate such a model which can also enable the determination in vitro to in vivo extrapolation (IVIVE) for renal clearance and drug-drug interactions (DDIs) via renal transporters. Key factors for successful development of an in vitro renal model are expression and function of renal transporters along with formation of tight cellular monolayer where both apical and basolateral compartments that can be accessed and sampled separately.

A bioreactor system utilising polysulphone hollow fibres has previously been developed at Loughborough University. Tubular orientation, application of flow to mimic the in vivo environment of the proximal tubule and the structural composition of bioreactor system have been shown to improve proximal tubule cell growth and viability, however, the effect of this platform on expression and function renal uptake and efflux transporters remains to be studied.

In this project, human kidney cells (HK2) are therefore going to be used to determine the optimum conditions in terms of cell culture medium composition, cell density, flow rate, cell culture surface on the polysulphone fibres and extrusion method to produce consistent hollow fibres for renal cells growth on the bioreactor system. Subsequently, human renal proximal tubule cells including primary and immortalized cells will be tested in the bioreactor to assess the uptake and efflux transporters expression, function and formation of tight cellular monolayer. Once cell types with optimum growth, viability, and renal transporter expression in the bioreactor system are selected they will be ultilised as an in vitro human renal model to determine an IVIVE for renal clearance, prediction of renal secretion and drug-drug interactions via renal transporters. The data produced by this model may also be included in physiologically based pharmacokinetic modelling packages to evaluate in vivo DDIs prediction in human.

Different cell culture techniques, protein expression analysis via western blotting and confocal microscopy, proteomics, transporters functional assay such as cellular uptake and efflux assays, transepithelial flux assay, and bioanalysis methods using fluorescent and radiochemical compounds and mass spectrometry may be used in this project.

Applicants must hold, or expect to receive, a first or upper second class honours degree (or equivalent) in chemistry, biochemistry, cell and molecular biology, pharmacology or biochemical/medical engineering with knowledge of organic chemistry. Desirable knowledge includes spectroscopy, cell culture and cell characterisation assays and candidates must possess effective communication and team working skills. The successful candidate will work primarily in the Centre for Biological Engineering at Loughborough University with secondments to AstraZeneca as the work requires. He/she will be co-supervised by Dr Karen Coopman and Dr. Elnaz Gozalpour.

The PhD position is a CASE studentship based at Centre for Biological Engineering, Loughborough University for 4 years . The project is a scientific collaboration between a team at Loughborough and a team at AstraZeneca and the successful applicant will also spend time at the AstraZeneca site in Cambridge as necessary for progression of the project.