Development of a multi-cellular cardiac cell model to analyse drug-induced structural cardiotoxicity

Background:
Drug-induced cardiotoxicity is the leading cause of drug attrition in patients leading to drug withdrawal from the market or in pre-clinical development. Previous drug toxicity screens, developed in the 1990s, focussed on detecting adverse functional effects on cardiomyocyte electrophysiology. However, the advent of molecular targeted therapy with kinase inhibitors and therapeutic humanised antibodies has resulted in structural changes in multiple cardiac cells, which are not detected by current pre-clinical screens (Cross et al., 2015; Br. J. Pharmacol. 172:957-974.). There is a critical need for more advanced in vitro cell models of drug-induced cardiotoxicity to allow efficient and safe development of drugs by the pharmaceutical industry and access to more predictive advanced cell models across the SME and CRO communities.

Project Aims:
The aim is to build on existing cell biology strengths and develop an advanced human multi-cellular cell model of cardiac function to detect adverse structural changes in multiple cardiac cell types. We have already shown that a multi-cellular cardiac spheroid model composed of stem cell-derived cardiomyocytes, cardiac fibroblasts and cardiac microvascular endothelial cells can recapitulate in vivo cardiac physiology (Ravenscroft et al., 2016; Tox. Sci 152:99-112) and continues to be used utilised to analyse adverse functional changes in contractility in the CRO (Cyprotex) and Pharma (AstraZeneca) sectors. However, this current model does not show efficient vascularisation, which limits the ability to detect vascular structural changes, which are now becoming evident with kinase inhibitor drugs. This project will validate the suitability of the multi-cellular cardiac spheroid to detect adverse structural and functional changes in multiple cardiac cell types in response to a range of known and well annotated cardiotoxic drugs.

Scientific Objectives:
1. Develop current multicellular microtissue consisting of iPSC-CMs, human cardiac microvascular endothelial cells (HCMECs) and human cardiac fibroblasts (HCFs) to maximise development of all cell types. We will investigate specific growth factor additions and more physiological O2 levels to provide growth conditions which allow efficient vascularisation of microtissue. We will also investigate the potential for a 3D hydrogel scaffold to also enhance vascular physiology.
2. Investigate the potential to incorporate human cardiac pericytes into this model to improve recapitulation of human cardiac physiology. Improved cardiac physiology will also be monitored by mass spec analysis to compare with adult human cardiac tissue.
3. We will use the advanced fluorescence imaging technique light sheet microscopy to analyse changes in cardiac structure in 3D. This technique will allow us to simultaneously analyse changes in cardiomyocyte, fibroblast and endothelial cell morphology and directly visualise and quantitate cardiotoxic molecules.
4. Incorporation of cardiac microtissue into organ on a chip (OOAC) technology, to investigate model performance within a flow environment and in communication with other organoids.

Training and Support:
The successful candidate will join a vibrant research environment within the MRC Centre for Drug Safety Science (www.liv.ac.uk/drug-safety) in the Dept. of Molecular and Clinical Pharmacology, University of Liverpool. The student will gain extensive training in a number of techniques: primary human cell culture, cellular imaging and immunofluorescence, western blotting, gene expression and bioinformatic analysis. The project is also supported by funding from Medicine Discovery Catapult (MDC) based at Alderley Park Cheshire and the student will have the opportunity of an industrial placement within MDC.
Qualifications and Experience

Candidates should possess, or expect to obtain, a good degree (1st or 2:1) in a relevant discipline.
*Please note the English Language Requirement for EU Students is an IELTS score of 6.5 with no band score lower than 5.5.

Informal enquiries should be directed to Dr. Michael.Cross ([Email Address Removed]).
Applications to include full CV and the names of at least two academic referees should be sent to Dr. Michael Cross (E.mail: [Email Address Removed]) by closing date on 25th May 2018.