Human pluripotent stem cell cardiomyocytes and hepatocytes with engineered genotypes for drug safety evaluation

Safety assessment of each new drug developed requires up to 1800 animals, which typically comprise non-rodent species (monkeys, dogs) and rodents (rats and mice). New EU regulations on toxicity testing (termed 'REACH') will use up to 54 million animals over the next 10 years to evaluate 30,000 compounds. Even with this level of animal use, preclinical assays are cited as only 71% predictive of whether a drug will be toxic in humans. Poor predictability is due to species differences and the inability to develop test platforms that mirror diverse human genotypes, particularly those associated with the high drug susceptibility seen in cardio- and hepato-toxicity. Adverse drug reactions account for 100,000 deaths per year in the US alone. To facilitate reduction and replacement of animal use, and to increase predictability to human toxicity, this proposal brings together a new consortium with skills in genome engineering (Skarnes, Rosen; Sanger Centre), human pluripotent stem cell biology and robotic automation (hPSC; Denning, Young; Nottingham) and toxicity testing (Goldring, Park; Liverpool). The aim is to engineer different patient-relevant mutations associated with drug susceptibility or resistance into the protein coding regions of otherwise genetically healthy hPSCs. Differentiating these cells will produce cardiomyocytes and hepatocytes that carry specific mutations within a common genetic background, allowing unbiased evaluation of how genotype-drug interaction affects cell structure, function and viability. Co-culturing cardiomyocytes with hepatocytes of different genotypes will allow the impact of altered hepatocyte function on cardiomyocytes to be assessed. Comparing these results with data from the literature and industrial partners will allow the predictive value of this humanised in vitro model to be determined. A 0.1% reduction in animal-based in vitro, ex vivo and in vivo tests has the potential to save up to 54,000 animals in Europe alone.

Hall C et al. (2021). Complex Relationship Between Cardiac Fibroblasts and Cardiomyocytes in Health and Disease. Journal of the American Heart Association 10: e019338. doi: 10.1161/JAHA.120.019338

Bhagwan JR et al. (2020). Isogenic models of hypertrophic cardiomyopathy unveil differential phenotypes and mechanism-driven therapeutics. Journal of Molecular and Cellular Cardiology 145: 43–53. doi: 10.1016/j.yjmcc.2020.06.003

Kondrashov et al. (2020). CRISPR/Cas9-mediated generation and analysis of N terminus polymorphic models of β2AR in isogenic hPSC-derived cardiomyocytes. Molecular Therapy Methods & Clinical Development 20:39-53. doi: 10.1016/j.omtm.2020.10.019

Saleem U et al. (2020). Force and Calcium Transients Analysis in Human Engineered Heart Tissues Reveals Positive Force-Frequency Relation at Physiological Frequency. Stem Cell Reports 14(2):312-24. doi: 10.1016/j.stemcr.2019.12.011

Abakir A et al. (2019). N6-methyladenosine regulates the stability of RNA:DNA hybrids in human cells. Nature Genetics 52:48-55. doi: 10.1038/s41588-019-0549-x

Alvarez-Paino M et al. (2019). Polymer Microparticles with Defined Surface Chemistry and Topography Mediate the Formation of Stem Cell Aggregates and Cardiomyocyte Function. ACS Appl. Mater. Interfaces 11(38):34560–34574. doi: 10.1021/acsami.9b04769

de Korte T et al. (2019). Unlocking Personalized Biomedicine and Drug Discovery with Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes: Fit for Purpose or Forever Elusive? Annual Review of Pharmacology and Toxicology 60:529-551. doi: 10.1146/annurev-pharmtox-010919-023309

Howe CL et al. (2019). Surface plasmon resonance imaging of excitable cells. Journal of Physics D: Applied Physics 52(10):104001. doi: 10.1088/1361-6463/aaf849

Mosqueira D et al. (2019). High-Throughput Phenotyping Toolkit for Characterizing Cellular Models of Hypertrophic Cardiomyopathy In VitroMethods and Protocols 2(4):83. doi: 10.3390/mps2040083

Mosqueira D et al. (2019). Modeling Hypertrophic Cardiomyopathy: Mechanistic Insights and Pharmacological Intervention. Trends in Molecular Medicine 25(9):775-790. doi: 10.1016/j.molmed.2019.06.005

Vaithilingam J et al. (2019). Multifunctional Bioinstructive 3D Architectures to Modulate Cellular Behavior. Advanced Functional Materials 29(38):1902016. doi: 10.1002/adfm.201902016

van Meer BJ et al. (2019). Simultaneous measurement of excitation-contraction coupling parameters identifies mechanisms underlying contractile responses of hiPSC-derived cardiomyocytes. Nature Communications. 10:4325. doi: 10.1038/s41467-019-12354-8

Zhou X et al. (2019). Investigating the Complex Arrhythmic Phenotype Caused by the Gain-of-Function Mutation KCNQ1-G229D. Frontiers in Physiology 10:259. doi: 10.3389/fphys.2019.00259

Kondrashov A et al. (2018). Simplified Footprint-Free Cas9/CRISPR Editing of Cardiac-Associated Genes in Human Pluripotent Stem Cells. Stem Cells and Development 27(6):391-404. doi: 10.1089/scd.2017.0268

Mosqueira D et al. (2018). CRISPR/Cas9 editing in human pluripotent stem cell-cardiomyocytes highlights arrhythmias, hypocontractility, and energy depletion as potential therapeutic targets for hypertrophic cardiomyopathy. European Heart Journal 39(43):3879-91. doi: 10.1093/eurheartj/ehy249

Smith JGW et al. (2018). Isogenic Pairs of hiPSC-CMs with Hypertrophic Cardiomyopathy/LVNC-Associated ACTC1 E99K Mutation Unveil Differential Functional Deficits. Stem Cell Reports 11(5):1226-1243. doi: 10.1016/j.stemcr.2018.10.006

Duncan G et al. (2017). Drug-Mediated Shortening of Action Potentials in LQTS2 Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes. Stem Cells and Development 26(23):1695-1705. doi: 10.1089/scd.2017.0172

Goldring C et al. (2017). Stem cell-derived models to improve mechanistic understanding and prediction of human drug-induced liver injury. Hepatology 65(2):71-21. doi: 10.1002/hep.28886

Denning C et al. (2016). Cardiomyocytes from human pluripotent stem cells: From laboratory curiosity to industrial biomedical platform. Biochim Biophys Acta. 1863(7 Pt B):1728-48. doi: 10.1016/j.bbamcr.2015.10.014

Dixon JE et al. (2016). Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides. PNAS 113(3):E291-9. doi: 10.1073/pnas.1518634113

Hammad M et al. (2016). Identification of polymer surface adsorbed proteins implicated in pluripotent human embryonic stem cell expansion. Biomaterials Science 4(9):1381-91. doi: 10.1039/c6bm00214e

Kalra S et al. (2016). Can Human Pluripotent Stem Cell-Derived Cardiomyocytes Advance Understanding of Muscular Dystrophies? Journal of Neuromuscular Diseases 3(3):309-32. doi: 10.3233/JND-150133

Patel A et al. (2016). High throughput screening for discovery of materials that control stem cell fate. Current Opinion in Solid State and Materials Science 20(4) doi: 10.1016/j.cossms.2016.02.002

Rajamohan D et al. (2016). Automated Electrophysiological and Pharmacological Evaluation of Human Pluripotent Stem Cell-Derived Cardiomyocytes. Stem Cells and Development 25(6):439-52. doi: 10.1089/scd.2015.0253

Celiz AD et al. (2015). Discovery of a Novel Polymer for Human Pluripotent Stem Cell Expansion and Multilineage Differentiation. Advanced Materials 27(27):4006-12. doi: 10.1002/adma.201501351

Lin B et al. (2015). Modeling and study of the mechanism of dilated cardiomyopathy using induced pluripotent stem cells derived from individuals with Duchenne muscular dystrophy. Dis Model Mech. 8(5):457-66. doi: 10.1242/dmm.019505 

Patel AK et al. (2015). A defined synthetic substrate for serum-free culture of human stem cell derived cardiomyocytes with improved functional maturity identified using combinatorial materials microarrays. Biomaterials 61:257-65. doi:10.1016/j.biomaterials.2015.05.019

Ribeiro MC et al. (2015). Functional maturation of human pluripotent stem cell derived cardiomyocytes in vitro--correlation between contraction force and electrophysiology. 51(138-150). doi: 10.1016/j.biomaterials.2015.01.067

Smith JG et al. (2015). Scaling human pluripotent stem cell expansion and differentiation: are cell factories becoming a reality? Regen Med. 10(8):925-30. doi: 10.2217/rme.15.65

Celiz AD et al. (2014). Chemically diverse polymer microarrays and high throughput surface characterisation: a method for discovery of materials for stem cell culture. Biomaterials Science 2(11):1604-11. doi: 10.1039/c4bm00054d

Celiz AD et al. (2014). Materials for stem cell factories of the future. Nature Materials 13(6):570-9. doi: 10.1038/nmat3972

Dixon JE et al. (2014). Combined hydrogels that switch human pluripotent stem cells from self-renewal to differentiation. PNAS 111(15):5580-5. doi: 10.1073/pnas.1319685111

Földes G et al. (2014). Aberrant α-adrenergic hypertrophic response in cardiomyocytes from human induced pluripotent cells. Stem Cell Reports. 3(5):905-14. doi:10.1016/j.stemcr.2014.09.002

Matsa E et al. (2014). Allele-specific RNA interference rescues the long-QT syndrome phenotype in human-induced pluripotency stem cell cardiomyocytes. Eur Heart J. 35(16):1078-87. doi: 10.1093/eurheartj/eht067

Back to top
Project grant



Principal investigator

Professor Chris Denning


University of Nottingham


Professor Lorraine Young
Dr Chris Goldring
Professor Kevin Park
Dr William Skarnes

Grant reference number


Award date

Apr 2013 - Apr 2016

Grant amount