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.

Denning C, Borgdorff V, Crutchley J, Firth KS, George V, Kalra S, Kondrashov A, Hoang MD, Mosqueira D, Patel A, Prodanov L, Rajamohan D, Skarnes WC, Smith JG, Young LE (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

Smith JG, Celiz AD, Patel AK, Short RD, Alexander MR, Denning C (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

Patel AK, Celiz AD, Rajamohan D, Anderson DG, Langer R, Davies MC, Alexander MR, Denning C (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

Lin B, Li Y, Han L, Kaplan AD, Ao Y, Kalra S, Bett GC, Rasmusson RL, Denning C, Yang L (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 

Matsa E, Dixon JE, Medway C, Georgiou O, Patel MJ, Morgan K, Kemp PJ, Staniforth A, Mellor I, Denning C (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

Földes G, Matsa E, Kriston-Vizi J, Leja T, Amisten S, Kolker L, Kodagoda T, Dolatshad NF, Mioulane M, Vauchez K, Arányi T, Ketteler R, Schneider MD, Denning C, Harding SE (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

Dick E, Kalra S, Anderson D, George V, Ritson M, Laval S, Barresi R, Aartsma-Rus A, Lochmuller H, Denning C (2013). Exon skipping and gene transfer restore dystrophin expression in hiPSC-cardiomyocytes harbouring DMD mutations. Stem Cells Dev. doi: 10.1089/2013.0135

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Apr 2013 - Apr 2016

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