High content screening of feeder-free human embryonic stem cells to identify pro-survival small molecules

Andrews, P. et al. (2010) High content screening of feeder-free human embryonic stem cells to identify pro-survival small molecules. Biochemical Journal, 432(Pt 1), pp. 21-33. (doi:10.1042/BJ20101022)

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Publisher's URL: http://dx.doi.org/10.1042/BJ20101022

Abstract

The propensity of human embryonic stem cells to die upon enzymatic disaggregation or low-density plating is an obstacle to their isolation and routine use in drug discovery and basic research. Equally, the very low rate of establishment of implanted cells hinders cell therapy. Here we have developed a high-content assay for human embryonic stem cell survival and used this to screen a range of libraries of 'lead-like' small molecules and known bioactives. From this we identified 18 confirmed hits with 4 structural classes represented by multiple compounds: a series of 5-(acyl/alkyl-amino)indazoles, compounds with a 4-(acylamino)pyridine core, simple N6,N6-dialkyladenines and compounds with a 5-(acylamino)indolinone core. In vitro kinase profiling indicated that the ROCK/PRK2 protein kinases are of pivotal importance for cell survival and identified previously unreported compound classes with this important biological activity. Evaluation using an extensive panel of protein kinases showed that six of our hit compounds exhibited better selectivity for ROCK inhibition than the routinely used commercially available ROCK inhibitor, Y-27632. In this screen we also identified the K+-ATP channel opener, pinacidil, and show that it most likely promotes cell survival, by 'off-target' inhibition of ROCK/PRK2. We have identified novel pro-survival compounds of greater specificity, equivalent potency and reduced

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Houslay, Professor Miles and Mountford, Dr Joanne and Milligan, Professor Graeme
Authors: Andrews, P., Becroft, M., Aspegren, A., Gilmour, J., James, M., McRae, S., Kime, R., Allcock, R., Abraham, A., Jiang, Z., Strehl, R., Mountford, J.C., Milligan, G., Houslay, M.D., Adams, D., and Frearson, J.
College/School:College of Medical Veterinary and Life Sciences > Institute of Neuroscience and Psychology
Journal Name:Biochemical Journal
Publisher:Portland Press Ltd.
ISSN:0264-6021
ISSN (Online):1470-8728
Published Online:20 September 2010

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