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

Andrews, Paul D., Becroft, Melissa, Aspegren, Anders, Gilmour, Jane, James, Martyn J., McRae, Scott, Kime, Robert, Allcock, Robert W., Abraham, Achamma, Jiang, Zhong, Strehl, Raimund, Mountford, Joanne C., Milligan, Graeme, Houslay, Miles D., Adams, David R. and Frearson, Julie A. (2010) High-content screening of feeder-free human embryonic stem cells to identify pro-survival small molecules. Biochemical Journal, 432 1: 21-33. doi:10.1042/BJ20101022


Author Andrews, Paul D.
Becroft, Melissa
Aspegren, Anders
Gilmour, Jane
James, Martyn J.
McRae, Scott
Kime, Robert
Allcock, Robert W.
Abraham, Achamma
Jiang, Zhong
Strehl, Raimund
Mountford, Joanne C.
Milligan, Graeme
Houslay, Miles D.
Adams, David R.
Frearson, Julie A.
Title High-content screening of feeder-free human embryonic stem cells to identify pro-survival small molecules
Journal name Biochemical Journal   Check publisher's open access policy
ISSN 0264-6021
1470-8728
Publication date 2010-01-01
Sub-type Article (original research)
DOI 10.1042/BJ20101022
Open Access Status Not Open Access
Volume 432
Issue 1
Start page 21
End page 33
Total pages 13
Place of publication London, United Kingdom
Publisher Portland Press
Language eng
Formatted 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. In the present study 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 four structural classes being 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 (Rho-associated kinase)/PRK2 (protein kinase C-related kinase 2) protein kinases are of pivotal importance for cell survival and identified previously unreported compound classes that inhibited this important biological activity.An 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 probably promotes cell survival, by 'off-target' inhibition of ROCK/PRK2. We have therefore identified novel pro-survival compounds of greater specificity, equivalent potency and reduced toxicity relative to the routinely employed ROCK inhibitor Y-27632.
Keyword Drug discovery
High-content screening
Human embryonic stem cell
Pinacidil
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Institute for Molecular Bioscience - Publications
 
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