Document Detail


High-content screening of feeder-free human embryonic stem cells to identify pro-survival small molecules.
MedLine Citation:
PMID:  20854259     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Paul D Andrews; Melissa Becroft; Anders Aspegren; Jane Gilmour; Martyn J James; Scott McRae; Robert Kime; Robert W Allcock; Achamma Abraham; Zhong Jiang; Raimund Strehl; Joanne C Mountford; Graeme Milligan; Miles D Houslay; David R Adams; Julie A Frearson
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Biochemical journal     Volume:  432     ISSN:  1470-8728     ISO Abbreviation:  Biochem. J.     Publication Date:  2010 Nov 
Date Detail:
Created Date:  2010-10-25     Completed Date:  2010-12-06     Revised Date:  2011-09-06    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  England    
Other Details:
Languages:  eng     Pagination:  21-33     Citation Subset:  IM    
Affiliation:
University of Dundee, Scotland, UK. p.d.andrews@dundee.ac.uk
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MeSH Terms
Descriptor/Qualifier:
Amides / pharmacology
Cell Culture Techniques
Cell Line
Cell Proliferation / drug effects*
Cell Survival / drug effects
Dose-Response Relationship, Drug
Drug Evaluation, Preclinical / methods
Embryonic Stem Cells / cytology,  drug effects*,  metabolism
Heterocyclic Compounds / chemistry,  pharmacology*
Humans
Indazoles / chemistry,  pharmacology
Molecular Structure
Protein Kinase C / antagonists & inhibitors,  metabolism
Protein Kinase Inhibitors / chemistry,  pharmacology*
Pyridines / chemistry,  pharmacology
Time Factors
rho-Associated Kinases / antagonists & inhibitors,  metabolism
Chemical
Reg. No./Substance:
0/Amides; 0/Heterocyclic Compounds; 0/Indazoles; 0/Protein Kinase Inhibitors; 0/Pyridines; 138381-45-0/Y 27632; EC 2.7.1.-/protein kinase N; EC 2.7.11.1/rho-Associated Kinases; EC 2.7.11.13/Protein Kinase C

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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