| High-content screening of feeder-free human embryonic stem cells to identify pro-survival small molecules. | |
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MedLine Citation:
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PMID: 20854259 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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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:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: The Biochemical journal Volume: 432 ISSN: 1470-8728 ISO Abbreviation: Biochem. J. Publication Date: 2010 Nov |
Date Detail:
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Created Date: 2010-10-25 Completed Date: 2010-12-06 Revised Date: 2011-09-06 |
Medline Journal Info:
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Nlm Unique ID: 2984726R Medline TA: Biochem J Country: England |
Other Details:
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Languages: eng Pagination: 21-33 Citation Subset: IM |
Affiliation:
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University of Dundee, Scotland, UK. p.d.andrews@dundee.ac.uk |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amides
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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:
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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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