File Download
 
Links for fulltext
(May Require Subscription)
 
Supplementary

Article: Genetic and pharmacological inhibition of PDK1 in cancer cells: Characterization of a selective allosteric kinase inhibitor
  • Basic View
  • Metadata View
  • XML View
TitleGenetic and pharmacological inhibition of PDK1 in cancer cells: Characterization of a selective allosteric kinase inhibitor
 
AuthorsNagashima, K1
Shumway, SD1
Sathyanarayanan, S1
Chen, AH1
Dolinski, B1
Xu, Y1
Keilhack, H1
Nguyen, T1
Wiznerowicz, M1
Li, L1
Lutterbach, BA1
Chi, A1
Paweletz, C1
Allison, T2
Yan, Y2
Munshi, SK2
Klippel, A1
Kraus, M1
Bobkova, EV1
Deshmukh, S1
Xu, Z1
Mueller, U1
Szewczak, AA1
Pan, BS1
Richon, V1
Pollock, R1
BlumeJensen, P1
Northrup, A1
Andersen, JN1
 
Issue Date2011
 
PublisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/
 
CitationJournal Of Biological Chemistry, 2011, v. 286 n. 8, p. 6433-6448 [How to Cite?]
DOI: http://dx.doi.org/10.1074/jbc.M110.156463
 
AbstractPhosphoinositide-dependent kinase 1 (PDK1) is a critical activator of multiple prosurvival and oncogenic protein kinases and has garnered considerable interest as an oncology drug target. Despite progress characterizing PDK1 as a therapeutic target, pharmacological support is lacking due to the prevalence of nonspecific inhibitors. Here, we benchmark literature and newly developed inhibitors and conduct parallel genetic and pharmacological queries into PDK1 function in cancer cells. Through kinase selectivity profiling and x-ray crystallographic studies, we identify an exquisitely selective PDK1 inhibitor (compound 7) that uniquely binds to the inactive kinase conformation (DFG-out). In contrast to compounds 1-5, which are classical ATP-competitive kinase inhibitors (DFG-in), compound 7 specifically inhibits cellular PDK1 T-loop phosphorylation (Ser-241), supporting its unique binding mode. Interfering with PDK1 activity has minimal antiproliferative effect on cells growing as plastic-attached monolayer cultures (i.e. standard tissue culture conditions) despite reduced phosphorylation of AKT, RSK, and S6RP. However, selective PDK1 inhibition impairs anchorage-independent growth, invasion, and cancer cell migration. Compound 7 inhibits colony formation in a subset of cancer cell lines (four of 10) and primary xenograft tumor lines (nine of 57). RNAi-mediated knockdown corroborates the PDK1 dependence in cell lines and identifies candidate biomarkers of drug response. In summary, our profiling studies define a uniquely selective and cell-potent PDK1 inhibitor, and the convergence of genetic and pharmacological phenotypes supports a role of PDK1 in tumorigenesis in the context of three-dimensional in vitro culture systems. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.
 
ISSN0021-9258
2013 Impact Factor: 4.600
 
DOIhttp://dx.doi.org/10.1074/jbc.M110.156463
 
ISI Accession Number IDWOS:000287476400054
 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorNagashima, K
 
dc.contributor.authorShumway, SD
 
dc.contributor.authorSathyanarayanan, S
 
dc.contributor.authorChen, AH
 
dc.contributor.authorDolinski, B
 
dc.contributor.authorXu, Y
 
dc.contributor.authorKeilhack, H
 
dc.contributor.authorNguyen, T
 
dc.contributor.authorWiznerowicz, M
 
dc.contributor.authorLi, L
 
dc.contributor.authorLutterbach, BA
 
dc.contributor.authorChi, A
 
dc.contributor.authorPaweletz, C
 
dc.contributor.authorAllison, T
 
dc.contributor.authorYan, Y
 
dc.contributor.authorMunshi, SK
 
dc.contributor.authorKlippel, A
 
dc.contributor.authorKraus, M
 
dc.contributor.authorBobkova, EV
 
dc.contributor.authorDeshmukh, S
 
dc.contributor.authorXu, Z
 
dc.contributor.authorMueller, U
 
dc.contributor.authorSzewczak, AA
 
dc.contributor.authorPan, BS
 
dc.contributor.authorRichon, V
 
dc.contributor.authorPollock, R
 
dc.contributor.authorBlumeJensen, P
 
dc.contributor.authorNorthrup, A
 
dc.contributor.authorAndersen, JN
 
dc.date.accessioned2012-08-08T08:39:27Z
 
dc.date.available2012-08-08T08:39:27Z
 
dc.date.issued2011
 
dc.description.abstractPhosphoinositide-dependent kinase 1 (PDK1) is a critical activator of multiple prosurvival and oncogenic protein kinases and has garnered considerable interest as an oncology drug target. Despite progress characterizing PDK1 as a therapeutic target, pharmacological support is lacking due to the prevalence of nonspecific inhibitors. Here, we benchmark literature and newly developed inhibitors and conduct parallel genetic and pharmacological queries into PDK1 function in cancer cells. Through kinase selectivity profiling and x-ray crystallographic studies, we identify an exquisitely selective PDK1 inhibitor (compound 7) that uniquely binds to the inactive kinase conformation (DFG-out). In contrast to compounds 1-5, which are classical ATP-competitive kinase inhibitors (DFG-in), compound 7 specifically inhibits cellular PDK1 T-loop phosphorylation (Ser-241), supporting its unique binding mode. Interfering with PDK1 activity has minimal antiproliferative effect on cells growing as plastic-attached monolayer cultures (i.e. standard tissue culture conditions) despite reduced phosphorylation of AKT, RSK, and S6RP. However, selective PDK1 inhibition impairs anchorage-independent growth, invasion, and cancer cell migration. Compound 7 inhibits colony formation in a subset of cancer cell lines (four of 10) and primary xenograft tumor lines (nine of 57). RNAi-mediated knockdown corroborates the PDK1 dependence in cell lines and identifies candidate biomarkers of drug response. In summary, our profiling studies define a uniquely selective and cell-potent PDK1 inhibitor, and the convergence of genetic and pharmacological phenotypes supports a role of PDK1 in tumorigenesis in the context of three-dimensional in vitro culture systems. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.
 
dc.description.natureLink_to_subscribed_fulltext
 
dc.identifier.citationJournal Of Biological Chemistry, 2011, v. 286 n. 8, p. 6433-6448 [How to Cite?]
DOI: http://dx.doi.org/10.1074/jbc.M110.156463
 
dc.identifier.doihttp://dx.doi.org/10.1074/jbc.M110.156463
 
dc.identifier.epage6448
 
dc.identifier.isiWOS:000287476400054
 
dc.identifier.issn0021-9258
2013 Impact Factor: 4.600
 
dc.identifier.issue8
 
dc.identifier.pmid21118801
 
dc.identifier.scopuseid_2-s2.0-79953181999
 
dc.identifier.spage6433
 
dc.identifier.urihttp://hdl.handle.net/10722/156005
 
dc.identifier.volume286
 
dc.languageeng
 
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/
 
dc.publisher.placeUnited States
 
dc.relation.ispartofJournal of Biological Chemistry
 
dc.relation.referencesReferences in Scopus
 
dc.subject.meshAllosteric Regulation - Drug Effects - Genetics
 
dc.subject.meshAnimals
 
dc.subject.meshCatalytic Domain - Genetics
 
dc.subject.meshCell Line, Tumor
 
dc.subject.meshCell Proliferation - Drug Effects
 
dc.subject.meshCrystallography, X-Ray
 
dc.subject.meshDogs
 
dc.subject.meshDrug Screening Assays, Antitumor - Methods
 
dc.subject.meshHumans
 
dc.subject.meshNeoplasm Proteins - Antagonists & Inhibitors - Genetics - Metabolism
 
dc.subject.meshNeoplasms - Drug Therapy - Enzymology - Genetics
 
dc.subject.meshPhosphorylation - Drug Effects - Genetics
 
dc.subject.meshProtein Kinase Inhibitors - Chemistry - Pharmacology
 
dc.subject.meshProtein-Serine-Threonine Kinases - Antagonists & Inhibitors - Genetics - Metabolism
 
dc.titleGenetic and pharmacological inhibition of PDK1 in cancer cells: Characterization of a selective allosteric kinase inhibitor
 
dc.typeArticle
 
<?xml encoding="utf-8" version="1.0"?>
<item><contributor.author>Nagashima, K</contributor.author>
<contributor.author>Shumway, SD</contributor.author>
<contributor.author>Sathyanarayanan, S</contributor.author>
<contributor.author>Chen, AH</contributor.author>
<contributor.author>Dolinski, B</contributor.author>
<contributor.author>Xu, Y</contributor.author>
<contributor.author>Keilhack, H</contributor.author>
<contributor.author>Nguyen, T</contributor.author>
<contributor.author>Wiznerowicz, M</contributor.author>
<contributor.author>Li, L</contributor.author>
<contributor.author>Lutterbach, BA</contributor.author>
<contributor.author>Chi, A</contributor.author>
<contributor.author>Paweletz, C</contributor.author>
<contributor.author>Allison, T</contributor.author>
<contributor.author>Yan, Y</contributor.author>
<contributor.author>Munshi, SK</contributor.author>
<contributor.author>Klippel, A</contributor.author>
<contributor.author>Kraus, M</contributor.author>
<contributor.author>Bobkova, EV</contributor.author>
<contributor.author>Deshmukh, S</contributor.author>
<contributor.author>Xu, Z</contributor.author>
<contributor.author>Mueller, U</contributor.author>
<contributor.author>Szewczak, AA</contributor.author>
<contributor.author>Pan, BS</contributor.author>
<contributor.author>Richon, V</contributor.author>
<contributor.author>Pollock, R</contributor.author>
<contributor.author>BlumeJensen, P</contributor.author>
<contributor.author>Northrup, A</contributor.author>
<contributor.author>Andersen, JN</contributor.author>
<date.accessioned>2012-08-08T08:39:27Z</date.accessioned>
<date.available>2012-08-08T08:39:27Z</date.available>
<date.issued>2011</date.issued>
<identifier.citation>Journal Of Biological Chemistry, 2011, v. 286 n. 8, p. 6433-6448</identifier.citation>
<identifier.issn>0021-9258</identifier.issn>
<identifier.uri>http://hdl.handle.net/10722/156005</identifier.uri>
<description.abstract>Phosphoinositide-dependent kinase 1 (PDK1) is a critical activator of multiple prosurvival and oncogenic protein kinases and has garnered considerable interest as an oncology drug target. Despite progress characterizing PDK1 as a therapeutic target, pharmacological support is lacking due to the prevalence of nonspecific inhibitors. Here, we benchmark literature and newly developed inhibitors and conduct parallel genetic and pharmacological queries into PDK1 function in cancer cells. Through kinase selectivity profiling and x-ray crystallographic studies, we identify an exquisitely selective PDK1 inhibitor (compound 7) that uniquely binds to the inactive kinase conformation (DFG-out). In contrast to compounds 1-5, which are classical ATP-competitive kinase inhibitors (DFG-in), compound 7 specifically inhibits cellular PDK1 T-loop phosphorylation (Ser-241), supporting its unique binding mode. Interfering with PDK1 activity has minimal antiproliferative effect on cells growing as plastic-attached monolayer cultures (i.e. standard tissue culture conditions) despite reduced phosphorylation of AKT, RSK, and S6RP. However, selective PDK1 inhibition impairs anchorage-independent growth, invasion, and cancer cell migration. Compound 7 inhibits colony formation in a subset of cancer cell lines (four of 10) and primary xenograft tumor lines (nine of 57). RNAi-mediated knockdown corroborates the PDK1 dependence in cell lines and identifies candidate biomarkers of drug response. In summary, our profiling studies define a uniquely selective and cell-potent PDK1 inhibitor, and the convergence of genetic and pharmacological phenotypes supports a role of PDK1 in tumorigenesis in the context of three-dimensional in vitro culture systems. &#169; 2011 by The American Society for Biochemistry and Molecular Biology, Inc.</description.abstract>
<language>eng</language>
<publisher>American Society for Biochemistry and Molecular Biology, Inc. The Journal&apos;s web site is located at http://www.jbc.org/</publisher>
<relation.ispartof>Journal of Biological Chemistry</relation.ispartof>
<subject.mesh>Allosteric Regulation - Drug Effects - Genetics</subject.mesh>
<subject.mesh>Animals</subject.mesh>
<subject.mesh>Catalytic Domain - Genetics</subject.mesh>
<subject.mesh>Cell Line, Tumor</subject.mesh>
<subject.mesh>Cell Proliferation - Drug Effects</subject.mesh>
<subject.mesh>Crystallography, X-Ray</subject.mesh>
<subject.mesh>Dogs</subject.mesh>
<subject.mesh>Drug Screening Assays, Antitumor - Methods</subject.mesh>
<subject.mesh>Humans</subject.mesh>
<subject.mesh>Neoplasm Proteins - Antagonists &amp; Inhibitors - Genetics - Metabolism</subject.mesh>
<subject.mesh>Neoplasms - Drug Therapy - Enzymology - Genetics</subject.mesh>
<subject.mesh>Phosphorylation - Drug Effects - Genetics</subject.mesh>
<subject.mesh>Protein Kinase Inhibitors - Chemistry - Pharmacology</subject.mesh>
<subject.mesh>Protein-Serine-Threonine Kinases - Antagonists &amp; Inhibitors - Genetics - Metabolism</subject.mesh>
<title>Genetic and pharmacological inhibition of PDK1 in cancer cells: Characterization of a selective allosteric kinase inhibitor</title>
<type>Article</type>
<description.nature>Link_to_subscribed_fulltext</description.nature>
<identifier.doi>10.1074/jbc.M110.156463</identifier.doi>
<identifier.pmid>21118801</identifier.pmid>
<identifier.scopus>eid_2-s2.0-79953181999</identifier.scopus>
<relation.references>http://www.scopus.com/mlt/select.url?eid=2-s2.0-79953181999&amp;selection=ref&amp;src=s&amp;origin=recordpage</relation.references>
<identifier.volume>286</identifier.volume>
<identifier.issue>8</identifier.issue>
<identifier.spage>6433</identifier.spage>
<identifier.epage>6448</identifier.epage>
<identifier.isi>WOS:000287476400054</identifier.isi>
<publisher.place>United States</publisher.place>
</item>
Author Affiliations
  1. Merck Research Laboratories
  2. Merck &amp; Co.