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Article: Genetic and pharmacological inhibition of PDK1 in cancer cells: Characterization of a selective allosteric kinase inhibitor

TitleGenetic and pharmacological inhibition of PDK1 in cancer cells: Characterization of a selective allosteric kinase inhibitor
Authors
Issue Date2011
PublisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/
Citation
Journal Of Biological Chemistry, 2011, v. 286 n. 8, p. 6433-6448 How to Cite?
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.
Persistent Identifierhttp://hdl.handle.net/10722/156005
ISSN
2014 Impact Factor: 4.573
2014 SCImago Journal Rankings: 2.734
PubMed Central ID
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorNagashima, Ken_US
dc.contributor.authorShumway, SDen_US
dc.contributor.authorSathyanarayanan, Sen_US
dc.contributor.authorChen, AHen_US
dc.contributor.authorDolinski, Ben_US
dc.contributor.authorXu, Yen_US
dc.contributor.authorKeilhack, Hen_US
dc.contributor.authorNguyen, Ten_US
dc.contributor.authorWiznerowicz, Men_US
dc.contributor.authorLi, Len_US
dc.contributor.authorLutterbach, BAen_US
dc.contributor.authorChi, Aen_US
dc.contributor.authorPaweletz, Cen_US
dc.contributor.authorAllison, Ten_US
dc.contributor.authorYan, Yen_US
dc.contributor.authorMunshi, SKen_US
dc.contributor.authorKlippel, Aen_US
dc.contributor.authorKraus, Men_US
dc.contributor.authorBobkova, EVen_US
dc.contributor.authorDeshmukh, Sen_US
dc.contributor.authorXu, Zen_US
dc.contributor.authorMueller, Uen_US
dc.contributor.authorSzewczak, AAen_US
dc.contributor.authorPan, BSen_US
dc.contributor.authorRichon, Ven_US
dc.contributor.authorPollock, Ren_US
dc.contributor.authorBlumeJensen, Pen_US
dc.contributor.authorNorthrup, Aen_US
dc.contributor.authorAndersen, JNen_US
dc.date.accessioned2012-08-08T08:39:27Z-
dc.date.available2012-08-08T08:39:27Z-
dc.date.issued2011en_US
dc.identifier.citationJournal Of Biological Chemistry, 2011, v. 286 n. 8, p. 6433-6448en_US
dc.identifier.issn0021-9258en_US
dc.identifier.urihttp://hdl.handle.net/10722/156005-
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.en_US
dc.languageengen_US
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/en_US
dc.relation.ispartofJournal of Biological Chemistryen_US
dc.subject.meshAllosteric Regulation - Drug Effects - Geneticsen_US
dc.subject.meshAnimalsen_US
dc.subject.meshCatalytic Domain - Geneticsen_US
dc.subject.meshCell Line, Tumoren_US
dc.subject.meshCell Proliferation - Drug Effectsen_US
dc.subject.meshCrystallography, X-Rayen_US
dc.subject.meshDogsen_US
dc.subject.meshDrug Screening Assays, Antitumor - Methodsen_US
dc.subject.meshHumansen_US
dc.subject.meshNeoplasm Proteins - Antagonists & Inhibitors - Genetics - Metabolismen_US
dc.subject.meshNeoplasms - Drug Therapy - Enzymology - Geneticsen_US
dc.subject.meshPhosphorylation - Drug Effects - Geneticsen_US
dc.subject.meshProtein Kinase Inhibitors - Chemistry - Pharmacologyen_US
dc.subject.meshProtein-Serine-Threonine Kinases - Antagonists & Inhibitors - Genetics - Metabolismen_US
dc.titleGenetic and pharmacological inhibition of PDK1 in cancer cells: Characterization of a selective allosteric kinase inhibitoren_US
dc.typeArticleen_US
dc.identifier.emailChen, AH:hrllchy@hkucc.hku.hken_US
dc.identifier.authorityChen, AH=rp01240en_US
dc.description.naturelink_to_OA_fulltexten_US
dc.identifier.doi10.1074/jbc.M110.156463en_US
dc.identifier.pmid21118801en_US
dc.identifier.pmcidPMC3057779-
dc.identifier.scopuseid_2-s2.0-79953181999en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-79953181999&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume286en_US
dc.identifier.issue8en_US
dc.identifier.spage6433en_US
dc.identifier.epage6448en_US
dc.identifier.isiWOS:000287476400054-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridNagashima, K=35741383200en_US
dc.identifier.scopusauthoridShumway, SD=7004980416en_US
dc.identifier.scopusauthoridSathyanarayanan, S=6506689732en_US
dc.identifier.scopusauthoridChen, AH=7403392103en_US
dc.identifier.scopusauthoridDolinski, B=7801344570en_US
dc.identifier.scopusauthoridXu, Y=45761619500en_US
dc.identifier.scopusauthoridKeilhack, H=6603161345en_US
dc.identifier.scopusauthoridNguyen, T=35564962000en_US
dc.identifier.scopusauthoridWiznerowicz, M=6602377756en_US
dc.identifier.scopusauthoridLi, L=45761266600en_US
dc.identifier.scopusauthoridLutterbach, BA=36941446500en_US
dc.identifier.scopusauthoridChi, A=8695748800en_US
dc.identifier.scopusauthoridPaweletz, C=26638385600en_US
dc.identifier.scopusauthoridAllison, T=15022029300en_US
dc.identifier.scopusauthoridYan, Y=7404585684en_US
dc.identifier.scopusauthoridMunshi, SK=7005735959en_US
dc.identifier.scopusauthoridKlippel, A=7004112507en_US
dc.identifier.scopusauthoridKraus, M=7202455637en_US
dc.identifier.scopusauthoridBobkova, EV=7004542598en_US
dc.identifier.scopusauthoridDeshmukh, S=7102221634en_US
dc.identifier.scopusauthoridXu, Z=35742577800en_US
dc.identifier.scopusauthoridMueller, U=12773511000en_US
dc.identifier.scopusauthoridSzewczak, AA=36837779200en_US
dc.identifier.scopusauthoridPan, BS=45761375800en_US
dc.identifier.scopusauthoridRichon, V=35312046200en_US
dc.identifier.scopusauthoridPollock, R=7102871810en_US
dc.identifier.scopusauthoridBlumeJensen, P=6602824234en_US
dc.identifier.scopusauthoridNorthrup, A=6506550319en_US
dc.identifier.scopusauthoridAndersen, JN=35512446300en_US

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