title: GSK-3β Governs Inflammation-Induced NFATc2 Signaling Hubs to Promote Pancreatic Cancer Progression.
abstract: We aimed to investigate the mechanistic, functional, and therapeutic role of glycogen synthase kinase 3β (GSK-3β) in the regulation and activation of the proinflammatory oncogenic transcription factor nuclear factor of activated T cells (NFATc2) in pancreatic cancer. IHC, qPCR, immunoblotting, immunofluorescence microscopy, and proliferation assays were used to analyze mouse and human tissues and cell lines. Protein-protein interactions and promoter regulation were analyzed by coimmunoprecipitation, DNA pulldown, reporter, and ChIP assays. Preclinical assays were performed using a variety of pancreatic cancer cells lines, xenografts, and a genetically engineered mouse model (GEMM). GSK-3β-dependent SP2 phosphorylation mediates NFATc2 protein stability in the nucleus of pancreatic cancer cells stimulating pancreatic cancer growth. In addition to protein stabilization, GSK-3β also maintains NFATc2 activation through a distinct mechanism involving stabilization of NFATc2-STAT3 complexes independent of SP2 phosphorylation. For NFATc2-STAT3 complex formation, GSK-3β-mediated phosphorylation of STAT3 at Y705 is required to stimulate euchromatin formation of NFAT target promoters, such as cyclin-dependent kinase-6, which promotes tumor growth. Finally, preclinical experiments suggest that targeting the NFATc2-STAT3-GSK-3β module inhibits proliferation and tumor growth and interferes with inflammation-induced pancreatic cancer progression in Kras(G12D) mice. In conclusion, we describe a novel mechanism by which GSK-3β fine-tunes NFATc2 and STAT3 transcriptional networks to integrate upstream signaling events that govern pancreatic cancer progression and growth. Furthermore, the therapeutic potential of GSK-3β is demonstrated for the first time in a relevant Kras and inflammation-induced GEMM for pancreatic cancer.
authors.affiliation: Department of Gastroenterology, Endocrinology, Infectiology and Metabolism, University of Marburg, Marburg, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Baumgart
authors.affiliation: Department of Gastroenterology and Gastrointestinal Oncology, University Medical Center Göttingen, Göttingen, Germany.
authors.forename: Nai-Ming
authors.initials: NM
authors.lastname: Chen
authors.affiliation: Schulze Center for Novel Therapeutics, Division of Oncology Research, Mayo Clinic, Rochester, Minnesota.
authors.forename: Jin-San
authors.initials: JS
authors.lastname: Zhang
authors.affiliation: Schulze Center for Novel Therapeutics, Division of Oncology Research, Mayo Clinic, Rochester, Minnesota.
authors.forename: Daniel D
authors.initials: DD
authors.lastname: Billadeau
authors.affiliation: Drug Discovery Program, Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago, Chicago, Illinois.
authors.forename: Irina N
authors.initials: IN
authors.lastname: Gaisina
authors.affiliation: Drug Discovery Program, Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago, Chicago, Illinois.
authors.forename: Alan P
authors.initials: AP
authors.lastname: Kozikowski
authors.affiliation: Barrow Brain Tumor Research Center, St. Joseph's Hospital and Medical Center, Phoenix, Arizona.
authors.forename: Shiv K
authors.initials: SK
authors.lastname: Singh
authors.affiliation: Department of Gastroenterology, Endocrinology, Infectiology and Metabolism, University of Marburg, Marburg, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Fink
authors.affiliation: Institute of Pathology, University Medical Center Göttingen, Göttingen, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Ströbel
authors.affiliation: Department of Gastroenterology, Endocrinology, Infectiology and Metabolism, University of Marburg, Marburg, Germany.
authors.forename: Caroline
authors.initials: C
authors.lastname: Klindt
authors.affiliation: Division of Anatomic Pathology, Mayo Clinic, Rochester, Minnesota.
authors.forename: Lizhi
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Division of Biostatistics, College of Medicine, Mayo Clinic, Rochester, Minnesota.
authors.forename: William R
authors.initials: WR
authors.lastname: Bamlet
authors.affiliation: Department of Gastroenterology and Gastrointestinal Oncology, University Medical Center Göttingen, Göttingen, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Koenig
authors.affiliation: Department of Gastroenterology and Gastrointestinal Oncology, University Medical Center Göttingen, Göttingen, Germany.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Hessmann
authors.affiliation: Department of Gastroenterology, Endocrinology, Infectiology and Metabolism, University of Marburg, Marburg, Germany.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Gress
authors.affiliation: Department of Gastroenterology and Gastrointestinal Oncology, University Medical Center Göttingen, Göttingen, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Ellenrieder
authors.affiliation: Department of Gastroenterology and Gastrointestinal Oncology, University Medical Center Göttingen, Göttingen, Germany. albrecht.neesse@med.uni-goettingen.de.
authors.forename: Albrecht
authors.initials: A
authors.lastname: Neesse
mesh_terms: D000818:Animals; D001665:Binding Sites; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D018450:Disease Progression; D015870:Gene Expression; D011905:Genes, ras; D000071679:Glycogen Synthase Kinase 3 beta / Q000378:metabolism*; D006801:Humans; D007249:Inflammation / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D046912:Multiprotein Complexes / Q000378:metabolism; D050778:NFATC Transcription Factors / Q000235:genetics / Q000378:metabolism*; D059372:Nucleotide Motifs; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D010766:Phosphorylation; D011485:Protein Binding; D055550:Protein Stability; D050796:STAT3 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D046912:Multiprotein Complexes; D050778:NFATC Transcription Factors; D050796:STAT3 Transcription Factor; D000071679:Glycogen Synthase Kinase 3 beta
references.citation: EMBO J. 2015 Feb 12;34(4):517-30
references.pmid: 25586376
references.citation: Neuroscience. 2010 Sep 1;169(3):1063-70
references.pmid: 20553816
references.citation: J Cell Sci. 2003 Apr 1;116(Pt 7):1175-86
references.pmid: 12615961
references.citation: Clin Cancer Res. 2006 Sep 1;12(17):5074-81
references.pmid: 16951223
references.citation: Cancer Discov. 2014 Jun;4(6):688-701
references.pmid: 24694735
references.citation: Gastroenterology. 2010 Mar;138(3):1189-99.e1-2
references.pmid: 19900447
references.citation: Nature. 2000 Jul 6;406(6791):86-90
references.pmid: 10894547
references.citation: Cancer Cell. 2011 Apr 12;19(4):456-69
references.pmid: 21481788
references.citation: J Immunol. 2011 Feb 1;186(3):1391-8
references.pmid: 21191064
references.citation: Med Oncol. 1998 Apr;15(1):32-6
references.pmid: 9643528
references.citation: Cancer Cell. 2007 Mar;11(3):291-302
references.pmid: 17349585
references.citation: Oncogene. 2011 Aug 25;30(34):3705-15
references.pmid: 21441955
references.citation: Cell Death Dis. 2014 Mar 27;5:e1142
references.pmid: 24675460
references.citation: Cancer Cell. 2013 Mar 18;23(3):406-20
references.pmid: 23453624
references.citation: Mol Cancer Ther. 2014 Feb;13(2):285-96
references.pmid: 24327518
references.citation: Cancer Cell. 2012 Jun 12;21(6):822-35
references.pmid: 22698406
references.citation: Cell. 2014 Jul 3;158(1):185-197
references.pmid: 24954535
references.citation: Cancer Res. 2005 Mar 15;65(6):2076-81
references.pmid: 15781615
references.citation: EMBO J. 2006 Aug 9;25(15):3714-24
references.pmid: 16874304
references.citation: Cancer Cell. 2011 Jun 14;19(6):728-39
references.pmid: 21665147
references.citation: Front Mol Neurosci. 2011 Nov 16;4:40
references.pmid: 22110425
references.citation: J Clin Invest. 2012 Feb;122(2):639-53
references.pmid: 22232209
references.citation: J Biol Chem. 2003 Aug 22;278(34):32227-35
references.pmid: 12794074
references.citation: Gastroenterology. 1998 Apr;114(4):859-61
references.pmid: 9547109
references.citation: CA Cancer J Clin. 2013 Jan;63(1):11-30
references.pmid: 23335087
references.citation: Cell. 2012 Apr 27;149(3):656-70
references.pmid: 22541435
references.citation: Cancer Res. 2011 Jul 15;71(14):5020-9
references.pmid: 21586612
references.citation: J Carcinog. 2012;11:15
references.pmid: 23230392
references.citation: J Biol Chem. 2011 Aug 19;286(33):28761-71
references.pmid: 21628454
references.citation: Mol Cancer Res. 2009 Mar;7(3):425-32
references.pmid: 19258413
references.citation: Cancer Discov. 2014 Apr;4(4):452-65
references.pmid: 24444711
references.citation: Cancer Cell. 2012 Sep 11;22(3):304-17
references.pmid: 22975374
references.citation: Cancer Cell. 2012 Jun 12;21(6):836-47
references.pmid: 22698407
references.citation: PLoS One. 2013;8(2):e55289
references.pmid: 23408967
references.citation: Proc Natl Acad Sci U S A. 2013 Jul 23;110(30):12325-30
references.pmid: 23836645
references.citation: Genes Dev. 2000 Aug 1;14(15):1837-51
references.pmid: 10921899
references.citation: Science. 1997 Mar 28;275(5308):1930-4
references.pmid: 9072970
references.citation: Gastroenterology. 2012 Feb;142(2):388-98.e1-7
references.pmid: 22079596
references.citation: J Biol Chem. 2001 Feb 2;276(5):3666-73
references.pmid: 11063740
references.citation: J Immunol. 1988 Apr 1;140(7):2243-8
references.pmid: 3258332
references.citation: Gastroenterology. 2003 Sep;125(3):891-905
references.pmid: 12949733
references.citation: Cancer Cell. 2012 Sep 11;22(3):318-30
references.pmid: 22975375
references.citation: Genes Dev. 1997 Apr 1;11(7):824-34
references.pmid: 9106655
references.citation: Cell. 1988 May 20;53(4):549-54
references.pmid: 2453289
references.citation: Cancer Discov. 2013 Jun;3(6):690-703
references.pmid: 23547054
references.citation: Int J Cancer. 2005 Jul 20;115(6):903-10
references.pmid: 15723355