Inhibition of KRAS-driven tumorigenicity by interruption of an autocrine cytokine circuit.

Although the roles of mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K) signaling in KRAS-driven tumorigenesis are well established, KRAS activates additional pathways required for tumor maintenance, the inhibition of which are likely to be necessary for effective KRAS-directed therapy. Here, we show that the IκB kinase (IKK)-related kinases Tank-binding kinase-1 (TBK1) and IKKε promote KRAS-driven tumorigenesis by regulating autocrine CCL5 and interleukin (IL)-6 and identify CYT387 as a potent JAK/TBK1/IKKε inhibitor. CYT387 treatment ablates RAS-associated cytokine signaling and impairs Kras-driven murine lung cancer growth. Combined CYT387 treatment and MAPK pathway inhibition induces regression of aggressive murine lung adenocarcinomas driven by Kras mutation and p53 loss. These observations reveal that TBK1/IKKε promote tumor survival by activating CCL5 and IL-6 and identify concurrent inhibition of TBK1/IKKε, Janus-activated kinase (JAK), and MEK signaling as an effective approach to inhibit the actions of oncogenic KRAS.

[1]  C. Der,et al.  Distinct requirements for Ras oncogenesis in human versus mouse cells. , 2002, Genes & development.

[2]  G. Condorelli,et al.  IκB kinase ε and TANK-binding kinase 1 activate AKT by direct phosphorylation , 2011, Proceedings of the National Academy of Sciences.

[3]  R. Weinberg,et al.  Species- and cell type-specific requirements for cellular transformation. , 2004, Cancer cell.

[4]  J. Cedarbaum.  Survival , 2004 .

[5]  Patricia Greninger,et al.  A gene expression signature associated with "K-Ras addiction" reveals regulators of EMT and tumor cell survival. , 2009, Cancer cell.

[6]  A. McMahon,et al.  The activity of Gli transcription factors is essential for Kras-induced pancreatic tumorigenesis , 2012, Proceedings of the National Academy of Sciences.

[7]  Ben S. Wittner,et al.  Systematic RNA interference reveals that oncogenic KRAS-driven cancers require TBK1 , 2009, Nature.

[8]  N. Hacohen,et al.  Highly parallel identification of essential genes in cancer cells , 2008, Proceedings of the National Academy of Sciences.

[9]  I. Hochberg,et al.  An inhibitor of the protein kinases TBK1/IKKε improves obesity-related metabolic dysfunctions , 2013, Nature Medicine.

[10]  K. Shokat,et al.  Targeting the cancer kinome through polypharmacology , 2010, Nature Reviews Cancer.

[11]  Bin Fang,et al.  Dissection of TBK1 signaling via phosphoproteomics in lung cancer cells , 2013, Proceedings of the National Academy of Sciences.

[12]  Julian Downward,et al.  The GATA2 Transcriptional Network Is Requisite for RAS Oncogene-Driven Non-Small Cell Lung Cancer , 2012, Cell.

[13]  J. Cheng,et al.  IKBKE is induced by STAT3 and tobacco carcinogen and determines chemosensitivity in non-small cell lung cancer , 2013, Oncogene.

[14]  Jonathan Melamed,et al.  Chemokine Signaling via the CXCR2 Receptor Reinforces Senescence , 2008, Cell.

[15]  W. Hahn,et al.  Emerging roles for the non-canonical IKKs in cancer , 2011, Oncogene.

[16]  M. Karin,et al.  Hepatic expression of HCV RNA-dependent RNA polymerase triggers innate immune signaling and cytokine production. , 2012, Molecular cell.

[17]  T. Jacks,et al.  Somatic activation of the K-ras oncogene causes early onset lung cancer in mice , 2001, Nature.

[18]  U. Brinkmann,et al.  Influence of TBK-1 on tumor angiogenesis and microvascular inflammation. , 2008, Frontiers in bioscience : a journal and virtual library.

[19]  Kate Owen,et al.  Author response: Registered report: COT drives resistance to RAF inhibition through MAP kinase pathway reactivation , 2015 .

[20]  K. Lim,et al.  Oncogenic Ras-induced secretion of IL6 is required for tumorigenesis. , 2007, Genes & development.

[21]  Andrew L. Kung,et al.  A murine lung cancer co-clinical trial identifies genetic modifiers of therapeutic response , 2012, Nature.

[22]  Ross Tubo,et al.  Mesenchymal stem cells within tumour stroma promote breast cancer metastasis , 2007, Nature.

[23]  D. Peeper,et al.  Oncogene-Induced Senescence Relayed by an Interleukin-Dependent Inflammatory Network , 2008, Cell.

[24]  R. Laborde,et al.  Safety and efficacy of CYT387, a JAK1 and JAK2 inhibitor, in myelofibrosis , 2013, Leukemia.

[25]  R. Collins.  Faculty Opinions recommendation of RalB GTPase-mediated activation of the IkappaB family kinase TBK1 couples innate immune signaling to tumor cell survival. , 2007 .

[26]  D. Levy,et al.  STAT3 plays a critical role in KRAS-induced pancreatic tumorigenesis. , 2011, Cancer research.

[27]  D. Feldser,et al.  Requirement for NF-κB signalling in a mouse model of lung adenocarcinoma , 2009, Nature.

[28]  B. Druker,et al.  CYT387, a novel JAK2 inhibitor, induces hematologic responses and normalizes inflammatory cytokines in murine myeloproliferative neoplasms. , 2010, Blood.

[29]  Shizuo Akira,et al.  Stat3/Socs3 activation by IL-6 transsignaling promotes progression of pancreatic intraepithelial neoplasia and development of pancreatic cancer. , 2011, Cancer cell.

[30]  T. Jacks,et al.  The differential effects of mutant p53 alleles on advanced murine lung cancer. , 2005, Cancer research.

[31]  M. White,et al.  Ral GTPases and cancer: linchpin support of the tumorigenic platform , 2008, Nature Reviews Cancer.

[32]  Gloria S. Huang,et al.  Abstract 5020: Potential lifestyle intervention in inner city women with endometrial cancer: A pilot study , 2011 .

[33]  A. Kimmelman,et al.  The dynamic nature of autophagy in cancer. , 2011, Genes & development.

[34]  J. Flores,et al.  The signaling adaptor p62 is an important NF-kappaB mediator in tumorigenesis. , 2008, Cancer cell.

[35]  W. Hahn,et al.  Structure and ubiquitination-dependent activation of TANK-binding kinase 1. , 2013, Cell reports.

[36]  J. Schornagel,et al.  Resistance to methotrexate due to gene amplification in a patient with acute leukemia. , 1984, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[37]  S. Akira,et al.  Stat3 and MMP7 contribute to pancreatic ductal adenocarcinoma initiation and progression. , 2011, Cancer cell.

[38]  Eric S. Lander,et al.  Integrative Genomic Approaches Identify IKBKE as a Breast Cancer Oncogene , 2007, Cell.

[39]  Ruiying Zhao,et al.  KrasG12D-induced IKK2/β/NF-κB activation by IL-1α and p62 feedforward loops is required for development of pancreatic ductal adenocarcinoma. , 2012, Cancer cell.

[40]  S. Akira,et al.  Activation of TBK1 and IKKε Kinases by Vesicular Stomatitis Virus Infection and the Role of Viral Ribonucleoprotein in the Development of Interferon Antiviral Immunity , 2004, Journal of Virology.

[41]  J. Minna,et al.  High expression of ligands for chemokine receptor CXCR2 in alveolar epithelial neoplasia induced by oncogenic kras. , 2006, Cancer research.

[42]  S. Wilkinson,et al.  TBK1 Kinase Addiction in Lung Cancer Cells Is Mediated via Autophagy of Tax1bp1/Ndp52 and Non-Canonical NF-κB Signalling , 2012, PloS one.

[43]  Marcin Piechota,et al.  Identification of interleukin-1 and interleukin-6-responsive genes in human monocyte-derived macrophages using microarrays. , 2008, Biochimica et biophysica acta.

[44]  Roger D Kamm,et al.  Screening therapeutic EMT blocking agents in a three-dimensional microenvironment. , 2013, Integrative biology : quantitative biosciences from nano to macro.

[45]  Eric Vangrevelinghe,et al.  Genetic resistance to JAK2 enzymatic inhibitors is overcome by HSP90 inhibition , 2011, The Journal of experimental medicine.

[46]  W. Hahn,et al.  Phosphorylation of the tumor suppressor CYLD by the breast cancer oncogene IKKepsilon promotes cell transformation. , 2009, Molecular cell.

[47]  M. White,et al.  RalB GTPase-Mediated Activation of the IκB Family Kinase TBK1 Couples Innate Immune Signaling to Tumor Cell Survival , 2006, Cell.

[48]  P. Cohen,et al.  Novel cross-talk within the IKK family controls innate immunity. , 2011, The Biochemical journal.

[49]  J. Downward.  Targeting RAS signalling pathways in cancer therapy , 2003, Nature Reviews Cancer.

[50]  C. Y. Wang,et al.  Requirement of NF-kappaB activation to suppress p53-independent apoptosis induced by oncogenic Ras. , 1997, Science.

[51]  D. Bar-Sagi,et al.  Ras-induced interleukin-8 expression plays a critical role in tumor growth and angiogenesis. , 2004, Cancer cell.

[52]  M. White,et al.  TBK1 directly engages Akt/PKB survival signaling to support oncogenic transformation. , 2011, Molecular cell.