JAK2-V617F-induced MAPK activity is regulated by PI3K and acts synergistically with PI3K on the proliferation of JAK2-V617F-positive cells

The identification of a constitutively active JAK2 mutant, namely JAK2-V617F, was a milestone in the understanding of Philadelphia chromosome-negative myeloproliferative neoplasms. The JAK2-V617F mutation confers cytokine hypersensitivity, constitutive activation of the JAK-STAT pathway, and cytokine-independent growth. In this study we investigated the mechanism of JAK2-V617F-dependent signaling with a special focus on the activation of the MAPK pathway. We observed JAK2-V617F-dependent deregulated activation of the multi-site docking protein Gab1 as indicated by constitutive, PI3K-dependent membrane localization and tyrosine phosphorylation of Gab1. Furthermore, we demonstrate that PI3K signaling regulates MAPK activation in JAK2-V617F-positve cells. This cross-regulation of the MAPK pathway by PI3K affects JAK2-V617F-specific target gene induction, erythroid colony formation, and regulates proliferation of JAK2-V617F-positive patient cells in a synergistically manner.

[1]  David R. Croucher,et al.  Functional characterization of cancer-associated Gab1 mutations , 2013, Oncogene.

[2]  P. Guglielmelli,et al.  mTOR Inhibitors Alone and in Combination with JAK2 Inhibitors Effectively Inhibit Cells of Myeloproliferative Neoplasms , 2013, PloS one.

[3]  I. Behrmann,et al.  Cooperative effects of Janus and Aurora kinase inhibition by CEP701 in cells expressing Jak2V617F , 2013, Journal of cellular and molecular medicine.

[4]  K. Bhalla,et al.  Role of additional novel therapies in myeloproliferative neoplasms. , 2012, Hematology/oncology clinics of North America.

[5]  A. Mead,et al.  Guideline for the diagnosis and management of myelofibrosis , 2012, British journal of haematology.

[6]  Caleb B. McDonald,et al.  Bivalent binding drives the formation of the Grb2–Gab1 signaling complex in a noncooperative manner , 2012, The FEBS journal.

[7]  Anna Dittrich,et al.  Interleukin-6 signalling: more than Jaks and STATs. , 2012, European journal of cell biology.

[8]  S. Verstovsek,et al.  Janus kinase Inhibition and its effect upon the therapeutic landscape for myelofibrosis: from palliation to cure? , 2012, British journal of haematology.

[9]  Jason Gotlib,et al.  A double-blind, placebo-controlled trial of ruxolitinib for myelofibrosis. , 2012, The New England journal of medicine.

[10]  Francisco Cervantes,et al.  JAK inhibition with ruxolitinib versus best available therapy for myelofibrosis. , 2012, The New England journal of medicine.

[11]  M. Karin,et al.  TANK-binding kinase 1 (TBK1) controls cell survival through PAI-2/serpinB2 and transglutaminase 2 , 2011, Proceedings of the National Academy of Sciences.

[12]  A. Vannucchi.  Management of myelofibrosis. , 2011, Hematology. American Society of Hematology. Education Program.

[13]  K. Bhalla,et al.  Heat Shock Protein 90 Inhibitor Is Synergistic with JAK2 Inhibitor and Overcomes Resistance to JAK2-TKI in Human Myeloproliferative Neoplasm Cells , 2011, Clinical Cancer Research.

[14]  A. Vannucchi,et al.  Epigenetics and mutations in chronic myeloproliferative neoplasms , 2011, Haematologica.

[15]  G. Giaccone,et al.  American Society of Clinical Oncology provisional clinical opinion: epidermal growth factor receptor (EGFR) Mutation testing for patients with advanced non-small-cell lung cancer considering first-line EGFR tyrosine kinase inhibitor therapy. , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[16]  Catriona Jamieson,et al.  Safety and efficacy of TG101348, a selective JAK2 inhibitor, in myelofibrosis. , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[17]  D. Lipka,et al.  FLT3 as a therapeutic target in AML: still challenging after all these years. , 2010, Blood.

[18]  K. Ross,et al.  HSP90 is a therapeutic target in JAK2-dependent myeloproliferative neoplasms in mice and humans. , 2010, The Journal of clinical investigation.

[19]  Ayalew Tefferi,et al.  Safety and efficacy of INCB018424, a JAK1 and JAK2 inhibitor, in myelofibrosis. , 2010, The New England journal of medicine.

[20]  I. Behrmann,et al.  Perspectives for the use of structural information and chemical genetics to develop inhibitors of Janus kinases , 2010, Journal of cellular and molecular medicine.

[21]  Roger J Daly,et al.  Function, regulation and pathological roles of the Gab/DOS docking proteins , 2009, Cell Communication and Signaling.

[22]  I. Behrmann,et al.  SOCS-mediated downregulation of mutant Jak2 (V617F, T875N and K539L) counteracts cytokine-independent signaling , 2009, Oncogene.

[23]  M. Mann,et al.  Lysine Acetylation Targets Protein Complexes and Co-Regulates Major Cellular Functions , 2009, Science.

[24]  J. Prchal,et al.  Erythropoiesis in polycythemia vera is hyper-proliferative and has accelerated maturation. , 2009, Blood cells, molecules & diseases.

[25]  M. Lewitzky,et al.  Distinct binding modes of two epitopes in Gab2 that interact with the SH3C domain of Grb2. , 2009, Structure.

[26]  J. Radich,et al.  Six-year follow-up of patients receiving imatinib for the first-line treatment of chronic myeloid leukemia , 2009, Leukemia.

[27]  F. Schaper,et al.  A new mechanism for the regulation of Gab1 recruitment to the plasma membrane , 2009, Journal of Cell Science.

[28]  S. Paggi,et al.  Sorafenib in Advanced Hepatocellular Carcinoma , 2008 .

[29]  F. Schaper,et al.  A novel mechanism for the regulation of Gab1 recruitment to the plasma membrane , 2008, Journal of Cell Science.

[30]  A. Suhrbier,et al.  SerpinB2 protection of retinoblastoma protein from calpain enhances tumor cell survival. , 2008, Cancer research.

[31]  C. Pecquet,et al.  Mining for JAK-STAT mutations in cancer. , 2008, Trends in biochemical sciences.

[32]  Ting-Chao Chou,et al.  Preclinical versus clinical drug combination studies , 2008, Leukemia & lymphoma.

[33]  R. Tiedt,et al.  Ratio of mutant JAK2-V617F to wild-type Jak2 determines the MPD phenotypes in transgenic mice. , 2007, Blood.

[34]  J. Mendell,et al.  Regulated expression of microRNAs in normal and polycythemia vera erythropoiesis. , 2007, Experimental hematology.

[35]  J. Prchal,et al.  In vitro expansion of erythroid progenitors from polycythemia vera patients leads to decrease in JAK2 V617F allele. , 2007, Experimental hematology.

[36]  M. Stratton,et al.  JAK2 exon 12 mutations in polycythemia vera and idiopathic erythrocytosis. , 2007, The New England journal of medicine.

[37]  I. Behrmann,et al.  A cost effective non-commercial ECL-solution for Western blot detections yielding strong signals and low background. , 2007, Journal of immunological methods.

[38]  R. Pazdur,et al.  Sorafenib for the Treatment of Advanced Renal Cell Carcinoma , 2006, Clinical Cancer Research.

[39]  Sandra A. Moore,et al.  JAK2T875N is a novel activating mutation that results in myeloproliferative disease with features of megakaryoblastic leukemia in a murine bone marrow transplantation model. , 2006, Blood.

[40]  D. Steensma.  JAK2 V617F in myeloid disorders: molecular diagnostic techniques and their clinical utility: a paper from the 2005 William Beaumont Hospital Symposium on Molecular Pathology. , 2006, The Journal of molecular diagnostics : JMD.

[41]  Ting-Chao Chou,et al.  Theoretical Basis, Experimental Design, and Computerized Simulation of Synergism and Antagonism in Drug Combination Studies , 2006, Pharmacological Reviews.

[42]  L. Hawthorn,et al.  Induction of the plasminogen activator inhibitor-2 in cells expressing the ZNF198/FGFR1 fusion kinase that is involved in atypical myeloproliferative disease. , 2006, Blood.

[43]  H. Lodish,et al.  Expression of a homodimeric type I cytokine receptor is required for JAK2V617F-mediated transformation. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[44]  E. Seto,et al.  Acetylation and deacetylation of non-histone proteins. , 2005, Gene.

[45]  D. Oscier,et al.  Widespread occurrence of the JAK2 V617F mutation in chronic myeloproliferative disorders. , 2005, Blood.

[46]  Qingshan Li,et al.  Identification of an Acquired JAK2 Mutation in Polycythemia Vera* , 2005, Journal of Biological Chemistry.

[47]  R. Hoffman,et al.  Constitutive mobilization of CD34+ cells into the peripheral blood in idiopathic myelofibrosis may be due to the action of a number of proteases. , 2005, Blood.

[48]  Mario Cazzola,et al.  A gain-of-function mutation of JAK2 in myeloproliferative disorders. , 2005, The New England journal of medicine.

[49]  Stefan N. Constantinescu,et al.  A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera , 2005, Nature.

[50]  Sandra A. Moore,et al.  Activating mutation in the tyrosine kinase JAK2 in polycythemia vera, essential thrombocythemia, and myeloid metaplasia with myelofibrosis. , 2005, Cancer cell.

[51]  P. Campbell,et al.  Acquired mutation of the tyrosine kinase JAK2 in human myeloproliferative disorders , 2005, The Lancet.

[52]  T. Alain,et al.  Stromelysin-2 (Matrix Metalloproteinase 10) Is Inducible in Lymphoma Cells and Accelerates the Growth of Lymphoid Tumors In Vivo1 , 2004, The Journal of Immunology.

[53]  W. Vainchenker,et al.  Multiple signaling pathways are involved in erythropoietin-independent differentiation of erythroid progenitors in polycythemia vera. , 2004, Experimental hematology.

[54]  N. Brünner,et al.  Elevated plasma levels of TIMP‐1 correlate with plasma suPAR/uPA in patients with chronic myeloproliferative disorders , 2003, European journal of haematology.

[55]  N. Brünner,et al.  Collagen metabolism and enzymes of the urokinase plasminogen activator system in chronic myeloproliferative disorders: correlation between plasma‐soluble urokinase plasminogen activator receptor and serum markers for collagen metabolism , 2003, European journal of haematology.

[56]  Rafael A Irizarry,et al.  Exploration, normalization, and summaries of high density oligonucleotide array probe level data. , 2003, Biostatistics.

[57]  B. Neel,et al.  The "Gab" in signal transduction. , 2003, Trends in cell biology.

[58]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

[59]  L. Pechet,et al.  Clinical Implications of Elevated PAI-1 Revisited: Multiple Arterial Thrombosis in a Patient with Essential Thrombocythemia and Elevated Plasminogen Activator Inhibitor-1 (PAI-1) Levels with a Review of the Literature , 1999, Journal of Thrombosis and Thrombolysis.

[60]  B. Telek,et al.  Reduced in vitro clot lysis and release of more active platelet PAI-1 in polycythemia vera and essential thrombocythemia. , 1998, Thrombosis research.

[61]  J. Cancelas,et al.  High plasma levels of plasminogen activator inhibitor 1 (PAI-1) in polycythemia vera and essential thrombocythemia are associated with thrombosis. , 1994, Thrombosis research.

[62]  Prchal Jf,et al.  Letter: Bone-marrow responses in polycythemia vera. , 1974 .

[63]  Kevin D Bunting,et al.  Grb2-associated binding (Gab) proteins in hematopoietic and immune cell biology. , 2011, American journal of blood research.

[64]  A. Tefferi.  Classification, diagnosis and management of myeloproliferative disorders in the JAK2V617F era. , 2006, Hematology. American Society of Hematology. Education Program.

[65]  B. Jenkins,et al.  Acquiring signalling specificity from the cytokine receptor gp130. , 2004, Trends in genetics : TIG.

[66]  L. Pechet,et al.  Clinical implications of elevated PAI-1 revisited: multiple arterial thrombosis in a patient with essential thrombocythemia and elevated plasminogen activator inhibitor-1 (PAI-1) levels: a case report and review of the literature. , 1999, Journal of thrombosis and thrombolysis.

[67]  J. Massagué.  TGF-beta signal transduction. , 1998, Annual review of biochemistry.

[68]  T. Chou,et al.  Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. , 1984, Advances in enzyme regulation.

[69]  A. Axelrad,et al.  Letter: Bone-marrow responses in polycythemia vera. , 1974, The New England journal of medicine.