c-Myc is an important direct target of Notch1 in T-cell acute lymphoblastic leukemia/lymphoma.

Human acute T-cell lymphoblastic leukemias and lymphomas (T-ALL) are commonly associated with gain-of-function mutations in Notch1 that contribute to T-ALL induction and maintenance. Starting from an expression-profiling screen, we identified c-myc as a direct target of Notch1 in Notch-dependent T-ALL cell lines, in which Notch accounts for the majority of c-myc expression. In functional assays, inhibitors of c-myc interfere with the progrowth effects of activated Notch1, and enforced expression of c-myc rescues multiple Notch1-dependent T-ALL cell lines from Notch withdrawal. The existence of a Notch1-c-myc signaling axis was bolstered further by experiments using c-myc-dependent murine T-ALL cells, which are rescued from withdrawal of c-myc by retroviral transduction of activated Notch1. This Notch1-mediated rescue is associated with the up-regulation of endogenous murine c-myc and its downstream transcriptional targets, and the acquisition of sensitivity to Notch pathway inhibitors. Additionally, we show that primary murine thymocytes at the DN3 stage of development depend on ligand-induced Notch signaling to maintain c-myc expression. Together, these data implicate c-myc as a developmentally regulated direct downstream target of Notch1 that contributes to the growth of T-ALL cells.

[1]  S. Artavanis-Tsakonas,et al.  The molecular genetics of Enhancer of split, a gene required for embryonic neural development in Drosophila. , 1988, The EMBO journal.

[2]  J. Sklar,et al.  TAN-1, the human homolog of the Drosophila Notch gene, is broken by chromosomal translocations in T lymphoblastic neoplasms , 1991, Cell.

[3]  Multiple mechanisms of tumorigenesis in E mu-myc transgenic mice. , 1993, Cancer research.

[4]  A. Sharpe,et al.  The ikaros gene is required for the development of all lymphoid lineages , 1994, Cell.

[5]  K. Georgopoulos,et al.  The Ikaros gene encodes a family of functionally diverse zinc finger DNA-binding proteins , 1994, Molecular and cellular biology.

[6]  T. Honjo,et al.  Recognition sequence of a highly conserved DNA binding protein RBP-Jx , 1994 .

[7]  K. Georgopoulos,et al.  A dominant mutation in the Ikaros gene leads to rapid development of leukemia and lymphoma , 1995, Cell.

[8]  Christel Brou,et al.  Signalling downstream of activated mammalian Notch , 1995, Nature.

[9]  J. Sklar,et al.  Exclusive development of T cell neoplasms in mice transplanted with bone marrow expressing activated Notch alleles , 1996, The Journal of experimental medicine.

[10]  H. Weintraub,et al.  Signal transduction by activated mNotch: importance of proteolytic processing and its regulation by the extracellular domain. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[11]  L. Girard,et al.  Frequent provirus insertional mutagenesis of Notch1 in thymomas of MMTVD/myc transgenic mice suggests a collaboration of c-myc and Notch1 for oncogenesis. , 1996, Genes & development.

[12]  C. Vinson,et al.  A general method to design dominant negatives to B-HLHZip proteins that abolish DNA binding. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[13]  T. Schedl,et al.  Germ-line tumor formation caused by activation of glp-1, a Caenorhabditis elegans member of the Notch family of receptors. , 1997, Development.

[14]  Raphael Kopan,et al.  Notch-1 signalling requires ligand-induced proteolytic release of intracellular domain , 1998, Nature.

[15]  S. Artavanis-Tsakonas,et al.  Notch Signaling : Cell Fate Control and Signal Integration in Development , 1999 .

[16]  Li Wu,et al.  Pre–T Cell Receptor (Tcr) and Tcr-Controlled Checkpoints in T Cell Differentiation Are Set by Ikaros , 1999, The Journal of experimental medicine.

[17]  D. Prober,et al.  Drosophila myc Regulates Cellular Growth during Development , 1999, Cell.

[18]  J. Aster,et al.  Notch1 expression in early lymphopoiesis influences B versus T lineage determination. , 1999, Immunity.

[19]  Iva Greenwald,et al.  Presenilin is required for activity and nuclear access of Notch in Drosophila , 1999, Nature.

[20]  William J. Ray,et al.  A presenilin-1-dependent γ-secretase-like protease mediates release of Notch intracellular domain , 1999, Nature.

[21]  D. Felsher,et al.  Reversible tumorigenesis by MYC in hematopoietic lineages. , 1999, Molecular cell.

[22]  U. Weidle,et al.  Control of cell growth by c-Myc in the absence of cell division , 1999, Current Biology.

[23]  M. Fortini,et al.  Neurogenic phenotypes and altered Notch processing in Drosophila Presenilin mutants , 1999, Nature.

[24]  E. Lai,et al.  Discrete enhancer elements mediate selective responsiveness of enhancer of split complex genes to common transcriptional activators. , 1999, Developmental biology.

[25]  H. Macdonald,et al.  Deficient T cell fate specification in mice with an induced inactivation of Notch1. , 1999, Immunity.

[26]  J. Kimble,et al.  LAG-3 is a putative transcriptional activator in the C. elegans Notch pathway , 2000, Nature.

[27]  J. Kimble,et al.  Mastermind is a putative activator for Notch , 2000, Current Biology.

[28]  J. Aster,et al.  MAML 1 , a human homologue of Drosophila Mastermind , is a transcriptional coactivator for NOTCH receptors , 2000 .

[29]  I. Bernstein,et al.  Pluripotent, cytokine-dependent, hematopoietic stem cells are immortalized by constitutive Notch1 signaling , 2000, Nature Medicine.

[30]  L. Girard,et al.  Two Distinct Notch1 Mutant Alleles Are Involved in the Induction of T-Cell Leukemia in c-myc Transgenic Mice , 2000, Molecular and Cellular Biology.

[31]  A Cumano,et al.  A novel proteolytic cleavage involved in Notch signaling: the role of the disintegrin-metalloprotease TACE. , 2000, Molecular cell.

[32]  Jon C. Aster,et al.  Essential Roles for Ankyrin Repeat and Transactivation Domains in Induction of T-Cell Leukemia by Notch1 , 2000, Molecular and Cellular Biology.

[33]  M. Bevan,et al.  Notch1 signaling promotes the maturation of CD4 and CD8 SP thymocytes. , 2000, Immunity.

[34]  R. Eisenman,et al.  The Myc/Max/Mad network and the transcriptional control of cell behavior. , 2000, Annual review of cell and developmental biology.

[35]  T. Hampton,et al.  A carboxy-terminal deletion mutant of Notch1 accelerates lymphoid oncogenesis in E2A-PBX1 transgenic mice , 2000 .

[36]  Raphael Kopan,et al.  A ligand-induced extracellular cleavage regulates gamma-secretase-like proteolytic activation of Notch1. , 2000, Molecular cell.

[37]  James D. Griffin,et al.  MAML1, a human homologue of Drosophila Mastermind, is a transcriptional co-activator for NOTCH receptors , 2000, Nature Genetics.

[38]  U. Lendahl,et al.  Constitutive activation of NF‐κB and T‐cell leukemia/lymphoma in Notch3 transgenic mice , 2000, The EMBO journal.

[39]  A. Capobianco,et al.  Induction of Cyclin D1 Transcription and CDK2 Activity by Notchic: Implication for Cell Cycle Disruption in Transformation by Notchic , 2001, Molecular and Cellular Biology.

[40]  A. Hayday,et al.  Defining the specific physiological requirements for c-Myc in T cell development , 2001, Nature Immunology.

[41]  C. Li,et al.  Model-based analysis of oligonucleotide arrays: expression index computation and outlier detection. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[42]  Todd,et al.  Diffuse large B-cell lymphoma outcome prediction by gene-expression profiling and supervised machine learning , 2002, Nature Medicine.

[43]  Boris Reizis,et al.  Direct induction of T lymphocyte-specific gene expression by the mammalian Notch signaling pathway. , 2002, Genes & development.

[44]  Carla Grandori,et al.  Modulation of T‐lymphocyte development, growth and cell size by the Myc antagonist and transcriptional repressor Mad1 , 2002, The EMBO journal.

[45]  E. Lander,et al.  Gene expression signatures define novel oncogenic pathways in T cell acute lymphoblastic leukemia. , 2002, Cancer cell.

[46]  H. Macdonald,et al.  Inactivation of Notch1 impairs VDJbeta rearrangement and allows pre-TCR-independent survival of early alpha beta Lineage Thymocytes. , 2002, Immunity.

[47]  A. Ferrando,et al.  Requirement for cyclin D3 in lymphocyte development and T cell leukemias. , 2003, Cancer cell.

[48]  E. Macintyre,et al.  Analysis of TCR, pT alpha, and RAG-1 in T-acute lymphoblastic leukemias improves understanding of early human T-lymphoid lineage commitment. , 2003, Blood.

[49]  Andrew P. Weng,et al.  Structural Requirements for Assembly of the CSL·Intracellular Notch1·Mastermind-like 1 Transcriptional Activation Complex* , 2003, Journal of Biological Chemistry.

[50]  D. Scadden,et al.  Osteoblastic cells regulate the haematopoietic stem cell niche , 2003, Nature.

[51]  D. Selkoe,et al.  Notch and the amyloid precursor protein are cleaved by similar gamma-secretase(s). , 2003, Biochemistry.

[52]  R. Eisenman,et al.  Direct Regulation of RNA Polymerase III Transcription by RB, p53 and c-Myc , 2003, Cell cycle.

[53]  James D. Griffin,et al.  Growth Suppression of Pre-T Acute Lymphoblastic Leukemia Cells by Inhibition of Notch Signaling , 2003, Molecular and Cellular Biology.

[54]  David M Langenau,et al.  Myc-Induced T Cell Leukemia in Transgenic Zebrafish , 2003, Science.

[55]  A. Capobianco,et al.  Perturbation of Ikaros isoform selection by MLV integration is a cooperative event in Notch(IC)-induced T cell leukemogenesis. , 2003, Cancer cell.

[56]  Andrew P. Weng,et al.  Activating Mutations of NOTCH1 in Human T Cell Acute Lymphoblastic Leukemia , 2004, Science.

[57]  Richard A Flavell,et al.  Instruction of Distinct CD4 T Helper Cell Fates by Different Notch Ligands on Antigen-Presenting Cells , 2004, Cell.

[58]  J. Aster,et al.  Notch Subunit Heterodimerization and Prevention of Ligand-Independent Proteolytic Activation Depend, Respectively, on a Novel Domain and the LNR Repeats , 2004, Molecular and Cellular Biology.

[59]  Thomas M. Schmitt,et al.  Induction of T cell development and establishment of T cell competence from embryonic stem cells differentiated in vitro , 2004, Nature Immunology.

[60]  Freddy Radtke,et al.  Notch regulation of lymphocyte development and function , 2004, Nature Immunology.

[61]  M. Kubo,et al.  Regulation of αβ/γδ T cell lineage commitment and peripheral T cell responses by Notch/RBP-J signaling , 2004 .

[62]  I. Matsumura,et al.  Roles for c-Myc in Self-renewal of Hematopoietic Stem Cells* , 2004, Journal of Biological Chemistry.

[63]  C. Guidos,et al.  Requirement for Notch1 signals at sequential early stages of intrathymic T cell development , 2005, Nature Immunology.

[64]  W. Pear,et al.  Notch signaling is an important regulator of type 2 immunity , 2005, The Journal of experimental medicine.

[65]  M. Caudy,et al.  A DNA Transcription Code for Cell-Specific Gene Activation by Notch Signaling , 2005, Current Biology.

[66]  T. Südhof,et al.  Nicastrin Functions as a γ-Secretase-Substrate Receptor , 2005, Cell.

[67]  B. Edgar,et al.  Myc-dependent regulation of ribosomal RNA synthesis during Drosophila development , 2005, Nature Cell Biology.

[68]  Lars-Gunnar Larsson,et al.  c-Myc associates with ribosomal DNA and activates RNA polymerase I transcription , 2005, Nature Cell Biology.

[69]  Tannishtha Reya,et al.  Integration of Notch and Wnt signaling in hematopoietic stem cell maintenance , 2005, Nature Immunology.

[70]  W. Pear,et al.  Regulation of lymphoid development, differentiation, and function by the Notch pathway. , 2005, Annual review of immunology.

[71]  A. Trumpp,et al.  The Myc trilogy: lord of RNA polymerases , 2005, Nature Cell Biology.

[72]  Carla Grandori,et al.  c-Myc binds to human ribosomal DNA and stimulates transcription of rRNA genes by RNA polymerase I , 2005, Nature Cell Biology.

[73]  J. Aster,et al.  Notch signaling controls the generation and differentiation of early T lineage progenitors , 2005, Nature Immunology.

[74]  M. Ciofani,et al.  Notch promotes survival of pre–T cells at the β-selection checkpoint by regulating cellular metabolism , 2005, Nature Immunology.

[75]  T. Golde,et al.  Inhibitors of γ-secretase block in vivo and in vitro T helper type 1 polarization by preventing Notch upregulation of Tbx21 , 2005, Nature Immunology.

[76]  J. Letterio,et al.  Notch1 mutations are important for leukemic transformation in murine models of precursor-T leukemia/lymphoma. , 2006, Blood.

[77]  P. Kastner,et al.  Notch Activation Is an Early and Critical Event during T-Cell Leukemogenesis in Ikaros-Deficient Mice , 2006, Molecular and Cellular Biology.

[78]  J. Aster,et al.  Leukemia-Associated Mutations within the NOTCH1 Heterodimerization Domain Fall into at Least Two Distinct Mechanistic Classes , 2006, Molecular and Cellular Biology.

[79]  E. Rothenberg,et al.  Developmental and Molecular Characterization of Emerging β- and γδ-Selected Pre-T Cells in the Adult Mouse Thymus , 2006 .

[80]  F. Alt,et al.  Activating Notch1 mutations in mouse models of T-ALL. , 2005, Blood.

[81]  G. Stormo,et al.  Target Selectivity of Vertebrate Notch Proteins , 2006, Journal of Biological Chemistry.

[82]  M. Greenberg,et al.  Identification of Newly Transcribed RNA , 1994, Current protocols in molecular biology.