CSF3R T618I Collaborates With RUNX1-RUNX1T1 to Expand Hematopoietic Progenitors and Sensitizes to GLI Inhibition

Activating colony-stimulating factor-3 receptor gene (CSF3R) mutations are recurrent in acute myeloid leukemia (AML) with t(8;21) translocation. However, the nature of oncogenic collaboration between alterations of CSF3R and the t(8;21) associated RUNX1-RUNX1T1 fusion remains unclear. In CD34+ hematopoietic stem and progenitor cells from healthy donors, double oncogene expression led to a clonal advantage, increased self-renewal potential, and blast-like morphology and distinct immunophenotype. Gene expression profiling revealed hedgehog signaling as a potential mechanism, with upregulation of GLI2 constituting a putative pharmacological target. Both primary hematopoietic cells and the t(8;21) positive AML cell line SKNO-1 showed increased sensitivity to the GLI inhibitor GANT61 when expressing CSF3R T618I. Our findings suggest that during leukemogenesis, the RUNX1-RUNXT1 fusion and CSF3R mutation act in a synergistic manner to alter hedgehog signaling, which can be exploited therapeutically.

[1]  J. Park,et al.  Cross-tissue immune cell analysis reveals tissue-specific features in humans , 2022, Science.

[2]  Lucas E. Wange,et al.  Prime-seq, efficient and powerful bulk RNA sequencing , 2021, bioRxiv.

[3]  Eunjung Kim,et al.  Activity-Based Protein Profiling Reveals Potential Dasatinib Targets in Gastric Cancer , 2020, International journal of molecular sciences.

[4]  L. Møller,et al.  Crosstalk of Hedgehog and mTORC1 Pathways , 2020, Cells.

[5]  G. Martinelli,et al.  Hedgehog Pathway Inhibitors: A New Therapeutic Class for the Treatment of Acute Myeloid Leukemia. , 2020, Blood cancer discovery.

[6]  R. Ries,et al.  Prognostic Impact of CSF3R Mutations in Favorable Risk Childhood Acute Myeloid Leukemia. , 2020, Blood.

[7]  Johannes W. Bagnoli,et al.  ZBTB7A prevents RUNX1-RUNX1T1-dependent clonal expansion of human hematopoietic stem and progenitor cells , 2020, Oncogene.

[8]  W. Hiddemann,et al.  The clinical mutatome of core binding factor leukemia , 2020, Leukemia.

[9]  L. Bullinger,et al.  Genomic landscape and clonal evolution of acute myeloid leukemia with t(8;21): an international study on 331 patients. , 2019, Blood.

[10]  S. Mori,et al.  Functional Genome-wide Screening Identifies Targets and Pathways Sensitizing Pancreatic Cancer Cells to Dasatinib , 2018, Journal of Cancer.

[11]  E. Hoster,et al.  Genetic heterogeneity of cytogenetically normal AML with mutations of CEBPA. , 2018, Blood advances.

[12]  Lucas E. Wange,et al.  Sensitive and powerful single-cell RNA sequencing using mcSCRB-seq , 2018, Nature Communications.

[13]  F. Wang,et al.  CSF3R Mutations are frequently associated with abnormalities of RUNX1, CBFB, CEBPA, and NPM1 genes in acute myeloid leukemia , 2018, Cancer.

[14]  J. Tyner,et al.  Gain-of-function mutations in granulocyte colony–stimulating factor receptor (CSF3R) reveal distinct mechanisms of CSF3R activation , 2018, The Journal of Biological Chemistry.

[15]  Fabian J Theis,et al.  SCANPY: large-scale single-cell gene expression data analysis , 2018, Genome Biology.

[16]  S. Meshinchi,et al.  CSF3R Mutations Synergize with CEBPA and SETBP1 Mutations in Acute Myeloid Leukemia and Chronic Neutrophilic Leukemia , 2017 .

[17]  J. Tyner,et al.  Genomics of chronic neutrophilic leukemia. , 2017, Blood.

[18]  G. Boucher,et al.  Chemo-genomic interrogation of CEBPA mutated AML reveals recurrent CSF3R mutations and subgroup sensitivity to JAK inhibitors. , 2016, Blood.

[19]  Jason E. Farrar,et al.  CSF3R mutations have a high degree of overlap with CEBPA mutations in pediatric AML. , 2016, Blood.

[20]  Feimeng Zheng,et al.  Targeting GLI1 Suppresses Cell Growth and Enhances Chemosensitivity in CD34+ Enriched Acute Myeloid Leukemia Progenitor Cells , 2016, Cellular Physiology and Biochemistry.

[21]  D. Grimwade,et al.  Molecular landscape of acute myeloid leukemia in younger adults and its clinical relevance. , 2016, Blood.

[22]  L. Marchionni,et al.  Integration of Hedgehog and mutant FLT3 signaling in myeloid leukemia , 2015, Science Translational Medicine.

[23]  Richard A. Moore,et al.  GLI2 inhibition abrogates human leukemia stem cell dormancy , 2015, Journal of Translational Medicine.

[24]  K. Wagner,et al.  Expression of Hedgehog Pathway Mediator GLI Represents a Negative Prognostic Marker in Human Acute Myeloid Leukemia and Its Inhibition Exerts Antileukemic Effects , 2015, Clinical Cancer Research.

[25]  R. Henschler,et al.  Activating c-KIT mutations confer oncogenic cooperativity and rescue RUNX1/ETO-induced DNA damage and apoptosis in human primary CD34+ hematopoietic progenitors , 2014, Leukemia.

[26]  B. Druker,et al.  Ligand Independence of the T618I Mutation in the Colony-stimulating Factor 3 Receptor (CSF3R) Protein Results from Loss of O-Linked Glycosylation and Increased Receptor Dimerization* , 2014, The Journal of Biological Chemistry.

[27]  Angela G. Fleischman,et al.  CSF3R T618I Mouse Bone Marrow Transplant Model Of Neutrophilic Leukemia , 2013 .

[28]  Angela G. Fleischman,et al.  Oncogenic CSF3R mutations in chronic neutrophilic leukemia and atypical CML. , 2013, The New England journal of medicine.

[29]  Deng Pan,et al.  Gli inhibitor GANT61 causes apoptosis in myeloid leukemia cells and acts in synergy with rapamycin. , 2012, Leukemia research.

[30]  S. Okabe,et al.  Effects of the hedgehog inhibitor GDC-0449, alone or in combination with dasatinib, on BCR-ABL-positive leukemia cells. , 2012, Stem cells and development.

[31]  R. Norton,et al.  Suppression of cytokine signaling by SOCS3: characterization of the mode of inhibition and the basis of its specificity. , 2012, Immunity.

[32]  E. Vellenga,et al.  Dasatinib impairs long-term expansion of leukemic progenitors in a subset of acute myeloid leukemia cases , 2010, Annals of Hematology.

[33]  Bin Fang,et al.  A chemical and phosphoproteomic characterization of dasatinib action in lung cancer , 2010, Nature chemical biology.

[34]  Gerhard Dürnberger,et al.  Chemical proteomic profiles of the BCR-ABL inhibitors imatinib, nilotinib, and dasatinib reveal novel kinase and nonkinase targets. , 2007, Blood.

[35]  W. Hiddemann,et al.  AML1–ETO meets JAK2: clinical evidence for the two hit model of leukemogenesis from a myeloproliferative syndrome progressing to acute myeloid leukemia , 2007, Leukemia.

[36]  W. Alexander,et al.  The SOCS box of suppressor of cytokine signaling-3 contributes to the control of G-CSF responsiveness in vivo. , 2007, Blood.

[37]  Bernhard Kuster,et al.  Quantitative chemical proteomics reveals mechanisms of action of clinical ABL kinase inhibitors , 2007, Nature Biotechnology.

[38]  S. H. Lee,et al.  The JAK2 V617F mutation in de novo acute myelogenous leukemias , 2006, Oncogene.

[39]  Hui Zhao,et al.  AML1-ETO fusion protein up-regulates TRKA mRNA expression in human CD34+ cells, allowing nerve growth factor-induced expansion. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[40]  L. Peterson,et al.  The 8;21 translocation in leukemogenesis , 2004, Oncogene.

[41]  S. Jhanwar,et al.  Maintaining the self-renewal and differentiation potential of human CD34+ hematopoietic cells using a single genetic element. , 2003, Blood.

[42]  D. Gilliland,et al.  Genetics of myeloid leukemias. , 2003, Annual review of genomics and human genetics.

[43]  M. Tomasson,et al.  An activated receptor tyrosine kinase, TEL/PDGFβR, cooperates with AML1/ETO to induce acute myeloid leukemia in mice , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[44]  J. Downing,et al.  The AML1-ETO fusion gene promotes extensive self-renewal of human primary erythroid cells. , 2003, Blood.

[45]  C. Stocking,et al.  AML1-ETO Inhibits Maturation of Multiple Lymphohematopoietic Lineages and Induces Myeloblast Transformation in Synergy with ICSBP Deficiency , 2002, The Journal of experimental medicine.

[46]  A. Warren,et al.  Hematopoietic Stem Cell Expansion and Distinct Myeloid Developmental Abnormalities in a Murine Model of the AML1-ETO Translocation , 2002, Molecular and Cellular Biology.

[47]  I. Weissman,et al.  AML1-ETO expression is directly involved in the development of acute myeloid leukemia in the presence of additional mutations , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[48]  T. Yeatman,et al.  Role of Src expression and activation in human cancer , 2000, Oncogene.

[49]  D. Tenen,et al.  Analysis of the role of AML1-ETO in leukemogenesis, using an inducible transgenic mouse model. , 2000, Blood.

[50]  S. Nakamura,et al.  Analysis of genes under the downstream control of the t(8;21) fusion protein AML1-MTG8: overexpression of the TIS11b (ERF-1, cMG1) gene induces myeloid cell proliferation in response to G-CSF. , 2000, Blood.

[51]  J. Downing,et al.  Expression of a knocked-in AML1-ETO leukemia gene inhibits the establishment of normal definitive hematopoiesis and directly generates dysplastic hematopoietic progenitors. , 1998, Blood.

[52]  C. Bloomfield,et al.  Acute myeloid leukemia with 11q23 translocations: myelomonocytic immunophenotype by multiparameter flow cytometry , 1998, Leukemia.

[53]  M. Marín‐Padilla,et al.  Embryonic lethality and impairment of haematopoiesis in mice heterozygous for an AML1-ETO fusion gene , 1997, Nature Genetics.

[54]  A. Hagemeijer,et al.  Acute myeloid leukemia M4 with bone marrow eosinophilia (M4Eo) and inv(16)(p13q22) exhibits a specific immunophenotype with CD2 expression. , 1993, Blood.

[55]  M. Steggerda,et al.  Anthropology and human genetics , 1935 .

[56]  Yiting Lim,et al.  Hedgehog signaling in hematopoiesis. , 2010, Critical reviews in eukaryotic gene expression.

[57]  T. Haferlach,et al.  JAK2 seems to be a typical cooperating mutation in therapy-related t(8;21)/ AML1-ETO-positive AML , 2007, Leukemia.

[58]  Nitin J. Karandikar,et al.  Immunophenotypic identification of acute myeloid leukemia with monocytic differentiation , 2006, Leukemia.

[59]  K. Döhner,et al.  JAK2V617F mutations as cooperative genetic lesions in t(8;21)-positive acute myeloid leukemia. , 2006, Haematologica.

[60]  K. MacKenzie,et al.  The AML1-ETO fusion protein promotes the expansion of human hematopoietic stem cells. , 2002, Blood.