Functional characterization of a novel FGFR1OP‐RET rearrangement in hematopoietic malignancies

The RET (REarranged during Transfection) receptor tyrosine kinase is targeted by oncogenic rearrangements in thyroid and lung adenocarcinoma. Recently, a RET (exon 12) rearrangement with FGFR1OP [fibroblast growth factor receptor 1 (FGFR1) oncogene partner] (exon 12) was identified in one chronic myelomonocytic leukemia (CMML) patient. We report the molecular cloning and functional characterization of a novel FGFR1OP (exon 11)‐RET (exon 11) gene fusion event (named FGFR1OP‐RET), mediated by a reciprocal translocation t(6; 10)(q27; q11), in a patient affected by primary myelofibrosis (PMF) with secondary acute myeloid leukemia (AML). The FGFR1OP‐RET fusion protein displayed constitutive tyrosine kinase and transforming activity in NIH3T3 fibroblasts, and induced IL3‐independent growth and activation of PI3K/STAT signaling in hematopoietic Ba/F3 cells. FGFR1OP‐RET supported cytokine‐independent growth, protection from stress and enhanced self‐renewal of primary murine hematopoietic progenitor and stem cells in vitro. In vivo, FGFR1OP‐RET caused a spectrum of disease phenotypes, with >50% of mice showing a fatal myeloproliferative disorder (MPD). Other phenotypes were leukemia transplantable in secondary recipients, dramatic expansion of the mast cell lineage, and reduction of repopulating activity upon lethal irradiation. In conclusion, FGFR1OP‐RET chimeric oncogenes are endowed with leukemogenic potential and associated to myeloid neoplasms (CMML and PMF/AML).

[1]  J. Issa,et al.  Effects of TET2 mutations on DNA methylation in chronic myelomonocytic leukemia , 2012, Epigenetics.

[2]  P. Campbell,et al.  JAK2 V617F impairs hematopoietic stem cell function in a conditional knock-in mouse model of JAK2 V617F-positive essential thrombocythemia. , 2010, Blood.

[3]  J. Osinga,et al.  RET-familial medullary thyroid carcinoma mutants Y791F and S891A activate a Src/JAK/STAT3 pathway, independent of glial cell line-derived neurotrophic factor. , 2005, Cancer research.

[4]  M. Takahashi,et al.  The GDNF/RET signaling pathway and human diseases. , 2001, Cytokine & growth factor reviews.

[5]  D. Kent,et al.  Mouse models of myeloproliferative neoplasms: JAK of all grades , 2011, Disease Models & Mechanisms.

[6]  F. E. Bertrand,et al.  JAK/STAT, Raf/MEK/ERK, PI3K/Akt and BCR-ABL in cell cycle progression and leukemogenesis , 2004, Leukemia.

[7]  J. Schlessinger.  Cell Signaling by Receptor Tyrosine Kinases , 2000, Cell.

[8]  M. Colombo,et al.  Tumor-intrinsic and -extrinsic roles of c-Kit: mast cells as the primary off-target of tyrosine kinase inhibitors , 2011, Oncogene.

[9]  A. Hagemeijer,et al.  Acquired mutations in TET2 are common in myelodysplastic syndromes , 2009, Nature Genetics.

[10]  Doron Lipson,et al.  Identification of new ALK and RET gene fusions from colorectal and lung cancer biopsies , 2012, Nature Medicine.

[11]  H. Schouten,et al.  Myeloproliferative neoplasia: a review of clinical criteria and treatment. , 2012, The Netherlands journal of medicine.

[12]  M. Santoro,et al.  Drug Insight: small-molecule inhibitors of protein kinases in the treatment of thyroid cancer , 2006, Nature Clinical Practice Endocrinology &Metabolism.

[13]  Yuki Togashi,et al.  RET, ROS1 and ALK fusions in lung cancer , 2012, Nature Medicine.

[14]  I. Matsumura,et al.  Roles for deregulated receptor tyrosine kinases and their downstream signaling molecules in hematologic malignancies , 2008, Cancer science.

[15]  C. Deswarte,et al.  RET fusion genes are associated with chronic myelomonocytic leukemia and enhance monocytic differentiation , 2012, Leukemia.

[16]  F. Lo‐Coco,et al.  retroviral gene transfer into purified murine hematopoietic progenitors PML-RAR induces promyelocytic leukemias with high efficiency following , 2002 .

[17]  H. Hollema,et al.  Ras/ERK1/2-mediated STAT3 Ser727 Phosphorylation by Familial Medullary Thyroid Carcinoma-associated RET Mutants Induces Full Activation of STAT3 and Is Required for c-fos Promoter Activation, Cell Mitogenicity, and Transformation* , 2007, Journal of Biological Chemistry.

[18]  L. Hennighausen,et al.  Essential role for Stat 5 a / b in myeloproliferative neoplasms induced by BCR-ABL 1 and JAK 2 V 617 F in mice , 2012 .

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

[20]  L. Johnson,et al.  The structural basis for control of eukaryotic protein kinases. , 2012, Annual review of biochemistry.

[21]  A. Tefferi.  Myeloproliferative neoplasms 2012: the John M. Bennett 80th birthday anniversary lecture. , 2012, Leukemia research.

[22]  S. Rane,et al.  Hematopoietic cytokine receptor signaling , 2007, Oncogene.

[23]  D. Gilliland,et al.  Detection of mutant TET2 in myeloid malignancies other than myeloproliferative neoplasms: CMML, MDS, MDS/MPN and AML , 2009, Leukemia.

[24]  K. Lennert,et al.  Diagnostic criteria and classification of mastocytosis: a consensus proposal. , 2001, Leukemia research.

[25]  Linzhao Cheng,et al.  Use of green fluorescent protein variants to monitor gene transfer and expression in mammalian cells , 1996, Nature Biotechnology.

[26]  R. Hehlmann,et al.  Treatment of chronic myeloid leukemia in blast crisis , 2008, Haematologica.

[27]  D. Birnbaum,et al.  The t(6;8)(q27;p11) translocation in a stem cell myeloproliferative disorder fuses a novel gene, FOP, to fibroblast growth factor receptor 1. , 1999, Blood.

[28]  K. Nicholas,et al.  Identification and transcript analysis of a novel wallaby (Macropus eugenii) basal-like breast cancer cell line , 2008, Molecular Cancer.

[29]  A. Venditti,et al.  New reciprocal translocation t(6;10) (q27;q11) associated with idiopathic myelofibrosis and eosinophilia. , 2001, Leukemia research.

[30]  C. Eaves,et al.  Insights into the stem cells of chronic myeloid leukemia , 2010, Leukemia.

[31]  R. Mesa,et al.  Transformation of a Chronic Myeloproliferative Neoplasm to Acute Myelogenous Leukemia: Does Anything Work? , 2012, Current Hematologic Malignancy Reports.

[32]  al. et,et al.  Activation of RET as a dominant transforming gene by germline mutations of MEN2A and MEN2B , 1995, Science.

[33]  Yasushi Totoki,et al.  KIF5B-RET fusions in lung adenocarcinoma , 2012, Nature Medicine.

[34]  C. Schumann,et al.  Prognostic significance of ASXL1 mutations in patients with myelodysplastic syndromes. , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[35]  E. Appella,et al.  Impairment of p53 acetylation, stability and function by an oncogenic transcription factor , 2004, The EMBO journal.

[36]  E. Nigg,et al.  A complex of two centrosomal proteins, CAP350 and FOP, cooperates with EB1 in microtubule anchoring. , 2005, Molecular biology of the cell.

[37]  D. Birnbaum,et al.  Myeloproliferative disorder FOP-FGFR1 fusion kinase recruits phosphoinositide-3 kinase and phospholipase Cγ at the centrosome , 2008, Molecular Cancer.

[38]  A. Jankowska,et al.  Mutational spectrum analysis of chronic myelomonocytic leukemia includes genes associated with epigenetic regulation: UTX, EZH2, and DNMT3A. , 2011, Blood.

[39]  J. Jansen,et al.  Mutations in epigenetic regulators in myelodysplastic syndromes , 2012, International Journal of Hematology.

[40]  Li Ding,et al.  Recurrent DNMT3A Mutations in Patients with Myelodysplastic Syndromes , 2011, Leukemia.

[41]  J. Vardiman.  The classification of MDS: from FAB to WHO and beyond. , 2012, Leukemia research.

[42]  L. Hennighausen,et al.  Essential role for Stat5a/b in myeloproliferative neoplasms induced by BCR-ABL1 and JAK2(V617F) in mice. , 2009, Blood.

[43]  C. Bloomfield,et al.  The 2008 revision of the World Health Organization (WHO) classification of myeloid neoplasms and acute leukemia: rationale and important changes. , 2009, Blood.

[44]  A. Pardanani.  Systemic mastocytosis in adults: 2012 Update on diagnosis, risk stratification, and management , 2012, American journal of hematology.

[45]  Stephen L. Abrams,et al.  Roles of the RAF/MEK/ERK and PI3K/PTEN/AKT pathways in malignant transformation and drug resistance. , 2006, Advances in enzyme regulation.

[46]  T. Holak,et al.  Structure of the N-terminal domain of the FOP (FGFR1OP) protein and implications for its dimerization and centrosomal localization. , 2006, Journal of molecular biology.

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

[48]  Matthew B. Wilson,et al.  Control of myeloid differentiation and survival by Stats , 2000, Oncogene.

[49]  M. Santoro,et al.  Molecular characterization of RET/PTC3; a novel rearranged version of the RETproto-oncogene in a human thyroid papillary carcinoma. , 1994, Oncogene.

[50]  L. Lajtha,et al.  Conditions controlling the proliferation of haemopoietic stem cells in vitro , 1977, Journal of cellular physiology.

[51]  D. Birnbaum,et al.  ASXL1 mutation is associated with poor prognosis and acute transformation in chronic myelomonocytic leukaemia , 2010, British journal of haematology.