-
Benjamin H Lee,
Zuzana Tothova,
Ross L Levine,
Kristina Anderson,
Natalija Buza-Vidas,
Dana E Cullen,
Elizabeth P McDowell, Jennifer Adelsperger,
Stefan Fröhling,
Brian J P Huntly,
Miloslav Beran,
Sten Eirik Jacobsen,
D Gary Gilliland
[show abstract]
[hide abstract]
ABSTRACT: Despite their known transforming properties, the effects of leukemogenic FLT3-ITD mutations on hematopoietic stem and multipotent progenitor cells and on hematopoietic differentiation are not well understood. We report a mouse model harboring an ITD in the murine Flt3 locus that develops myeloproliferative disease resembling CMML and further identified FLT3-ITD mutations in a subset of human CMML. These findings correlated with an increase in number, cell cycling, and survival of multipotent stem and progenitor cells in an ITD dose-dependent manner in animals that exhibited alterations within their myeloid progenitor compartments and a block in normal B cell development. This model provides insights into the consequences of constitutive signaling by an oncogenic tyrosine kinase on hematopoietic progenitor quiescence, function, and cell fate.
Cancer Cell 11/2007; 12(4):367-80. · 26.57 Impact Factor
-
David W. Sternberg,
Arghya Ray,
Benjamin H. Lee, Jennifer Adelsperger,
Nicole Duclos,
Elizabeth McDowell,
Naznin Haq,
Patricia Imbach,
Osamu Ohmori,
Toshiyuki Honda,
Markus Warmuth,
Doriano Fabbro,
James D. Griffin,
Johannes Roesel
ASH Annual Meeting 2006; 12/2006
-
Jing Chen,
Benjamin H Lee,
Ifor R Williams,
Jeffery L Kutok,
Constantine S Mitsiades,
Nicole Duclos,
Sarah Cohen, Jennifer Adelsperger,
Rachel Okabe,
Allison Coburn,
Sandra Moore,
Brian J P Huntly,
Doriano Fabbro,
Kenneth C Anderson,
James D Griffin,
Dwight Gary Gilliland
[show abstract]
[hide abstract]
ABSTRACT: Reccurent chromosomal translocation t(4;14) (p16.3;q32.3) occurs in patients with multiple myeloma (MM) and is associated with ectopic overexpression of fibroblast growth factor receptor 3 (FGFR3) that sometimes may contain the activation mutations such as K650E thanatophoric dysplasia type II (TDII). Although there have been significant advances in therapy for MM including the use of proteasome inhibitors, t(4;14) MM has a particularly poor prognosis and most patients still die from complications related to their disease or therapy. One potential therapeutic strategy is to inhibit FGFR3 in those myeloma patients that overexpress the receptor tyrosine kinase due to chromosomal translocation. Here we evaluated PKC412, a small molecule tyrosine kinase inhibitor, for treatment of FGFR3-induced hematopoietic malignancies. PKC412 inhibited kinase activation and proliferation of hematopoietic Ba/F3 cells transformed by FGFR3 TDII or a TEL-FGFR3 fusion. Similar results were obtained in PKC412 inhibition of several different t(4;14)-positive human MM cell lines. Furthermore, treatment with PKC412 resulted in a statistically significant prolongation of survival in murine bone marrow transplant models of FGFR3 TDII-induced pre-B cell lymphoma, or a peripheral T-cell lymphoma associated TEL-FGFR3 fusion-induced myeloproliferative disease. These data indicate that PKC412 may be a useful molecularly targeted therapy for MM associated with overexpression of FGFR3, and perhaps other diseases associated with dysregulation of FGFR3 or related mutants.
Oncogene 01/2006; 24(56):8259-67. · 6.37 Impact Factor
-
Elizabeth H Stover,
Jing Chen,
Benjamin H Lee,
Jan Cools,
Elizabeth McDowell, Jennifer Adelsperger,
Dana Cullen,
Allison Coburn,
Sandra A Moore,
Rachel Okabe,
Doriano Fabbro,
Paul W Manley,
James D Griffin,
D Gary Gilliland
[show abstract]
[hide abstract]
ABSTRACT: AMN107 is a small molecule tyrosine kinase inhibitor developed, in the first instance, as a potent inhibitor of breakpoint cluster region-abelson (BCR-ABL). We tested its effectiveness against fusion tyrosine kinases TEL-platelet-derived growth factor receptorbeta (TEL-PDGFRbeta) and FIP1-like-1 (FIP1L1)-PDGFRalpha, which cause chronic myelomonocytic leukemia and hypereosinophilic syndrome, respectively. In vitro, AMN107 inhibited proliferation of Ba/F3 cells transformed by both TEL-PDGFRbeta and FIP1L1-PDGFRalpha with IC50 (inhibitory concentration 50%) values less than 25 nM and inhibited phosphorylation of the fusion kinases and their downstream signaling targets. The imatinib mesylate-resistant mutant TEL-PDGFRbeta T681I was sensitive to AMN107, whereas the analogous mutation in FIP1L1-PDGFRalpha, T674I, was resistant. In an in vivo bone marrow transplantation assay, AMN107 effectively treated myeloproliferative disease induced by TEL-PDGFRbeta and FIP1L1-PDGFRalpha, significantly increasing survival and disease latency and reducing disease severity as assessed by histopathology and flow cytometry. In summary, AMN107 can inhibit myeloid proliferation driven by TEL-PDGFRbeta and FIP1L1-PDGFRalpha and may be a useful drug for treatment of patients with myeloproliferative disease who harbor these kinase fusions.
Blood 12/2005; 106(9):3206-13. · 9.90 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Constitutively activated forms of the transmembrane receptor tyrosine kinase c-KIT have been associated with systemic mast cell disease, acute myeloid leukemia, and gastrointestinal stromal tumors. Reports of the resistance of the kinase domain mutation D816V to the adenosine triphosphate (ATP)-competitive kinase inhibitor imatinib mesylate prompted us to characterize 14 c-KIT mutations reported in association with human hematologic malignancies for transforming activity in the murine hematopoietic cell line Ba/F3 and for sensitivity to the tyrosine kinase inhibitor PKC412. Ten of 14 c-KIT mutations conferred interleukin 3 (IL-3)-independent growth. c-KIT D816Y and D816V transformed cells were sensitive to PKC412 despite resistance to imatinib mesylate. In these cells, PKC412, but not imatinib mesylate, inhibited autophosphorylation of c-KIT and activation of downstream effectors signal transducer and transcriptional activator 5 (Stat5) and Stat3. Variable sensitivities to PKC412 or imatinib mesylate were observed among other mutants. These findings suggest that PKC412 may be a useful therapeutic agent for c-KIT-positive malignancies harboring the imatinib mesylate-resistant D816V or D816Y activation mutations.
Blood 08/2005; 106(2):721-4. · 9.90 Impact Factor
-
Joseph D Growney,
Hirokazu Shigematsu,
Zhe Li,
Benjamin H Lee, Jennifer Adelsperger,
Rebecca Rowan,
David P Curley,
Jeffery L Kutok,
Koichi Akashi,
Ifor R Williams,
Nancy A Speck,
D Gary Gilliland
[show abstract]
[hide abstract]
ABSTRACT: Homozygous loss of function of Runx1 (Runt-related transcription factor 1 gene) during murine development results in an embryonic lethal phenotype characterized by a complete lack of definitive hematopoiesis. In light of recent reports of disparate requirements for hematopoietic transcription factors during development as opposed to adult hematopoiesis, we used a conditional gene-targeting strategy to effect the loss of Runx1 function in adult mice. In contrast with the critical role of Runx1 during development, Runx1 was not essential for hematopoiesis in the adult hematopoietic compartment, though a number of significant hematopoietic abnormalities were observed. Runx1 excision had lineage-specific effects on B- and T-cell maturation and pronounced inhibition of common lymphocyte progenitor production. Runx1 excision also resulted in inefficient platelet production. Of note, Runx1-deficient mice developed a mild myeloproliferative phenotype characterized by an increase in peripheral blood neutrophils, an increase in myeloid progenitor populations, and extramedullary hematopoiesis composed of maturing myeloid and erythroid elements. These findings indicate that Runx1 deficiency has markedly different consequences during development compared with adult hematopoiesis, and they provide insight into the phenotypic manifestations of Runx1 deficiency in hematopoietic malignancies.
Blood 08/2005; 106(2):494-504. · 9.90 Impact Factor
-
Ross L Levine,
Martha Wadleigh,
Jan Cools,
Benjamin L Ebert,
Gerlinde Wernig,
Brian J P Huntly,
Titus J Boggon,
Iwona Wlodarska,
Jennifer J Clark,
Sandra Moore, [......],
Peter Vandenberghe,
Ruben A Mesa,
Ayalew Tefferi,
James D Griffin,
Michael J Eck,
William R Sellers,
Matthew Meyerson,
Todd R Golub,
Stephanie J Lee,
D Gary Gilliland
[show abstract]
[hide abstract]
ABSTRACT: Polycythemia vera (PV), essential thrombocythemia (ET), and myeloid metaplasia with myelofibrosis (MMM) are clonal disorders arising from hematopoietic progenitors. An internet-based protocol was used to collect clinical information and biological specimens from patients with these diseases. High-throughput DNA resequencing identified a recurrent somatic missense mutation JAK2V617F in granulocyte DNA samples of 121 of 164 PV patients, of which 41 had homozygous and 80 had heterozygous mutations. Molecular and cytogenetic analyses demonstrated that homozygous mutations were due to duplication of the mutant allele. JAK2V617F was also identified in granulocyte DNA samples from 37 of 115 ET and 16 of 46 MMM patients, but was not observed in 269 normal individuals. In vitro analysis demonstrated that JAK2V617F is a constitutively active tyrosine kinase.
Cancer Cell 05/2005; 7(4):387-97. · 26.57 Impact Factor
-
Jing Chen,
Daniel J Deangelo,
Jeffery L Kutok,
Ifor R Williams,
Benjamin H Lee,
Martha Wadleigh,
Nicole Duclos,
Sarah Cohen, Jennifer Adelsperger,
Rachel Okabe, [......],
Pamela S Cohen,
Thomas Meyer,
Doriano Fabbro,
Johannes Roesel,
Lolita Banerji,
James D Griffin,
Sheng Xiao,
Jonathan A Fletcher,
Richard M Stone,
D Gary Gilliland
[show abstract]
[hide abstract]
ABSTRACT: Human stem cell leukemia-lymphoma syndrome usually presents itself as a myeloproliferative disorder (MPD) that evolves to acute myeloid leukemia and/or lymphoma. The syndrome associated with t(8;13)(p11;q12) results in expression of the ZNF198-fibroblast growth factor receptor (FGFR) 1 fusion tyrosine kinase. Current empirically derived cytotoxic chemotherapy is inadequate for treatment of this disease. We hypothesized that small-molecule inhibitors of the ZNF198-FGFR1 fusion would have therapeutic efficacy. We characterized the transforming activity of ZNF198-FGFR1 in hematopoietic cells in vitro and in vivo. Expression of ZNF198-FGFR1 in primary murine hematopoietic cells caused a myeloproliferative syndrome in mice that recapitulated the human MPD phenotype. Transformation in these assays, and activation of the downstream effector molecules PLC-gamma, STAT5, and phosphatidylinositol 3-kinase/AKT, required the proline-rich domains, but not the ZNF domains, of ZNF198. A small-molecule tyrosine kinase inhibitor, PKC412 (N-benzoyl-staurosporine) effectively inhibited ZNF198-FGFR1 tyrosine kinase activity and activation of downstream effector pathways, and inhibited proliferation of ZNF198-FGFR1 transformed Ba/F3 cells. Furthermore, treatment with PKC412 resulted in statistically significant prolongation of survival in the murine model of ZNF198-FGFR1-induced MPD. Based in part on these data, PKC412 was administered to a patient with t(8;13)(p11;q12) and was efficacious in treatment of progressive myeloproliferative disorder with organomegaly. Therefore, PKC412 may be a useful therapy for treatment of human stem cell leukemia-lymphoma syndrome.
Proceedings of the National Academy of Sciences 11/2004; 101(40):14479-84. · 9.68 Impact Factor
