MOLECULAR PATHOGENESIS & THERAPY OF JAK2 V617F-INDUCED MYELOPROLIFERATIVE DISEASE
Richard A Van etten, Chief, Hematology/oncology
Tufts Medical Center, 800 Washington St, Boston, Ma 02111-1526
Grant 5R01HL089747-03 from National Heart, Lung, And Blood Institute
Abstract: Chronic myeloid leukemia (CML) is a myeloproliferative disease (MPD) associated with BCR-ABL, an activated tyrosine kinase that is the product of the Philadelphia chromosome. A mouse model of CML, where BCR-ABL is expressed by retroviral bone marrow transduction and transplantation, demonstrated that BCR-ABL is the direct cause of CML, and motivated the development of imatinib, an ABL kinase inhibitor that has revolutionized the treatment of CML patients. Polycythemia vera (PV), essential thromobocythemia (ET), and chronic idiopathic myelofibrosis (CIMF) are MPDs that are as prevalent as CML, but their pathogenesis is unclear, and their clinical course is complicated by bleeding, thrombosis, and progression to acute leukemia or bone marrow failure. Current therapy for these MPDs is also inadequate, ranging from phlebotomy and myelosuppressive therapy for PV and ET, to transfusions and supportive care for CIMF. Recently, a somatic activating mutation (V617F) in the JAK2 tyrosine kinase was discovered in most patients with PV and some patients with ET and CIMF. However, the role of JAK2 V617F in the pathogenesis and potential therapy of these diseases is not known. A mouse retroviral transduction/transplantation model of MPD induced by JAK2 V617F has recently been developed by our laboratory. Using the model, it has been demonstrated that JAK2 V617F recapitulates the entire spectrum of erythroid abnormalities of PV in mice, implicating it as the direct and principal cause of human PV. In this application, this model system will be used to address some fundamental questions about the molecular pathogenesis and therapy of JAK2 V617F-induced MPD that would be difficult or impossible to approach through human studies. Specifically, the application proposes to (1) use a genetic approach to determine the pathways downstream of JAK2 V617F that are required for MPD, with the ultimate goal of validating additional therapeutic targets; and (2) test the therapeutic activity of molecularly targeted drugs in JAK2 V617F-induced MPD, including candidate JAK2 inhibitors. These experiments will yield new knowledge about the biology and treatment of this important group of MPDs, and should provide important information to guide the clinical development of JAK2 kinase inhibitors and other molecularly targeted therapies for MPD. Polycythemia vera (PV) is a human disease where too many red blood cells are produced. Current therapy for PV is inadequate due to blood clotting problems and development of leukemia. In this proposal, a mouse model of PV will be used to better understand the basic cause of PV, and to test new treatments for the disease, which could lead to better treatments for PV patients
Keywords: A Mouse; ATP[{..}]protein-tyrosine O-phosphotransferase; Acute leukemia; Address; Agnogenic Myeloid Metaplasia; Anemia; Antibodies; Assay; BCR-ABL; BCR-ABL Kinase; BCR-ABL Oncoprotein; BCR-ABL Protein Tyrosine Kinase; BCR/ABL; Bcr-Abl tyrosine kinase; Bioassay; Biologic Assays; Biological Assay; Biological Models; Biology; Bizzozero`s corpuscle/cell; Bleeding; Bleeding Time; Bleeding time procedure; Blood (Leukemia); Blood Clot; Blood Clotting; Blood Coagulation Disorders; Blood Neutrophil; Blood Plasma; Blood Platelet Count; Blood Platelet Number; Blood Platelets; Blood Polymorphonuclear Neutrophil; Blood Segmented Neutrophil; Blood coagulation; Blood erythrocyte; Blood leukocyte; Blood megakaryocyte; Blood normocyte; Body Tissues; Bone Marrow; Bone marrow failure; CYTOGEN; Cells; Cessation of life; Chronic Idiopathic Myelofibrosis; Chronic Myeloid Leukemia; Clinical; Clotting; Coagulation; Coagulation Disorder; Coagulation Process; Coagulopathy; Commit; Cytogenetic; Cytogenetics; Death; Deetjeen`s body; Development; Disease; Disease remission; Disorder; Drug Delivery; Drug Delivery Systems; Drug Targeting; Drug Targetings; Dysfunction; EPH- and ELK-Related Tyrosine Kinase; EPH-and ELK-Related Kinase; EPHA8; EPO-R; EphA8 Protein; Ephrin Type-A Receptor 8; Ephrin Type-A Receptor 8 Precursor; Erythremia; Erythrocytes; Erythrocytic; Erythroid; Erythropoietin Receptor; Essential Thrombocytemia; Essential Thrombocytosis; Event; Fibrosis, Bone Marrow; Functional disorder; Fusion Proteins, bcr-abl; Genetic; Genetic Alteration; Genetic Change; Genetic defect; Goals; HEK3; Hayem`s elementary corpuscle; Hemorrhage; Hemorrhagic Thrombocythemia; Hemostasis; Hemostatic Agents; Hemostatic function; Hemostatics; Heterophil Granulocyte; Human; Human, General; Idiopathic Bone Marrow Fibrosis; Idiopathic Myelofibrosis; Imatinib; Inbred Strains Mice; JAK2; JAK2 gene; Knowledge; Laboratories; Lead; Leukemia, Granulocytic, Chronic; Leukemias, General; Leukocytes; Leukocytosis; Mammals, Mice; Man (Taxonomy); Man, Modern; Marrow Neutrophil; Marrow erythrocyte; Marrow leukocyte; Marrow platelet; Megakaryocytes; Megalokaryocyte; Mice; Model System; Modeling; Models, Biologic; Molecular; Morbidity; Morbidity - disease rate; Mother Cells; Murine; Mus; Mutation; Myelocytic Leukemia, Chronic; Myelofibrosis; Myelogenous Leukemia, Chronic; Myeloid Disease; Myeloid Leukemia, Chronic; Myeloid Malignancy; Myeloid Neoplasm; Myeloid Tumor; Myeloproliferative Disorders; Myeloproliferative Tumors; Myeloproliferative disease; Myelosclerosis; Myelosclerosis with Myeloid Metaplasia; Myelosuppressive Therapy; Neutrophilia; Neutrophilic Granulocyte; Neutrophilic Leukocyte; Nonleukemic Myelosis; Osler-Vaquez Disease; PTK; PTK Receptors; Pancytopenia; Pathogenesis; Pathway interactions; Patients; Pb element; Ph 1 Chromosome; Ph1 Chromosome; Phase; Philadelphia Chromosome; Phlebotomy; Physiopathology; Plasma; Platelet Count; Platelet Count measurement; Platelet Number; Platelets; Polycythemia; Polycythemia Rubra Vera; Polycythemia Vera; Polymorph; Polymorphonuclear Cell; Polymorphonuclear Leukocytes; Polymorphonuclear Neutrophils; Primary Myelofibrosis; Primary Thrombocytosis; Progenitor Cells; Protein Tyrosine Kinase; Protein Tyrosine Kinase EEK; Protein-Tyrosine Kinases, src; RMSN; RTK; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Red Blood Cells; Red Cell; Red Cell Mass result; Red Cell Masses; Red blood corpuscule; Red cell of marrow; Remission; Reticulocytosis; Reticuloendothelial System, Bone Marrow; Reticuloendothelial System, Erythrocytes; Reticuloendothelial System, Leukocytes; Reticuloendothelial System, Platelets; Reticuloendothelial System, Serum, Plasma; Role; Serum, Plasma; Signal Pathway; Signaling Molecule; Somatic Mutation; Stem cells; Supportive Therapy; Supportive care; TRNSF; Tail; Testing; Therapeutic; Therapeutic Effect; Thrombocytes; Thrombocythemia, Essential; Thrombocythemia, Hemorrhagic; Thrombocythemia, Idiopathic; Thrombocythemia, Primary; Thrombosis; Tissues; Transfusion; Transmembrane Receptor Protein Tyrosine Kinase; Transplantation; Tyrosine Kinase; Tyrosine Kinase Growth Factor Receptor; Tyrosine Kinase Linked Receptors; Tyrosine Kinase Receptors; Tyrosine-Protein Kinase Receptor EEK; Tyrosine-Specific Protein Kinase; Tyrosylprotein Kinase; Venous blood sampling; White Blood Cells; White Cell; base; bcr-abl Fusion Proteins; blood corpuscles; blood loss; clotting disorder; disease/disorder; experiment; experimental research; experimental study; genome mutation; heavy metal Pb; heavy metal lead; human disease; hydroxyaryl protein kinase; in vivo; inhibitor; inhibitor/antagonist; kinase inhibitor; leukemia; mouse model; myeloproliferative neoplasm; myelosis non-leukemic; neutrophil; pathophysiology; pathway; progenitor; research study; retroviral transduction; self-renewal; social role; src Kinases; src Tyrosine Kinases; src-Family Kinases; src-Family Tyrosine Kinases; therapeutic target; thrombocyte/platelet; transplant; tyrosyl protein kinase; white blood cell; white blood corpuscle
Project start date: 2008-04-01
Project end date: 2013-03-31
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2011
PFA/PA: PA-07-070
5R01HL089747-03 (2010): $402500
Sponsored Links Excellgen http://Excellgen.com
MOLECULAR PATHOGENESIS & THERAPY OF JAK2 V617F-INDUCED MYELOPROLIFERATIVE DISEASE
Richard A Van etten, Chief, Hematology/oncology
Tufts Medical Center, 800 Washington St, Boston, Ma 02111-1526
Grant 5R01HL089747-02 from National Heart, Lung, And Blood Institute
Abstract: Chronic myeloid leukemia (CML) is a myeloproliferative disease (MPD) associated with BCR-ABL, an activated tyrosine kinase that is the product of the Philadelphia chromosome. A mouse model of CML, where BCR-ABL is expressed by retroviral bone marrow transduction and transplantation, demonstrated that BCR-ABL is the direct cause of CML, and motivated the development of imatinib, an ABL kinase inhibitor that has revolutionized the treatment of CML patients. Polycythemia vera (PV), essential thromobocythemia (ET), and chronic idiopathic myelofibrosis (CIMF) are MPDs that are as prevalent as CML, but their pathogenesis is unclear, and their clinical course is complicated by bleeding, thrombosis, and progression to acute leukemia or bone marrow failure. Current therapy for these MPDs is also inadequate, ranging from phlebotomy and myelosuppressive therapy for PV and ET, to transfusions and supportive care for CIMF. Recently, a somatic activating mutation (V617F) in the JAK2 tyrosine kinase was discovered in most patients with PV and some patients with ET and CIMF. However, the role of JAK2 V617F in the pathogenesis and potential therapy of these diseases is not known. A mouse retroviral transduction/transplantation model of MPD induced by JAK2 V617F has recently been developed by our laboratory. Using the model, it has been demonstrated that JAK2 V617F recapitulates the entire spectrum of erythroid abnormalities of PV in mice, implicating it as the direct and principal cause of human PV. In this application, this model system will be used to address some fundamental questions about the molecular pathogenesis and therapy of JAK2 V617F-induced MPD that would be difficult or impossible to approach through human studies. Specifically, the application proposes to (1) use a genetic approach to determine the pathways downstream of JAK2 V617F that are required for MPD, with the ultimate goal of validating additional therapeutic targets; and (2) test the therapeutic activity of molecularly targeted drugs in JAK2 V617F-induced MPD, including candidate JAK2 inhibitors. These experiments will yield new knowledge about the biology and treatment of this important group of MPDs, and should provide important information to guide the clinical development of JAK2 kinase inhibitors and other molecularly targeted therapies for MPD. Polycythemia vera (PV) is a human disease where too many red blood cells are produced. Current therapy for PV is inadequate due to blood clotting problems and development of leukemia. In this proposal, a mouse model of PV will be used to better understand the basic cause of PV, and to test new treatments for the disease, which could lead to better treatments for PV patients
Keywords: A Mouse; ATP[{..}]protein-tyrosine O-phosphotransferase; Acute leukemia; Address; Agnogenic Myeloid Metaplasia; Anemia; Antibodies; Assay; BCR-ABL; BCR-ABL Kinase; BCR-ABL Oncoprotein; BCR-ABL Protein Tyrosine Kinase; BCR/ABL; Bcr-Abl tyrosine kinase; Bioassay; Biologic Assays; Biological Assay; Biological Models; Biology; Bizzozero`s corpuscle/cell; Bleeding; Bleeding Time; Bleeding time procedure; Blood (Leukemia); Blood Clot; Blood Clotting; Blood Coagulation Disorders; Blood Neutrophil; Blood Plasma; Blood Platelet Count; Blood Platelet Number; Blood Platelets; Blood Polymorphonuclear Neutrophil; Blood Segmented Neutrophil; Blood coagulation; Blood erythrocyte; Blood leukocyte; Blood megakaryocyte; Blood normocyte; Body Tissues; Bone Marrow; Bone marrow failure; CYTOGEN; Cells; Cessation of life; Chronic Idiopathic Myelofibrosis; Chronic Myeloid Leukemia; Clinical; Clotting; Coagulation; Coagulation Disorder; Coagulation Process; Coagulopathy; Commit; Cytogenetic; Cytogenetics; Death; Deetjeen`s body; Development; Disease; Disease remission; Disorder; Drug Delivery; Drug Delivery Systems; Drug Targeting; Drug Targetings; Dysfunction; EPH- and ELK-Related Tyrosine Kinase; EPH-and ELK-Related Kinase; EPHA8; EPO-R; EphA8 Protein; Ephrin Type-A Receptor 8; Ephrin Type-A Receptor 8 Precursor; Erythremia; Erythrocytes; Erythrocytic; Erythroid; Erythropoietin Receptor; Essential Thrombocytemia; Essential Thrombocytosis; Event; Fibrosis, Bone Marrow; Functional disorder; Fusion Proteins, bcr-abl; Genetic; Genetic Alteration; Genetic Change; Genetic defect; Goals; HEK3; Hayem`s elementary corpuscle; Hemorrhage; Hemorrhagic Thrombocythemia; Hemostasis; Hemostatic Agents; Hemostatic function; Hemostatics; Heterophil Granulocyte; Human; Human, General; Idiopathic Bone Marrow Fibrosis; Idiopathic Myelofibrosis; Imatinib; Inbred Strains Mice; JAK2; JAK2 gene; Knowledge; Laboratories; Lead; Leukemia, Granulocytic, Chronic; Leukemias, General; Leukocytes; Leukocytosis; Mammals, Mice; Man (Taxonomy); Man, Modern; Marrow Neutrophil; Marrow erythrocyte; Marrow leukocyte; Marrow platelet; Megakaryocytes; Megalokaryocyte; Mice; Mice, Inbred Strains; Model System; Modeling; Models, Biologic; Molecular; Morbidity; Morbidity - disease rate; Mother Cells; Murine; Mus; Mutation; Myelocytic Leukemia, Chronic; Myelofibrosis; Myelogenous Leukemia, Chronic; Myeloid Disease; Myeloid Leukemia, Chronic; Myeloid Malignancy; Myeloid Neoplasm; Myeloid Tumor; Myeloproliferative Disorders; Myeloproliferative Tumors; Myeloproliferative disease; Myelosclerosis; Myelosclerosis with Myeloid Metaplasia; Myelosuppressive Therapy; Neutrophilia; Neutrophilic Granulocyte; Neutrophilic Leukocyte; Nonleukemic Myelosis; Osler-Vaquez Disease; PTK; PTK Receptors; Pancytopenia; Pathogenesis; Pathway interactions; Patients; Pb element; Ph 1 Chromosome; Ph1 Chromosome; Phase; Philadelphia Chromosome; Phlebotomy; Physiopathology; Plasma; Platelet Count; Platelet Count measurement; Platelet Number; Platelets; Polycythemia; Polycythemia Rubra Vera; Polycythemia Vera; Polymorph; Polymorphonuclear Cell; Polymorphonuclear Leukocytes; Polymorphonuclear Neutrophils; Primary Myelofibrosis; Primary Thrombocytosis; Progenitor Cells; Protein Tyrosine Kinase; Protein Tyrosine Kinase EEK; Protein-Tyrosine Kinases, src; RMSN; RTK; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Red Blood Cells; Red Cell; Red Cell Mass result; Red Cell Masses; Red blood corpuscule; Red cell of marrow; Remission; Reticulocytosis; Reticuloendothelial System, Bone Marrow; Reticuloendothelial System, Erythrocytes; Reticuloendothelial System, Leukocytes; Reticuloendothelial System, Platelets; Reticuloendothelial System, Serum, Plasma; Role; Serum, Plasma; Signal Pathway; Signaling Molecule; Somatic Mutation; Stem cells; Supportive Therapy; Supportive care; TRNSF; Tail; Testing; Therapeutic; Therapeutic Effect; Thrombocytes; Thrombocythemia, Essential; Thrombocythemia, Hemorrhagic; Thrombocythemia, Idiopathic; Thrombocythemia, Primary; Thrombosis; Tissues; Transfusion; Transmembrane Receptor Protein Tyrosine Kinase; Transplantation; Tyrosine Kinase; Tyrosine Kinase Growth Factor Receptor; Tyrosine Kinase Linked Receptors; Tyrosine Kinase Receptors; Tyrosine-Protein Kinase Receptor EEK; Tyrosine-Specific Protein Kinase; Tyrosylprotein Kinase; Venous blood sampling; White Blood Cells; White Cell; base; bcr-abl Fusion Proteins; blood corpuscles; blood loss; clotting disorder; disease/disorder; experiment; experimental research; experimental study; genome mutation; heavy metal Pb; heavy metal lead; human disease; hydroxyaryl protein kinase; in vivo; inhibitor; inhibitor/antagonist; kinase inhibitor; leukemia; mouse model; myeloproliferative neoplasm; myelosis non-leukemic; neutrophil; pathophysiology; pathway; progenitor; research study; retroviral transduction; self-renewal; social role; src Kinases; src Tyrosine Kinases; src-Family Kinases; src-Family Tyrosine Kinases; therapeutic target; thrombocyte/platelet; transplant; tyrosyl protein kinase; white blood cell; white blood corpuscle
Project start date: 2008-04-01
Project end date: 2013-03-31
Budget start date: 1-APR-2009
Budget end date: 31-MAR-2010
PFA/PA: PA-07-070
5R01HL089747-02 (2009): $402500
Grants awarded to Richard A Van etten
Richard A Van etten
Tufts Medical Center, 800 Washington St, Boston, Ma 02111-1526
Grant 5T32CA009429-27 from National Cancer Institute
Abstract: This is a revised application from the William Dameshek Division of Hematology/Oncology at Tufts Medical Center for competitive renewal of our long-standing training grant, T32 CA09429, entitled "Research Training in Oncology". The long-term objective of the application has not changed, which is to continue the training and mentoring of physicians in the Hematology/Oncology Division Fellowship Program in basic and translational cancer science, in order to prepare them for independent careers in academic hematology/oncology. The program is based in the Division of Hematology/Oncology at Tufts Medical Center (Tufts MC), the principal teaching hospital for Tufts University School of Medicine (TUSM). The training program is a highly structured research experience that is centered around laboratory investigation within the research group of a faculty mentor. The laboratory component is complimented by formal coursework, other didactic training, and participation in seminars, workshops, and meetings. The progress of each trainee is closely monitored by a research committee. The Program currently has 26 research faculty mentors drawn from members of the Division and from other clinical and basic science departments at Tufts MC and TUSM, a significant increase from 19 Program faculty in the last competitive renewal in 2002. This reflects a new emphasis on cancer in the strategic plans of Tufts MC and TUSM, leading to the establishment of the Molecular Oncology Research Institute, and a major institutional effort to obtain National Cancer Institute Comprehensive Cancer Center designation for the Tufts Medical Center Cancer Center. The Program faculty mentors represent a major strength of the application, and are carefully selected on the basis of their training record and for their specific research interests in neoplasia, which span the spectrum from oncogenes and tumor suppressors, signal transduction, cell cycle regulation and apoptosis, angiogenesis and metastasis, animal models of hematologic malignancies and solid tumors, and preclinical testing of molecularly targeted cancer therapeutics. This training Program has been highly successful in preparing our fellows for careers in academic oncology. Former trainees now head major departments in academia and in NCI designated cancer centers, are on faculty at academic institutions in the US and elsewhere, and hold senior management positions in the oncology pharmaceutical field. RELEVANCE The treatment of cancer has been revolutionized over the past ten years through the development of drugs that specifically target cancer cells. The United States has a great need for cancer doctors who work at the interface between cancer patients and experimental treatments, and who can discover and apply the next generation of therapies. This grant application will support the training of such physicians at Tufts Medical Center, continuing a program that has generated leaders in this field for more than a quarter century
Keywords: Cancer, Oncology; Research Training; oncology
Project start date: 1981-09-01
Project end date: 2014-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: PA-08-226
5T32CA009429-27 (2010): $237148
3T32CA009429-27S1 (2010): $69319
LEUKEMOGENESIS BY BCR/ABL GENES
Richard A Van etten, Professor - Dept Of Medicine/hematology-
Immune Disease Institute, Inc. 800 Huntington Ave Boston, Ma 02115
Grant 5R01CA057593-03 from National Cancer Institute IRG: HEM
Abstract: The Philadelphia chromosome-positive human leukemias, including the myeloproliferative syndrome chronic myelogenous leukemia (CML), and acute lymphoid and myeloid leukemias, constitute an important group of hematologic neoplasms whose clinical behavior has been very well characterized. As the product of the Philadelphia chromosome, activated bcr/abl fusion genes have been implicated in the pathogenesis of the Philadelphia-positive leukemias, but the way in which these abnormal genes induce leukemia is unknown. The long-term objective of this proposal is to gain an understanding of the cellular and molecular mechanism of leukemogenesis by bcr/abl genes by expression of these genes in the hematopoietic system of mice. The two major forms of bcr/abl will be expressed in the murine hematopoietic system by the use of retroviral gene transfer and transgenic mice. In the first Specific Aim, the target cells for the induction of specific hematological malignancies in mice by the P210(bcr/abl) gene will be defined. The multipotential cell which appears to be the target for the induction of a CML-like syndrome in mice by the P210(bcr/abl) gene will be characterized by lineage analysis in primary animals, by direct purification of target cells, and by the complementary method of expression of bcr/abl in transgenic mice. In the second Aim, the difference in disease spectrum induced by the P210(bcr/abl) gene and the P190(bcr/abl) gene will be carefully defined using a similar strategy. The determinants in bcr which allow the induction of a myeloproliferative syndrome as opposed to acute leukemia will be identified by a deletion mapping approach. These experiments, which are difficult or impossible to carry out on humans, should clarify the mechanism of leukemogenesis by bcr/abl genes, offering insight into the pathogenesis of the human Philadelphia-positive leukemias and suggesting new therapeutic approaches to these diseases.
Keywords: carcinogenesis, chronic myelogenous leukemia, myeloproliferative neoplasm, neoplasm /cancer genetics, neoplastic transformation, oncogene, disease model, fusion gene, gene expression, genetic promoter element, hematopoietic stem cell, protein tyrosine kinase, bone marrow, flow cytometry, gene complementation, genetic mapping, in situ hybridization, laboratory mouse, polymerase chain reaction, protein purification, southern blotting, tissue /cell culture, transfection, transgenic animal
Project start date: 1995-07-01
Project end date: 1998-06-30
5R01CA057593-03 (1997): $301106
NOVEL CELLULAR THERAPIES FOR PH+ LEUKEMIA
Richard A Van etten
Tufts Medical Center, 800 Washington St, Boston, Ma 02111-1526
Grant 5R01HL093981-02 from National Heart, Lung, And Blood Institute
Abstract: The Philadelphia chromosome-positive (Ph+) leukemias, including chronic myeloid leukemia (CML) and Ph+ B- cell acute lymphoblastic leukemia (B-ALL), are prevalent blood cancers for which our current therapies are inadequate. While BCR-ABL tyrosine kinase inhibitors (TKIs) such as imatinib mesylate have replaced hematopoietic stem cell transplantation (HSCT) as initial therapy for CML, complete molecular remissions are rare and acquired resistance to TKI therapy is a significant clinical problem. Eligible Ph+ B-ALL patients undergo allogeneic HSCT in first remission following chemotherapy, but over half will relapse. Hence, it is likely that current therapy will not cure most Ph+ leukemia patients, and effective methods to eradicate residual leukemia are needed. This is a collaborative, dual-PI application from two senior clinician-scientists focused on the preclinical development of novel transfusion-based cellular therapies designed to eliminate residual disease in Ph+ leukemia patients, leading to permanent cure. To accomplish these goals, we will utilize a well- characterized mouse model of CML and Ph+ B-ALL to determine the efficacy and cellular mechanisms of adoptive immunotherapy with allogeneic T-lymphocytes or natural killer (NK) cells. To extend the benefits of NK cell immunotherapy to lymphoid malignancies, we will continue the preclinical development of NK cells engineered to express activating receptors that recognize ligands on B-lymphoid leukemia cells (CD19 and CD20), and test their efficacy in an in vivo immunotherapy model against human Ph+ B-ALL in NOD/SCID/¿c mice. Finally, having demonstrated that HSC in CML are uniquely dependent on selectins and their ligands for homing and engraftment in mice, we will develop clinically relevant methods for blocking engraftment of leukemic stem cells without affecting normal HSC. Throughout the application, a major emphasis will be placed on translation of novel findings to the clinic. Together, these studies should yield important new knowledge that will improve the effectiveness of adoptive immunotherapy and autologous HSCT for Ph+ leukemia, and increase the proportion of patients that are cured of their disease. PHS 398/2590 (Rev. 11/07) Page 2 Continuation Format Page Program Director/Principal Investigator (Last, First, Middle) Van Etten R.A./Klingemann H.K
Keywords: ABL Tyrosine Kinase; ABL1; ATGN; Adoptive Cellular Immunotherapy; Adoptive Immunotherapy; Affect; Allogenic; Allografting; Antibody Fragments; Antigen Receptors; Antigens; Autologous; B Cell Antigen CD19; B-ALL; B-Cell Acute Lymphoblastic Leukemia; B-Cell Lymphoblastic Leukemia; B-Cell Precursor Type Acute Leukemia; B-Cell Type Acute Leukemia; B-Lymphocyte Antigen CD19; B-Lymphocyte Surface Antigen B4; B-Lymphocytic Leukemia, Acute; BCR-ABL; BCR-ABL Oncoprotein; BCR-ABL Protein Tyrosine Kinase; BCR/ABL; Binding; Binding (Molecular Function); Blood (Leukemia); Blood Cells; Blood Precursor Cell; Blood, Cord; Bone Marrow; Bp35; Burkitt Leukemia; Burkitt`s Cell Leukemia; CD19; CD19 Antigens; CD19 gene; CD19 molecule; CD20; CD3; CD3 Antigens; CD3 Complex; CD3 molecule; CD44; CD44 gene; Categories; Cells; Chimera; Chimera organism; Chromosomes; Chronic Myeloid Leukemia; Clinic; Clinical; Cytotoxic cell; Development; Differentiation Antigen CD19; Disease; Disease remission; Disorder; Drug Delivery; Drug Delivery Systems; Drug Targeting; Drug Targetings; Drugs; Dysmyelopoietic Syndromes; Effectiveness; Engraftment; FAB L3; FLR; Failure (biologic function); Fusion Proteins, bcr-abl; Goals; Grant; HSC transplantation; Hematopoietic Cell Tumor; Hematopoietic Malignancies; Hematopoietic Neoplasms; Hematopoietic Neoplasms including Lymphomas; Hematopoietic Stem Cell Transplantation; Hematopoietic Tumor; Hematopoietic and Lymphoid Cell Neoplasm; Hematopoietic and Lymphoid Neoplasms; Hematopoietic stem cells; Hematopoietic, Including Myeloma; Homing; Human; Human, General; ITX; Imatinib mesylate; Immune system; Immunoglobulin Fragments; Immunologically Directed Therapy; Immunotherapy; Immunotherapy, Adoptive; K lymphocyte; Killings; Knowledge; L3 Acute Lymphoblastic Leukemia; L3 Acute Lymphocytic Leukemia; L3 Acute Lymphogenous Leukemia; L3 Acute Lymphoid Leukemia; Leu 12; Leu-16; Leukemia, B-Cell, Acute; Leukemia, Granulocytic; Leukemia, Granulocytic, Chronic; Leukemia, Lymphoblastic, Burkitt-Type; Leukemia, Lymphocytic; Leukemias, General; Life; Ligands; Lymphoblastic Leukemia; Lymphocytic Leukemia, B-Cell, Acute; Lymphocytic Leukemia, L3; Lymphoid; Lymphoid Cell; Lymphoid Leukemia; MDU3; MS4A1; MS4A1 gene; MS4A2; Malignant; Malignant - descriptor; Malignant Cell; Malignant Hematopoietic Neoplasm; Malignant lymphoid neoplasm; Mammals, Mice; Man (Taxonomy); Man, Modern; Mediating; Medication; Methods; Mice; Minor Histocompatibility Antigens; Minor Histocompatibility Peptides; Modeling; Molecular; Molecular Interaction; Mother Cells; Murine; Mus; Myelocytic Leukemia; Myelocytic Leukemia, Chronic; Myelodysplastic Syndromes; Myelogenous Leukemia; Myelogenous Leukemia, Chronic; Myeloid Disease; Myeloid Leukemia; Myeloid Leukemia, Chronic; Myeloid Malignancy; Myeloid Neoplasm; Myeloid Tumor; Myeloproliferative Disorders; Myeloproliferative Tumors; Myeloproliferative disease; NK Cells; NOD/SCID mouse; Natural Killer Cells; Nature; Non-Lymphoblastic Leukemia; Non-Lymphocytic Leukemia; OKT3 antigen; P150; PTK Inhibitors; Patients; Peripheral Blood Cell; Pgp1; Ph 1 Chromosome; Ph1 Chromosome; Pharmaceutic Preparations; Pharmaceutical Preparations; Philadelphia Chromosome; Precursor B Lymphoblastic Leukemia; Principal Investigator; Procedures; Progenitor Cells; Progenitor Cells, Hematopoietic; Protein Tyrosine Kinase Inhibitors; Proto-Oncogene Tyrosine -Protein Kinase ABL; Protocol; Protocols documentation; Protocols, Treatment; RGM; RMSN; Receptor Protein; Regimen; Relapse; Remission; Residual; Residual Tumors; Residual state; Resistance; Reticuloendothelial System, Bone Marrow; Scientist; Selectins; Series; Smoldering Leukemia; Source; Stem cells; T-Cells; T-Lymphocyte; T3 Antigens; T3 Complex; T3 molecule; TK Inhibitors; TRNSF; Testing; Thymus-Dependent Lymphocytes; Transfusion; Translations; Transplantation; Treatment Protocols; Treatment Regimen; Treatment Schedule; Tumors, Residual; Tyrosine Kinase Inhibitor; Umbilical Cord Blood; adoptive cell immunotherapy; base; bcr-abl Fusion Proteins; blood cancer; body system, allergic/immunologic; c-ABL Protein; cancer cell; cell engineering; cellular engineering; chemotherapy; clinical relevance; clinically relevant; cytotoxic; design; designing; disease/disorder; drug/agent; efficacy testing; failure; fetal cord blood; high risk; immune therapy; immunogen; improved; in vivo; intervention design; leukemia; lymphatic leukemia; lymphogenous leukemia; lymphoid malignancy; mouse model; myelodysplasia; myeloid granulocytic leukemia; myeloproliferative neoplasm; myelosis; novel; organ system, allergic/immunologic; pre-clinical; preclinical; public health relevance; reagent testing; receptor; residual disease; resistant; therapy design; thymus derived lymphocyte; transplant; treatment design; tyrosine kinase ABL1
Relevance: Despite the advent of new "targeted" drugs for the treatment of blood cancer (leukemia), many patients will fail therapy and relapse. The goals of this proposal are to develop and test new leukemia therapies, based on transfusion of cells from the blood and immune system, which are designed to eliminate small numbers of cancer cells that persist in drug-treated patients. These "cellular therapies" should provide important new treatment choices that increase the chance of permanent cure for people living with leukemia
Project start date: 2009-09-15
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: PA-07-070
5R01HL093981-02 (2010): $724784
1R01HL093981-01A1 (2009): $707089
STUDIES OF BCR/ABL LEUKEMOGENESIS IN MICE
Richard A Van etten, Professor - Dept Of Medicine/hematology-
Immune Disease Institute, Inc.
800 Huntington Ave
boston, Ma 02115
Grant 1R01CA090576-01 from National Cancer Institute IRG: PTHB
Abstract: The human Philadelphia (Ph) chromosome-positive leukemias, including chronic myeloid leukemia (CML) and B-cell acute lymphoblastic leukemia, are among the most common hematological malignancies, and current therapy for these diseases is inadequate. Expression of the product of the t(9;22) Ph chromosome, the BCR!ABL fusion gene, in the hematopoietic system of mice by generation of transgenic mice or through retroviral transduction and transplantation of bone marrow has demonstrated that BCR/ABL is a leukemia specific oncogene and the direct cause of CML. The long term objective of this application is a more complete molecular and genetic understanding of the pathophysiology of human Ph-positive leukemias, particularly the myeloproliferative disease CML. These goals will be accomplished by the use of a retroviral bone marrow infection/transplantation mouse model system that accurately and quantitatively models both human CML and Ph-positive B-lymphoid leukemia, and will have two Specific Aims. In the first Aim, the signaling pathways important for leukemogenesis by BCR/ABL will be identified by testing BCR/ABL mutants, by further analysis of the requirement for direct binding of the Grb2 adapter protein by the Bcr/Abl fusion protein, and through the use of mice with germline mutations in signaling molecules. In the second Aim, the bone marrow target cells that initiate the CML-like disease and B-lymphoid leukemia induced by BCR/ABL will be characterized and isolated by physical and immunological methods. These investigations will add important new information to our understanding of these leukemias, that would be difficult if not impossible to obtain from studies in vitro, in cultured cells, or in primary human CML cells. This knowledge will be valuable for improving the diagnosis and treatment of the Ph-positive leukemias
Keywords: acute lymphocytic leukemia, chromosome translocation, chronic myelogenous leukemia, neoplasm /cancer genetics, oncogene Retroviridae, binding protein, chimeric protein, fusion gene, gene mutation, protein tyrosine kinase, stem cell, viral carcinogenesis laboratory mouse, transgenic animal
Project start date: 2001-04-01
Project end date: 2006-03-31
1R01CA090576-01 (2001): $353331
3R01CA090576-01S1 (2001): $18750
5R01CA090576-11 (2010): $286110
5R01CA090576-10 (2009): $286110
5R01CA090576-08 (2007): $286110
Sponsored Links Excellgen http://Excellgen.com
2R01CA090576-07 (2006): $295569
INHIBITOR OF THE C ABL TYROSINE KINASE
Richard A Van etten, Professor - Dept Of Medicine/hematology-
Immune Disease Institute, Inc.
800 Huntington Ave
boston, Ma 02115
Grant 5R01CA077691-04 from National Cancer Institute IRG: HEM
Abstract: c-Abl is a ubiquitously expressed non-receptor tyrosine kinase localized to the nucleus and actin cytoskeleton whose kinase activity is regulated in the stress response to genotoxic insults. The kinase activity of c-Abl appears to be negatively regulated in vivo by a cellular inhibitor binding via the Abl SH3 domain, and Abl is activated to become leukemogenic by several mechanisms, including SH3 deletion and chromosomal translocations in human leukemias. A yeast two-hybrid screen for Abl SH3-binding proteins identified a gene called PAG, whose protein product (Pag) has many properties expected of a physiological inhibitor of c-Abl. When co-expressed with c-Abl, Pag associates with Abl in vivo and potently suppresses the tyrosine phosphorylation induced by c-Abl, dependent on the Abl SH3 and kinase domains. Pag also inhibits the kinase activity of c-Abl in an immune complex kinase assay. However, purified Pag is a poor inhibitor of purified Abl, suggesting that additional proteins might be required. Interestingly, Pag has been previously identified as a member of a novel family of proteins induced by oxidative stress which have antioxidant and cell cycle regulatory properties. The long-term goals of this application are to further our understanding of the regulation and physiological function of c-Abl, and to better understand the mechanism of activation of leukemogenic forms of Abl. In the first aim, the biochemical mechanism of inhibition of c-Abl by Pag will be determined. The location of the Abl SH3 and kinase domain interaction sites in Pag will be mapped. The inhibition of Abl observed in vivo will be reconstituted in vitro using Pag as an affinity reagent to isolate c-Abl from cell extracts. The identity of an additional cellular protein required for in vitro inhibition will be pursued biochemically, and an additional two-hybrid screen against Pag will be carried out. It will be tested whether Pag has antioxidant activity and whether oxidative stress activates c-Abl. In the second aim, it will be determined whether Pag is a physiological inhibitor of c-Abl in vivo. The subcellular localization of Pag and whether endogenous Pag and Abl co-associate will be determined, and whether this association is regulated by DNA damage or oxidative stress. The function of endogenous Pag will be inhibited by antisense and dominant-negative strategies, and the consequences for Abl activation and phenotypic responses determined. It will be tested if overexpression of Pag can inhibit transformation by SH3-containing oncogenic Abl. These experiments will yield important new information about the function and regulation of c-Abl and the mode of activation of leukemogenic forms of Abl, and identify new avenues for anti-leukemic therapies
Keywords: enzyme activity, enzyme inhibitor, protein tyrosine kinase DNA damage, active site, antioxidant, binding protein, cell growth regulation, enzyme structure, epitope mapping, gene expression, ionizing radiation, oncogene, oxidative stress, phosphorylation 3T3 cell, SDS polyacrylamide gel electrophoresis, chimeric protein, flow cytometry, immunoprecipitation, mutant, protein purification, protein sequence, western blotting, yeast two hybrid system
Project start date: 1998-05-04
Project end date: 2003-02-28
5R01CA077691-04 (2001): $283674
5R01CA077691-03 (2000): $275408
3R01CA077691-02S1 (1999): $47300
5R01CA077691-02 (1999): $267386
1R01CA077691-01 (1998): $259598
REGULATION AND FUNCTION OF C AB1 NUCLEAR TYROSINE KINASE
Richard A Van etten, Professor - Dept Of Medicine/hematology-
Immune Disease Institute, Inc.
800 Huntington Ave
boston, Ma 02115
Grant 5R01CA072465-04 from National Cancer Institute IRG: CBY
Abstract: c-Abl is a nuclear tyrosine kinase whose function has remained elusive. It is reported to bind to DNA and bind to Rb and p53. The kinase activity is stimulated by DNA damage, leading to activation of the stress kinase pathway. The goal of this application is to understand the function of c-Abl as a growth inhibitor. By mutagenesis, specific functions of Abl require for the cytostatic response (DNA and p53 binding) will be tested. The effect o c-abl on inhibition of cyclins, cdk activity and CKIs will be examined and causality of the effects will be demonstrated by use of specific knock-out cells. Inducible expression will be used to define where in the abl effects maps within G1 phase. Finally, primary abl-/- fibroblasts (before and after rescue with selected abl constructs) will be used to assess the role of c-abl in growth arrest induced by DNA damage
Keywords: cell growth regulation, enzyme activity, growth inhibitor, protein structure /function, protein tyrosine kinase DNA binding protein, DNA damage, DNA directed RNA polymerase, cyclin, cyclin dependent kinase, phosphorylation, protein binding, regulatory gene, retinoblastoma protein, stress protein, tumor suppressor protein animal tissue, immunoprecipitation, ionizing radiation, polymerase chain reaction, site directed mutagenesis, western blotting
Project start date: 1998-05-04
Project end date: 2003-02-28
5R01CA072465-04 (2001): $231818
5R01CA072465-03 (2000): $223790
5R01CA072465-02 (1999): $217268
1R01CA072465-01A2 (1998): $210941
Sponsored Links Excellgen http://Excellgen.com
STUDIES OF BCR/ABL LEUKEMOGENESIS IN MICE
Richard A Van etten, Professor - Dept Of Medicine/hematology-
New England Medical Center Hospitals 750 Washington St Boston, Ma 021111533
Grant 5R01CA090576-06 from National Cancer Institute IRG: PTHB
Abstract: Adapted from the investigator s ) The human Philadelphia (Ph) chromosome-positive leukemias, including chronic myeloid leukemia (CML) and B-cell acute lymphoblastic leukemia, are among the most common hematological malignancies, and current therapy for these diseases is inadequate. Expression of the product of the t(9;22) Ph chromosome, the BCR!ABL fusion gene, in the hematopoietic system of mice by generation of transgenic mice or through retroviral transduction and transplantation of bone marrow has demonstrated that BCR/ABL is a leukemia specific oncogene and the direct cause of CML. The long term objective of this application is a more complete molecular and genetic understanding of the pathophysiology of human Ph-positive leukemias, particularly the myeloproliferative disease CML. These goals will be accomplished by the use of a retroviral bone marrow infection/transplantation mouse model system that accurately and quantitatively models both human CML and Ph-positive B-lymphoid leukemia, and will have two Specific Aims. In the first Aim, the signaling pathways important for leukemogenesis by BCR/ABL will be identified by testing BCR/ABL mutants, by further analysis of the requirement for direct binding of the Grb2 adapter protein by the Bcr/Abl fusion protein, and through the use of mice with germline mutations in signaling molecules. In the second Aim, the bone marrow target cells that initiate the CML-like disease and B-lymphoid leukemia induced by BCR/ABL will be characterized and isolated by physical and immunological methods. These investigations will add important new information to our understanding of these leukemias, that would be difficult if not impossible to obtain from studies in vitro, in cultured cells, or in primary human CML cells. This knowledge will be valuable for improving the diagnosis and treatment of the Ph-positive leukemias.
Keywords: acute lymphocytic leukemia, chromosome translocation, chronic myelogenous leukemia, neoplasm /cancer genetics, oncogene, Retroviridae, binding protein, chimeric protein, fusion gene, gene mutation, protein tyrosine kinase, stem cell, viral carcinogenesis, genetically modified animal, laboratory mouse
Project start date: 2001-04-01
Project end date: 2006-09-30
5R01CA090576-06 (2005): $338674
5R01CA090576-05 (2004): $296186
5R01CA090576-03 (2003): $393114
Pathogenesis & Therapy Of 8p11 Leukemia/Lymphoma
Richard A Van etten, Professor - Dept Of Medicine/hematology-
New England Medical Center Hospitals 750 Washington St Boston, Ma 021111533
Grant 5R01CA105043-04 from National Cancer Institute IRG: CAMP
Abstract: 8p11 stem cell leukemia/lymphoma syndrome, also known as 8p11 myeloproliferative syndrome (EMS), is a novel hematological malignancy characterized by chronic myeloproliferative disease, eosiniphilia, and non-Hodgkin s lymphoma. Current therapy for this disease is inadequate. The malignant cells from EMS patients have acquired chromosomal translocations involving the fibroblast growth factor receptor-1 (FGFR1) gene on 8p11 and express fusions of different N-terminal partner proteins with the tyrosine kinase domain of FGFR1. However, whether FGFR1 fusion proteins play a role in 8p11 syndrome and the molecular mechanisms underlying the pathogenesis of this disease are unknown. Recently, it has been demonstrated that different FGFR1 fusion proteins induce distinct leukemia/lymphoma syndromes in a mouse retroviral bone marrow transduction/transplantation model. These results implicate FGFR1 fusion tyrosine kinases as the direct cause of these malignancies and provide an accurate and quantitative animal model. In this application, this model system will be utilized to define the molecular pathogenesis of 8p11 leukemia/lymphoma syndrome and test targeted therapies for this disease. The first Aim will define the critical signaling pathways and the molecular mechanisms of induction of EMS-like disease in mice by the ZNF198-FGFR1 fusion tyrosine kinase, product of the (8;13) translocation in human 8pl 1 syndrome. This will be accomplished through biochemical analyses in Ba/F3 cells and primary leukemia cells from mice, studies with ZNF198-FGFR1 mutants, and use of mice with targeted mutations in signaling molecules. In the second Aim, the hematologic malignancies induced in mice by other FGFR1 fusions found in 8p11 syndrome, including FOP-FGFR1 and CEP110-FGFR1, will be characterized. The goal of the third Aim is preclinical testing of molecularly targeted therapeutic agents for 8p11 syndrome, comparing the antileukemic activity of several different FGFR1 kinase inhibitors in the mouse model of ZNF198-FGFR1- induced leukemia/lymphoma, and testing combinations of kinase inhibitors with drugs targeting essential downstream pathways identified in Aim 1. Collectively, these studies will yield important new knowledge about the pathogenesis and treatment of 8p11 syndrome that will also be valuable in extending targeted therapies to other hematologic malignancies and to solid tumors.
Keywords: blood /lymphatic pharmacology, blood disorder chemotherapy, leukemia, lymphoma, pathologic process, biological signal transduction, bone marrow transplantation, chromosome translocation, eosinophilia, fibroblast growth factor, gene expression, growth factor receptor, kinase inhibitor, phosphatidylinositol 3 kinase, phosphorylation, protein tyrosine kinase, flow cytometry, immunoprecipitation, laboratory mouse, polymerase chain reaction, western blotting
Project start date: 2004-04-01
Project end date: 2009-02-28
5R01CA105043-04 (2007): $316836
5R01CA105043-03 (2006): $326298
5R01CA105043-02 (2005): $334150
1R01CA105043-01 (2004): $332100
LEUKEMOGENESIS BY BCR/ABL GENES
Richard A Van etten, Professor - Dept Of Medicine/hematology-
Immune Disease Institute, Inc. 800 Huntington Ave Boston, Ma 02115
Grant 5R01CA057593-02 from National Cancer Institute IRG: HEM
Project start date: 1995-07-01
Project end date: 1998-06-30
5R01CA057593-02 (1996): $285459