We will evaluate factors involved in the humoral and cellular modulation of hemopoiesis in normal individuals and those with hemopoietic diseases, including acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS). Major emphasis will be placed on evaluating (1) the in vitro interactions between purified recombinant human polypeptide hemopoietic growth: factors (HGFs) and human hemopoietic cells, and (2) the roles of enriched populations of accessory marrow and blood cells in generating HGFs under the influence of interleukin 2 (IL2) and other immune modulators. We are using a library of recombinant HGFs and monoclonal antibodies and have developed a variety of cell separation techniques, ligand binding procedures and serum-free cell proliferation, differentiation and clonogenic assays to perform these studies. Specific objectives regarding (1) hormonal interactions, are to determine, at the level of specific cell surface receptors, the role of the mesodermal stimulatory hormones insulin and insulin-like growth factors (IGFI & II) for modulating hemopoietic cell proliferation and differentiation, either directly or via alteration of receptor binding action for colony stimulating factor (GM-CSF) or erythropoietin. We will evaluate the hypothesis that multiple growth factors, including insulin and IGFs, interact in situ to provide cellular competence and progression factors for hemopoiesis. As hemopoietic progenitor cell (HPC) survival is partly mediated through HGF effects on glucose transport and ATP generation, we will examine the interactions between the metabolically active mitogenic hormones insulin, IGFs and CSFs to probe mechanisms of hemopoietic regulation. Studies using supplemented serum-free medium will be performed with human myeloid and erythroid cell lines and enriched HPCs as target cells to more precisely determine the functional effects and relationships between HGFs. Further, we will assess production of IGFs by these cells to evaluate the possible autocrine or paracrine roles of these hormones for leukemic vs normal cell growth. (2) We will examine the role of IL2 in enhancing provision of HGFs by enriched T and non T- cell subsets, with particular emphasis on the effector function of natural killer (NK) cells. We will assess production of proliferative (GM-CSF, BPA) and differentiative factors (G-CSF, DF) by resting T and NK cells (which lack TAC+ IL2 receptors) vs such activated Tac+) cells to evaluate the role of IL2 receptors in generating HGFs. After standardizing techniques to define HGF production, binding and responsiveness by normal myeloid cells and leukemic cell lines, we will (3) evaluate these parameters for cells from our patients with hemopoietic diseases, particularly AML and MDS, and will classify these disorders according to such basic cellular defects in growth regulation.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA036915-05
Application #
3174560
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1984-03-01
Project End
1990-11-30
Budget Start
1988-12-01
Budget End
1989-11-30
Support Year
5
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Stanford University
Department
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
Neubauer, A; Greenberg, P; Negrin, R et al. (1994) Mutations in the ras proto-oncogenes in patients with myelodysplastic syndromes. Leukemia 8:638-41
Greenberg, P L (1992) In vitro marrow culture studies in the myelodysplastic syndromes. Semin Oncol 19:34-46
Greenberg, P L; Negrin, R; Nagler, A (1990) Effects of CSFs in preleukemia. Bone Marrow Transplant 6 Suppl 1:121-6
Nagler, A; Ginzton, N; Negrin, R et al. (1990) Effects of recombinant human granulocyte colony stimulating factor and granulocyte-monocyte colony stimulating factor on in vitro hemopoiesis in the myelodysplastic syndromes. Leukemia 4:193-202
Greenberg, P; Negrin, R; Nagler, A et al. (1990) Effects of prolonged treatment of myelodysplastic syndromes with recombinant human granulocyte colony-stimulating factor. Int J Cell Cloning 8 Suppl 1:293-300;discussion 300-2
Greenberg, P L; Negrin, R; Nagler, A (1990) The use of haemopoietic growth factors in the treatment of myelodysplastic syndromes. Cancer Surv 9:199-212
Nagler, A; Greenberg, P L (1990) Bone marrow cell modulation and inhibition of myelopoiesis by large granular lymphocytes and natural killer cells. Int J Cell Cloning 8:171-83
Nagler, A; Binet, C; Mackichan, M L et al. (1990) Impact of marrow cytogenetics and morphology on in vitro hematopoiesis in the myelodysplastic syndromes: comparison between recombinant human granulocyte colony-stimulating factor (CSF) and granulocyte-monocyte CSF. Blood 76:1299-307
Lewinsohn, D M; Nagler, A; Ginzton, N et al. (1990) Hematopoietic progenitor cell expression of the H-CAM (CD44) homing-associated adhesion molecule. Blood 75:589-95
Greenberg, P; Negrin, R; Nagler, A et al. (1990) Effects of treatment of myelodysplastic syndromes with recombinant human granulocyte colony stimulating factor. Prog Clin Biol Res 338:151-61

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