The long-term objective of this project is to understand the basic nature of the regulation of hematopoiesis which in turn should provide a base for specific approaches to marrow failure states, to ameliorating the toxicity of various cytotoxic anti-cancer therapy and to leukemias involving the bone marrow.
The specific aims of this project are to continue to define cytokine production by a specific stromal cell line termed TC-1 evaluating the relationship of a previously described hemolymphopoietic growth factor (HLGF-1) to c-kit ligand, the biology of the high molecular weight forms of CSF-1 and the nature of a possibly unique B cell cytokine. We plan to continue studies on the production of cytokines by murine Dexter stromal cells with an emphasis on the functional significance of low level or subliminal cytokine production and to define the geographic and cellular localization of marrow cytokine production in Dexter stroma and in vivo models. We plan to clone the c-kit ligand from the TC-1 murine marrow stromal line using polymerase chain reaction techniques and to continue biochemical characterization of stromal derived CSF-1 and another possibly unique B cell factor. We will assess production of cytokines by stroma evaluating mRNA by standard Northern blot or utilizing polymerase chain amplification of reverse transcribed DNA as a more sensitive approach to detecting low level mRNA. We will also assess the production of biologically active protein by stromal cells and their subpopulations. We will focus on the biologic importance of low level factor production assessing factor dependent cell lines and purified hematopoietic progenitor/stem cells for their response to stromal derived growth factors either in conditioned media or when grown directly on stroma. We will evaluate the effects of blocking low level factor production with antibodies or antisense constructs. In the latter case, we will add antisense directly to cultures, transvect cultures with retroviral constructs with antisense linked to the metalothionine promoter or create transgenic murine models with growth factor antisense linked to the metalothionine. We will first evaluate interleukin-3, GM-CSF and c-kit ligand in these various models. Lastly, we plan to assess the geographic location of both transplanted stem cells and of cytokine producing cells in in vivo models and in in vitro Dexter culture using in situ hybridization.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA027466-16
Application #
3167660
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1979-07-01
Project End
1997-07-31
Budget Start
1993-08-20
Budget End
1994-07-31
Support Year
16
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of Massachusetts Medical School Worcester
Department
Type
Schools of Medicine
DUNS #
660735098
City
Worcester
State
MA
Country
United States
Zip Code
01655
Blomberg, M; Rao, S; Reilly, J et al. (1998) Repetitive bone marrow transplantation in nonmyeloablated recipients. Exp Hematol 26:320-4
Cooper, C L; Brady, G; Bilia, F et al. (1997) Expression of the Id family helix-loop-helix regulators during growth and development in the hematopoietic system. Blood 89:3155-65
Rao, S S; Peters, S O; Crittenden, R B et al. (1997) Stem cell transplantation in the normal nonmyeloablated host: relationship between cell dose, schedule, and engraftment. Exp Hematol 25:114-21
Kittler, E L; Peters, S O; Crittenden, R B et al. (1997) Cytokine-facilitated transduction leads to low-level engraftment in nonablated hosts. Blood 90:865-72
Reddy, G P; Quesenberry, P J (1996) Stem cell factor enhances interleukin-3 dependent induction of 68-kD calmodulin-binding protein and thymidine kinase activity in NFS-60 cells. Blood 87:3195-202
Quesenberry, P J; Iscove, N N; Cooper, C et al. (1996) Expression of basic helix-loop-helix transcription factors in explant hematopoietic progenitors. J Cell Biochem 61:478-88
Cao, Q P; McGrath, C A; Baril, E F et al. (1995) The 68 kDa calmodulin-binding protein is tightly associated with the multiprotein DNA polymerase alpha-primase complex in HeLa cells. Biochemistry 34:3878-83
Ramshaw, H S; Rao, S S; Crittenden, R B et al. (1995) Engraftment of bone marrow cells into normal unprepared hosts: effects of 5-fluorouracil and cell cycle status. Blood 86:924-9
Ramshaw, H S; Crittenden, R B; Dooner, M et al. (1995) High levels of engraftment with a single infusion of bone marrow cells into normal unprepared mice. Biol Blood Marrow Transplant 1:74-80
Stewart, F M; Temeles, D; Lowry, P et al. (1993) Post-5-fluorouracil human marrow: stem cell characteristics and renewal properties after autologous marrow transplantation. Blood 81:2283-9

Showing the most recent 10 out of 36 publications