Inducible human myeloid leukemic cell lines provide experimental access to events in both leukemic cell differentiation and normal hematopoiesis. It is possible that cytodifferentiation strategies could ultimately provide alternatives to ones based upon cytodestruction for the clinical management of human hematopoietic malignancies. Anti-sense RNA transfection analysis offers a means for directly assessing the functional role of certain genes, e.g., protooncogenes and colony stimulating factor (CSF) genes, in leukemic differentiation programs. Recent studies in our laboratory have established the utility of high copy number episomal replicons for the effective anti-sense RNA- mediated inhibition of gene expression in human hematopoietic cells. The proposed experiments are directed at: 1) optimization of episomal replicon-based constitutive and inducible transgene expression in human hematopoietic cells, 2) investigation of protooncogene and CSF mRNA expression in a panel of inducible human myeloid leukemic cell lines (HL-60, ML-1, K562, KG-1, U937), with specific attention to the chronology and lineage specificity of changes in gene expression, 3) anti-sense RNA mutational analysis to determine which, if any, expressed protooncogenes and CSFs play an obligatory role in leukemic differentiation, and 4) derivation of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) induction-specific HL-60 cDNAs through differential screening and cloning methodologies and functional evaluation of these genes through anti-sense RNA transfection experiments. The proposed studies should contribute to an understanding of the molecular mechanisms underlying normal and leukemic hematopoiesis, and could provide general insights into the effective use of anti-sense RNA technology in studying the role of specific genes in human hematopoietic cell function and differentiation.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
1R29CA047566-01
Application #
3458969
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1988-05-01
Project End
1993-04-30
Budget Start
1988-05-01
Budget End
1989-04-30
Support Year
1
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Case Western Reserve University
Department
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106
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Groger, R K; Morrow, D M; Tykocinski, M L (1989) Directional antisense and sense cDNA cloning using Epstein-Barr virus episomal expression vectors. Gene 81:285-94
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Hauer, C A; Getty, R R; Tykocinski, M L (1989) Epstein-Barr virus episome-based promoter function in human myeloid cells. Nucleic Acids Res 17:1989-2003
Kaplan, D R; Hambor, J E; Tykocinski, M L (1989) An immunoregulatory function for the CD8 molecule. Proc Natl Acad Sci U S A 86:8512-5
Hambor, J E; Tykocinski, M L; Kaplan, D R (1988) Functional consequences of anti-sense RNA-mediated inhibition of CD8 surface expression in a human T cell clone. J Exp Med 168:1237-45