We are investigating the molecular mechanisms which control the individual and total concentrations of hemoglobins in human erythrocytes. In addition, we are studying the effects of functional alpha globin gene number, fetal hemoglobin (HbetaF) levels and the extent of red cell heterogeneity on the various manifestations of sickle cell disease and its genetic variants. The levels of each of the normal hemoglobins (A, A2, F) are determined by controls at the level of transcription and/or translation of the globin genes, as well as by factors that regulate protein degradation. The study of the control of hemoglobin levels has direct relevance to various hemoglobinopathies, especially thalassemia and sickle cell disease. In addition, these studies are of potential relevance to the more general question of control of gene expression in eukaryotic cells. For our experimental system, we are using the K562 human leukemic cell lines, as well as peripheral blood from individuals with sickle cell disease. We are studying the effects of short-term and long-term exposure of these cells to 5-azacytidine on their phenotype and the factors that control globin gene transcription. The cis-acting sequences in the promoter regions of the beta-like globin genes which interact with putative trans-acting factors are being delineated by a variety of molecular approaches. Concurrently, we are also attempting to develop a sickle cell mouse model by the introduction of a cloned human sickle globin gene into the mouse germ line by the microinjection of DNA into the pronuclei of fertilized eggs. The establishment of such a model would allow for basic and fundamental questions to be asked about the molecular, cellular and physiologic aspects of the disease, as well as provide as in vivo system to monitor the effects of potential therapy.

Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
1987
Total Cost
Indirect Cost
Name
U.S. National Inst Diabetes/Digst/Kidney
Department
Type
DUNS #
City
State
Country
United States
Zip Code