Abstract

The human CD34 antigen is expressed specifically on human hematopoietic stem cells and not on mature blood cells, and CD34+ cells can reconstitute normal hematopoiesis of all cell lineages. CD34 is still the best defined human stem cell marker and is therefore a model for stem cell gene expression. Previous studies have defined some of the promoter elements for human and murine CD34, but the regulation of this and other stem cell genes is still very poorly understood, The distal elements necessary for human CD34 expression in vivo were recently isolated. The goal of this project is to continue to define and analyze the control elements of CD34, and to characterize those elements of the CD34 gene which can direct stem ceil expression of heterologous genes. The characterization of these elements could lead to the development of new vectors for gene therapy, as well as new understanding of the factors regulating genes in stem cells. Therefore, the specific aims are to characterize and study the regulatory elements of CD34 in the following steps: 1. To further characterize the important control elements regulating human CD34 gene expression in stem cells. 2. To characterize DNA binding proteins which regulate these elements. 3. To use CD34 regulatory elements expressing Cre recombinase and the tetracycline transactivator to enable tissue specific, regulatable expression of heterologous genes in the stem cell compartment.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK048660-12
Application #
6985374
Study Section
Hematology Subcommittee 2 (HEM)
Program Officer
Bishop, Terry Rogers
Project Start
1994-09-30
Project End
2007-11-30
Budget Start
2005-12-01
Budget End
2006-11-30
Support Year
12
Fiscal Year
2006
Total Cost
$385,444
Indirect Cost

  Institution

Name
Beth Israel Deaconess Medical Center
Department
Type
DUNS #
071723621
City
Boston
State
MA
Country
United States
Zip Code
02215

  Publications

Koschmieder, Steffen; Gottgens, Berthold; Zhang, Pu et al. (2005) Inducible chronic phase of myeloid leukemia with expansion of hematopoietic stem cells in a transgenic model of BCR-ABL leukemogenesis. Blood 105:324-34
Huettner, Claudia S; Koschmieder, Steffen; Iwasaki, Hiromi et al. (2003) Inducible expression of BCR/ABL using human CD34 regulatory elements results in a megakaryocytic myeloproliferative syndrome. Blood 102:3363-70
Okuno, Yutaka; Huettner, Claudia S; Radomska, Hanna S et al. (2002) Distal elements are critical for human CD34 expression in vivo. Blood 100:4420-6
Radomska, Hanna S; Gonzalez, David A; Okuno, Yutaka et al. (2002) Transgenic targeting with regulatory elements of the human CD34 gene. Blood 100:4410-9
Radomska, H S; Satterthwaite, A B; Taranenko, N et al. (1999) A nuclear factor Y (NFY) site positively regulates the human CD34 stem cell gene. Blood 94:3772-80
Radomska, H S; Satterthwaite, A B; Burn, T C et al. (1998) Multiple control elements are required for expression of the human CD34 gene. Gene 222:305-18
Iwama, A; Zhang, P; Darlington, G J et al. (1998) Use of RDA analysis of knockout mice to identify myeloid genes regulated in vivo by PU.1 and C/EBPalpha. Nucleic Acids Res 26:3034-43
Yamanaka, R; Kim, G D; Radomska, H S et al. (1997) CCAAT/enhancer binding protein epsilon is preferentially up-regulated during granulocytic differentiation and its functional versatility is determined by alternative use of promoters and differential splicing. Proc Natl Acad Sci U S A 94:6462-7
Tenen, D G; Hromas, R; Licht, J D et al. (1997) Transcription factors, normal myeloid development, and leukemia. Blood 90:489-519
Hohaus, S; Petrovick, M S; Voso, M T et al. (1995) PU.1 (Spi-1) and C/EBP alpha regulate expression of the granulocyte-macrophage colony-stimulating factor receptor alpha gene. Mol Cell Biol 15:5830-45

Showing the most recent 10 out of 12 publications