): Erythropoiesis is a highly regulated developmental process during which pluripotent hematopoietic progenitor cells become committed to differentiate into erythroid precursors in response to erythropoietin and other growth factors. The presence of erythropoietin and c-Kit receptors is critical for this process to occur normally. It is not known, however, whether these receptors specifically """"""""instruct"""""""" progenitor cells to differentiate along a particular lineage or if they merely allow the cells to survive long enough so they can then differentiate along a stochastic program. This application seeks to address this issue in the physiological setting of the whole mouse, utilizing embryonic stem cell technology as well as the use of retroviruses to infect primary hematopoietic progenitors. The role of apoptosis will be examined by looking at the effect of expression of various a n ti-apoptotic genes such as Bcl-2, Bcl-x, CrmA, and c-IAPs on the differentiation of BaF3 and HCD-57 cell lines in addition to progenitors derived from fetal livers. Tandem mass spectrometry based protein sequencing approaches (MALDI-MS/MS) will be utilized to identify signaling molecules that a s s ociate with the EpoR when it is activated by Epo. Since the m o lecular/developmental defect in several red cell disorders such as polycythemia vera and erythroleukemias is not yet known, it is likely that these studies will lead to identification of molecules that may be defective in these diseases or provide insights into the developmental abnormality that leads to bone marrow hyperplasia. Finally, the nature of the signaling defect in polycythemia vera will be studied. It will be determined if the hyperresponsiveness of hematopoietic cells to Epo results from altered expression patterns or mutations affecting the function of negative modulators of signaling such as SH-PTP1 and CIS (Cytokine Inducible SH2 domain).

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
5K01CA075447-05
Application #
6376518
Study Section
Subcommittee G - Education (NCI)
Program Officer
Eckstein, David J
Project Start
1997-08-19
Project End
2002-03-01
Budget Start
2001-08-01
Budget End
2002-03-01
Support Year
5
Fiscal Year
2001
Total Cost
$72,045
Indirect Cost
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
Gronborg, Mads; Kristiansen, Troels Zakarias; Stensballe, Allan et al. (2002) A mass spectrometry-based proteomic approach for identification of serine/threonine-phosphorylated proteins by enrichment with phospho-specific antibodies: identification of a novel protein, Frigg, as a protein kinase A substrate. Mol Cell Proteomics 1:517-27
Pandey, Akhilesh; Dan, Ippeita; Kristiansen, Troels Z et al. (2002) Cloning and characterization of PAK5, a novel member of mammalian p21-activated kinase-II subfamily that is predominantly expressed in brain. Oncogene 21:3939-48
Jebanathirajah, Judith A; Peri, Suraj; Pandey, Akhilesh (2002) Toll and interleukin-1 receptor (TIR) domain-containing proteins in plants: a genomic perspective. Trends Plant Sci 7:388-91
Mann, Matthias; Ong, Shao En; Gronborg, Mads et al. (2002) Analysis of protein phosphorylation using mass spectrometry: deciphering the phosphoproteome. Trends Biotechnol 20:261-8
Steen, Hanno; Kuster, Bernhard; Fernandez, Minerva et al. (2002) Tyrosine phosphorylation mapping of the epidermal growth factor receptor signaling pathway. J Biol Chem 277:1031-9
Kratchmarova, Irina; Kalume, Dario E; Blagoev, Blagoy et al. (2002) A proteomic approach for identification of secreted proteins during the differentiation of 3T3-L1 preadipocytes to adipocytes. Mol Cell Proteomics 1:213-22
Blagoev, Blagoy; Nielsen, Mogens M; Angrist, Misha et al. (2002) Cloning of rat thymic stromal lymphopoietin receptor (TSLPR) and characterization of genomic structure of murine Tslpr gene. Gene 284:161-8
Pandey, Akhilesh; Blagoev, Blagoy; Kratchmarova, Irina et al. (2002) Cloning of a novel phosphotyrosine binding domain containing molecule, Odin, involved in signaling by receptor tyrosine kinases. Oncogene 21:8029-36
Pandey, Akhilesh; Ibarrola, Nieves; Kratchmarova, Irina et al. (2002) A novel Src homology 2 domain-containing molecule, Src-like adapter protein-2 (SLAP-2), which negatively regulates T cell receptor signaling. J Biol Chem 277:19131-8
Blagoev, Blagoy; Kratchmarova, Irina; Nielsen, Mogens M et al. (2002) Inhibition of adipocyte differentiation by resistin-like molecule alpha. Biochemical characterization of its oligomeric nature. J Biol Chem 277:42011-6

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