Verbatim): Hematopoietic stem cells undergo a development stage-specific translocation during ontogeny and ultimately reside in the adult bone marrow. Maintenance of this highly regenerative cell pool through adult life is dependent upon their relative quiescence. We generated a cDNA library from quiescent human hematopoietic stem-like cells derived from bone marrow and identified by subtractive cloning a seven transmembrane molecule with a signature motif of the chemokine receptor family. Antiserum raised against this gene product identified cells from human fetal bone marrow, but not other fetal hematopoietic organs and very rare cells from adult bone marrow. These cells were enriched for quiescent cells with the ability to sustain mature blood cell generation for prolonged periods on stromal feeder layers. Calcium flux was induced upon exposure of transfected cells to bone marrow stroma conditioned medium, but not medium from other hematopoietic tissue stromal sources or from a panel of recombinant chemokines. Transduction of the receptor into hematopoietic cell lines resulted in enhanced transmigration toward bone marrow stroma in vitro and to bone marrow in irradiated mice. Transduced primary CD34+ progenitors had reduced proliferative potential, but sustained LTC-IC capability. Stem cell-G protein-coupled receptor-i (SC-GPR1) is a chemokine receptor that identifies bone marrow-derived hematopoietic stem cells and mediates growth regulatory and cell localization signals. This proposal builds upon these observations to address the following specific aims: 1. Define the functional role of SC-GPR1 in development using a mouse engineered to be deficient in the gene and its role in adult hematopoiesis using transplantation of cells identified by a monoclonal antibody specific for SC-GPR1. 2. Purify and clone the ligand for SC-GPR1. 3. Determine the mechanism by which ectopic expression of SC-GPR1 induces or maintains a primitive phenotype.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
2R01DK050234-07
Application #
6331287
Study Section
Hematology Subcommittee 2 (HEM)
Program Officer
Bishop, Terry Rogers
Project Start
1995-08-01
Project End
2005-03-31
Budget Start
2001-07-01
Budget End
2002-03-31
Support Year
7
Fiscal Year
2001
Total Cost
$337,842
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02199
Goncalves, Kevin A; Silberstein, Lev; Li, Shuping et al. (2016) Angiogenin Promotes Hematopoietic Regeneration by Dichotomously Regulating Quiescence of Stem and Progenitor Cells. Cell 166:894-906
Silberstein, Lev; Goncalves, Kevin A; Kharchenko, Peter V et al. (2016) Proximity-Based Differential Single-Cell Analysis of the Niche to Identify Stem/Progenitor Cell Regulators. Cell Stem Cell 19:530-543
Chattopadhyay, Shrikanta; Stewart, Alison L; Mukherjee, Siddhartha et al. (2015) Niche-Based Screening in Multiple Myeloma Identifies a Kinesin-5 Inhibitor with Improved Selectivity over Hematopoietic Progenitors. Cell Rep :
Scadden, David T (2014) Nice neighborhood: emerging concepts of the stem cell niche. Cell 157:41-50
Wang, Ying-Hua; Israelsen, William J; Lee, Dongjun et al. (2014) Cell-state-specific metabolic dependency in hematopoiesis and leukemogenesis. Cell 158:1309-1323
Kharchenko, Peter V; Silberstein, Lev; Scadden, David T (2014) Bayesian approach to single-cell differential expression analysis. Nat Methods 11:740-2
Sykes, Stephen M; Scadden, David T (2013) Modeling human hematopoietic stem cell biology in the mouse. Semin Hematol 50:92-100
Catic, André; Suh, Carol Y; Hill, Cedric T et al. (2013) Genome-wide map of nuclear protein degradation shows NCoR1 turnover as a key to mitochondrial gene regulation. Cell 155:1380-95
Kalaitzidis, Demetrios; Sykes, Stephen M; Wang, Zhu et al. (2012) mTOR complex 1 plays critical roles in hematopoiesis and Pten-loss-evoked leukemogenesis. Cell Stem Cell 11:429-39
Ferraro, Francesca; Lymperi, Stefania; Méndez-Ferrer, Simón et al. (2011) Diabetes impairs hematopoietic stem cell mobilization by altering niche function. Sci Transl Med 3:104ra101

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