The mechanisms regulating hematopoietic stem cell (HSC) proliferation, self-renewal, and differentiation are fundamental to our understanding and treatment of a number of hematopoietic disorders, including bone marrow failure syndromes and hematopoietic malignancies. Accumulating evidences suggests that stromal cells in the bone marrow provide key signals regulating HSC, yet the nature of these signals and even the stromal cell types that comprise the stem cell niche are poorly understood. Recent studies from our laboratory show that treatment with granulocyte colony-stimulating factor (G-CSF) results in marked changes in the bone marrow microenvironment that culminate in the mobilization of HSC into the blood. In this proposal, we plan to continue this research to answer the two following questions: 1) what are the cellular components of the stem cell niche;2) what are the signals that regulate the number and function of stem cell niche cells. This research has direct clinical relevance for stem cell transplantation, since it may lead to novel strategies to increase HSC mobilization yields and enhance HSC homing and engraftment following transplantation. Preliminary data support the hypothesis that factor(s) produced by cells of the monocyte lineage in the bone marrow provide key signals that regulate osteoblast function/survival and that expression of these factor(s) is regulated by G-CSF.
In Aim 1, we will identify candidate factors produced by monocytic cells and characterize their contribution to G-CSF-induced HSC mobilization and osteoblast suppression. The identification of factors that regulate osteoblasts may lead to the development of small molecule mimetics (or antagonists) that are able to stimulate (or inhibit) osteoblast (and possibly HSC) function. The chemokine Cxcl12 provides a key signal regulating HSC quiescence, survival, and trafficking. Within the bone marrow, Cxcl12 is expressed in osteoblasts, osteoblast precursors, endothelial cells, and Cxcl12- abundant reticular (CAR) cells.
In Aim 2, we will selectively delete Cxcl12 in selected stromal cel types and characterize their effect on HSC function and trafficking. To this end, we have generated transgenic mice carrying a floxed null allele of Cxcl12. These studies should provide new insight into the cellular composition of the stem cell niche and may lead to better strategies to augment HSC function in bone marrow failure syndromes or after stem cell transplantation. The following specific aims are proposed.
Aim 1. We will define the contribution of bone marrow monocytes/macrophages to HSC mobilization by G- CSF.
Aim 2. We will define the contribution of Cxcl12 expression by osteoblasts and endothelial cells to HSC function and trafficking.

Public Health Relevance

In the bone marrow, blood cells receive signals from stromal cells that regulate their growth, maturation, and release into the circulation. The proposed research is designed to better understand these signals. We believe this research will lead to novel strategies to mobilized blood stem cells for transplantation, enhance engraftment of stem cells after transplantation, and potential treat leukemia.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Project (R01)
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Molecular and Cellular Hematology (MCH)
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Mitchell, Phyllis
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Washington University
Internal Medicine/Medicine
Schools of Medicine
Saint Louis
United States
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Zhang, Jingzhu; Link, Daniel C (2016) Targeting of Mesenchymal Stromal Cells by Cre-Recombinase Transgenes Commonly Used to Target Osteoblast Lineage Cells. J Bone Miner Res 31:2001-2007
Rao, Mahil; Supakorndej, Teerawit; Schmidt, Amy P et al. (2015) Osteoclasts are dispensable for hematopoietic progenitor mobilization by granulocyte colony-stimulating factor in mice. Exp Hematol 43:110-4.e1-2
Calvi, Laura M; Link, Daniel C (2015) The hematopoietic stem cell niche in homeostasis and disease. Blood 126:2443-51
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Calvi, Laura M; Link, Daniel C (2014) Cellular complexity of the bone marrow hematopoietic stem cell niche. Calcif Tissue Int 94:112-24
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Schuettpelz, Laura G; Link, Daniel C (2013) Regulation of hematopoietic stem cell activity by inflammation. Front Immunol 4:204
Greenbaum, Adam; Hsu, Yen-Michael S; Day, Ryan B et al. (2013) CXCL12 in early mesenchymal progenitors is required for haematopoietic stem-cell maintenance. Nature 495:227-30
Schuettpelz, Laura G; Gopalan, Priya K; Giuste, Felipe O et al. (2012) Kruppel-like factor 7 overexpression suppresses hematopoietic stem and progenitor cell function. Blood 120:2981-9

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