Successful hematopoietic transplantation relies on the capacity of intravenously infused hematopoietic stem cells (HSC) to home to and engraft bone marrow. This ability to home and migrate is central not only to the effective functioning of hematopoietic cells, but also to the development of the hematopoietic system. During vertebrate development, the site of hematopoieis moves from the yolk sac of the embryo to the embryo dorsal aorta to the fetal liver, then to the fetal spleen and finally to the bone marrow. Thus the HSC at each of these developmental stages must be able to migrate and home to location(s) that support their ability to self-renew and differentiate. Under this grant, we have made several surprising findings. First, we identified the homing receptors and chemokine receptors used by HSC to home from blood to marrow hematopoiesis sites. Next, we showed that fully competent, long term self-renewing HSC constitutively migrate through the blood and are capable of homing to niches, even in a parabiotic partner, and there establish long term hematopoiesis. Although ~100 HSC are in the blood of a mouse at any given time, their residence time in the blood, <5', is consistent with a daily flux of >104 HSC/mouse/day from marrow to blood to marrow. We now confirm that at any moment, up to 0.5% of marrow engraftment sites (niches) are open at a time, perhaps those vacated by the emigrating cells;these can accept donor HSC without conditioning. In SCID mice (and perhaps humans), single or repetitive (timed to match the opening of another 0.5% of niches) HSC infusions into unconditioned hosts gives significant engraftment and cures the immunodeficiency. In this grant we plan to pursue the specificity and nature of these engraftment sites and to better understand barriers to engraftment. We have developed an assay for the in vivo ectopic development and maintenance of bone marrow niches that will allow purification of the cells responsible for both niche development and niche maintenance;and have shown that modification of expression of particular proteins in these precursors can modify niche development and function. We will address the following specific aims: 1) To characterize the kinetics, magnitude, and therapeutic potential of HSC homing to and from BM sites under steady state conditions;2) To establish whether blockade of macrophage phagocytosis by CD47 enables mouse and human HSC homing to hematopoietic microenvironments and 3) To isolate and characterize HSC-supportive niche generating cells.

Public Health Relevance

Hematopoietic or blood forming stem cells (HSCs) are the only cells within the bone marrow (BM) that possess the ability to both differentiate to all blood lineages and to self-renew for life. These two properties allow for the proper maintenance of hematopoietic homeostasis. We are investigating another remarkable property of HSCs: their intrinsic ability to home upon intravenous transplantation to the highly specialized microenvironments within the BM that are required for them to function properly. It is because of this property that bone marrow or hematopoietic cell transplants successfully rescue patients from otherwise lethal damage to their blood system such as that experienced by cancer patients undergoing chemotherapy and radiation.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL058770-13
Application #
8426191
Study Section
Hematopoiesis Study Section (HP)
Program Officer
Thomas, John
Project Start
1998-04-01
Project End
2015-02-28
Budget Start
2013-03-01
Budget End
2015-02-28
Support Year
13
Fiscal Year
2013
Total Cost
$391,237
Indirect Cost
$155,617
Name
Stanford University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
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