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Blood, Vol. 93 No. 9 (May 1), 1999:
pp. 2918-2927
Potential Role for Hyaluronan and the Hyaluronan Receptor RHAMM in
Mobilization and Trafficking of Hematopoietic Progenitor Cells
Linda M. Pilarski,
Eva Pruski,
Juanita Wizniak,
Darlene Paine,
Karen Seeberger,
Michael J. Mant,
Christopher B. Brown, and
Andrew
R. Belch
From the Departments of Oncology and Medicine, University of Alberta,
Cross Cancer Institute, Edmonton, Alberta, Canada; and the Department
of Medicine, University of Calgary, Calgary, Alberta, Canada.
Although the mechanism(s) underlying mobilization of hematopoietic
progenitor cells (HPCs) is unknown, detachment from the bone marrow
(BM) microenvironment and motility are likely to play a role. This
work analyzes the motile behavior of HPCs and the receptors
involved.
CD34+45lo/medScatterlo/med HPCs
from granulocyte colony-stimulating factor (G-CSF)-mobilized blood and
mobilized BM were compared with steady-state BM for their ability to
bind hyaluronan (HA), their expression of the HA receptors RHAMM and
CD44, and their motogenic behavior. Although RHAMM and CD44 are
expressed by mobilized blood HPCs, function blocking monoclonal
antibodies (MoAbs) identified RHAMM as a major HA binding receptor,
with a less consistent participation by CD44. Permeabilization of
mobilized blood HPCs showed a pool of intracellular (ic) RHAMM and a
smaller pool of icCD44. In contrast, steady-state BM HPCs have
significantly larger pools of icRHAMM and icCD44. Also, in contrast to
mobilized blood HPCs, for steady-state BM HPCs, MoAbs to RHAMM and CD44
act as agonists to upregulate HA binding. The comparison between
mobilized and steady-state BM HPCs suggests that G-CSF mobilization is
associated with depletion of intracellular stores of HA receptors and
modulates HA receptor usage. To confirm that mobilization alters the HA
receptor distribution and usage by HPCs, samples of BM were collected
at the peak of G-CSF mobilization in parallel with mobilized blood
samples. HA receptor distribution of mobilized BM HPCs was closely
matched with mobilized blood HPCs and different from steady-state BM
HPCs. Mobilized BM HPCs had lower pools of icHA receptors, similar to those of mobilized blood HPCs. Treatment of mobilized BM HPCs with
anti-RHAMM MoAb decreased HA binding, in contrast to steady-state BM
HPCs. Thus, G-CSF mobilization may stimulate an autocrine stimulatory loop for HPCs in which HA interacts with basal levels of RHAMM and/or
CD44 to stimulate receptor recycling. Consistent with this, treatment
of HPCs with azide, nystatin, or cytochalasin B increased HA binding,
implicating an energy-dependent process involving lipid rafts and the
cytoskeleton. Of the sorted HPCs, 66% were adherent and 27% were
motile on fibronectin plus HA. HPC adherence was inhibited by MoAbs to
 1 integrin and CD44, but not to RHAMM, whereas HPC motility was
inhibited by MoAb to RHAMM and 1 integrin, but not to CD44. This
finding suggests that RHAMM and CD44 play reciprocal roles in adhesion
and motility by HPCs. The G-CSF-associated alterations in RHAMM
distribution and the RHAMM-dependent motility of HPCs suggest a
potential role for HA and RHAMM in trafficking of HPCs and the possible
use of HA as a mobilizing agent in vivo.
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