title: Potential role for hyaluronan and the hyaluronan receptor RHAMM in mobilization and trafficking of hematopoietic progenitor cells.
abstract: 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(+)45(lo/med)Scatterlo/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 beta1 integrin and CD44, but not to RHAMM, whereas HPC motility was inhibited by MoAb to RHAMM and beta1 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.
authors.affiliation: Departments of Oncology and Medicine, University of Alberta, Cross Cancer Institute, Edmonton, Alberta, Canada. lpilarsk@gpu.srv.ualberta.ca
authors.forename: L M
authors.initials: LM
authors.lastname: Pilarski
authors.forename: E
authors.initials: E
authors.lastname: Pruski
authors.forename: J
authors.initials: J
authors.lastname: Wizniak
authors.forename: D
authors.initials: D
authors.lastname: Paine
authors.forename: K
authors.initials: K
authors.lastname: Seeberger
authors.forename: M J
authors.initials: MJ
authors.lastname: Mant
authors.forename: C B
authors.initials: CB
authors.lastname: Brown
authors.forename: A R
authors.initials: AR
authors.lastname: Belch
mesh_terms: D001781:Blood Component Removal; D001854:Bone Marrow Cells / Q000166:cytology / Q000473:pathology; D001943:Breast Neoplasms / Q000097:blood / Q000473:pathology; D002455:Cell Division; D002462:Cell Membrane / Q000502:physiology; D002465:Cell Movement; D016326:Extracellular Matrix Proteins / Q000502:physiology*; D005260:Female; D005786:Gene Expression Regulation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000473:pathology / Q000502:physiology*; D006801:Humans; D018960:Hyaluronan Receptors / Q000502:physiology*; D006820:Hyaluronic Acid / Q000235:genetics / Q000502:physiology*; D007700:Kinetics; D008223:Lymphoma / Q000097:blood / Q000473:pathology; D009101:Multiple Myeloma / Q000097:blood / Q000473:pathology; D012044:Regression Analysis
chemical_list: D016326:Extracellular Matrix Proteins; D018960:Hyaluronan Receptors; C098946:hyaluronan-mediated motility receptor; D006820:Hyaluronic Acid