title: Hyaluronic acid and thrombin upregulate MT1-MMP through PI3K and Rac-1 signaling and prime the homing-related responses of cord blood hematopoietic stem/progenitor cells.
abstract: One of the hurdles of cord blood (CB) transplantation is delayed hematopoietic engraftment. Previously, we demonstrated that supernatants isolated from leukapheresis products of granulocyte-colony stimulating factor (G-CSF)-mobilized patients primed the homing of hematopoietic stem/progenitor cells (HSPC) by enhancing their chemotactic responses to stromal cell-derived factor (SDF)-1 and stimulating matrix metalloproteinases (MMPs) MMP-2 and MMP-9. Since membrane type 1 (MT1)-MMP activates proMMP-2 and localizes proteolytic activity at the leading edge of migrating cells, in this study we investigated whether MT1-MMP contributes to the priming of the homing-related responses of CB HSPC. We found that components of supernatants of leukapheresis products such as hyaluronic acid and thrombin (i) increase the secretion of proMMP-9 and transcription and protein synthesis of MT1-MMP in CB CD34(+) cells; (ii) increase the levels of active MMP-2 in co-cultures of CD34(+) cells with endothelial cells; (iii) increase the chemoinvasion across reconstituted basement membrane Matrigel of CD34(+) cells toward a low SDF-1 gradient (20 ng/mL); and (iv) activate mitogen-activated protein kinase, phosphatidylinositol 3-kinase, and Rac-1 signaling pathways. Inhibition of phosphatidylinositol 3-kinase and Rac-1 by their respective inhibitors LY290042 and NSC23766 attenuated MT1-MMP expression in CB CD34(+) cells, leading to reduced proMMP-2 activation and HSPC trans-Matrigel chemoinvasion toward SDF-1. Thus, our data suggest that MT1-MMP plays an important role in the homing-related responses of HSPC, and we propose that pretreatment of CB HSPC with hyaluronic acid or thrombin before transplantation could improve their homing and engraftment.
authors.affiliation: Canadian Blood Services R&D, Edmonton, Canada.
authors.forename: Neeta
authors.initials: N
authors.lastname: Shirvaikar
authors.forename: Leah A
authors.initials: LA
authors.lastname: Marquez-Curtis
authors.forename: Mariusz Z
authors.initials: MZ
authors.lastname: Ratajczak
authors.forename: Anna
authors.initials: A
authors.lastname: Janowska-Wieczorek
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D054377:Chemokine CXCL12 / Q000494:pharmacology; D018920:Coculture Techniques; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects; D005312:Fetal Blood / Q000166:cytology / Q000201:enzymology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D006801:Humans; D006820:Hyaluronic Acid / Q000494:pharmacology*; D042541:Intracellular Space / Q000201:enzymology / Q000378:metabolism; D007797:Laminin / Q000494:pharmacology; D053511:Matrix Metalloproteinase 14 / Q000378:metabolism*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D011509:Proteoglycans / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D013917:Thrombin / Q000494:pharmacology*; D015854:Up-Regulation / Q000187:drug effects*; D020830:rac1 GTP-Binding Protein / Q000378:metabolism*
chemical_list: D018952:Antigens, CD34; D054377:Chemokine CXCL12; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D006820:Hyaluronic Acid; D003094:Collagen; D019869:Phosphatidylinositol 3-Kinases; D013917:Thrombin; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; D053511:Matrix Metalloproteinase 14; D020830:rac1 GTP-Binding Protein