title: The hematopoietic chemokine CXCL12 promotes integration of human endothelial colony forming cell-derived cells into immature vessel networks.
abstract: Proangiogenic factors, vascular endothelial growth factor (VEGF), and fibroblast growth factor-2 (FGF-2) prime endothelial cells to respond to "hematopoietic" chemokines and cytokines by inducing/upregulating expression of the respective chemokine/cytokine receptors. Coculture of human endothelial colony forming cell (ECFC)-derived cells with human stromal cells in the presence of VEGF and FGF-2 for 14 days resulted in upregulation of the "hematopoietic" chemokine CXCL12 and its CXCR4 receptor by day 3 of coculture. Chronic exposure to the CXCR4 antagonist AMD3100 in this vasculo/angiogenesis assay significantly reduced vascular tubule formation, an observation recapitulated by delayed AMD3100 addition. While AMD3100 did not affect ECFC-derived cell proliferation, it did demonstrate a dual action. First, over the later stages of the 14-day cocultures, AMD3100 delayed tubule organization into maturing vessel networks, resulting in enhanced endothelial cell retraction and loss of complexity as defined by live cell imaging. Second, at earlier stages of cocultures, we observed that AMD3100 significantly inhibited the integration of exogenous ECFC-derived cells into established, but immature, vascular networks. Comparative proteome profiler array analyses of ECFC-derived cells treated with AMD3100 identified changes in expression of potential candidate molecules involved in adhesion and/or migration. Blocking antibodies to CD31, but not CD146 or CD166, reduced the ECFC-derived cell integration into these extant vascular networks. Thus, CXCL12 plays a key role not only in endothelial cell sensing and guidance, but also in promoting the integration of ECFC-derived cells into developing vascular networks.
authors.affiliation: 1 Stem Cell Research Laboratory , Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, and NHS Blood and Transplant, John Radcliffe Hospital, Oxford, United Kingdom .
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Newey
authors.forename: Grigorios
authors.initials: G
authors.lastname: Tsaknakis
authors.forename: Cheen P
authors.initials: CP
authors.lastname: Khoo
authors.forename: Thanassi
authors.initials: T
authors.lastname: Athanassopoulos
authors.forename: Rosalba
authors.initials: R
authors.lastname: Camicia
authors.forename: Youyi
authors.initials: Y
authors.lastname: Zhang
authors.forename: Rita
authors.initials: R
authors.lastname: Grabowska
authors.forename: Adrian L
authors.initials: AL
authors.lastname: Harris
authors.forename: Maria G
authors.initials: MG
authors.lastname: Roubelakis
authors.forename: Suzanne M
authors.initials: SM
authors.lastname: Watt
mesh_terms: D001596:Benzylamines; D001808:Blood Vessels / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000378:metabolism*; D018920:Coculture Techniques / Q000379:methods; D000080027:Cyclams; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology*; D006413:Hematopoietic System / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology*; D006571:Heterocyclic Compounds / Q000494:pharmacology; D006801:Humans; D019718:Receptors, CXCR4 / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D001596:Benzylamines; C515255:CXCL12 protein, human; D054377:Chemokine CXCL12; D000080027:Cyclams; D006571:Heterocyclic Compounds; D019718:Receptors, CXCR4; D042461:Vascular Endothelial Growth Factor A; C088327:plerixafor