title: Characterization of a distinct population of circulating human non-adherent endothelial forming cells and their recruitment via intercellular adhesion molecule-3.
abstract: Circulating vascular progenitor cells contribute to the pathological vasculogenesis of cancer whilst on the other hand offer much promise in therapeutic revascularization in post-occlusion intervention in cardiovascular disease. However, their characterization has been hampered by the many variables to produce them as well as their described phenotypic and functional heterogeneity. Herein we have isolated, enriched for and then characterized a human umbilical cord blood derived CD133(+) population of non-adherent endothelial forming cells (naEFCs) which expressed the hematopoietic progenitor cell markers (CD133, CD34, CD117, CD90 and CD38) together with mature endothelial cell markers (VEGFR2, CD144 and CD31). These cells also expressed low levels of CD45 but did not express the lymphoid markers (CD3, CD4, CD8) or myeloid markers (CD11b and CD14) which distinguishes them from 'early' endothelial progenitor cells (EPCs). Functional studies demonstrated that these naEFCs (i) bound Ulex europaeus lectin, (ii) demonstrated acetylated-low density lipoprotein uptake, (iii) increased vascular cell adhesion molecule (VCAM-1) surface expression in response to tumor necrosis factor and (iv) in co-culture with mature endothelial cells increased the number of tubes, tubule branching and loops in a 3-dimensional in vitro matrix. More importantly, naEFCs placed in vivo generated new lumen containing vasculature lined by CD144 expressing human endothelial cells (ECs). Extensive genomic and proteomic analyses of the naEFCs showed that intercellular adhesion molecule (ICAM)-3 is expressed on their cell surface but not on mature endothelial cells. Furthermore, functional analysis demonstrated that ICAM-3 mediated the rolling and adhesive events of the naEFCs under shear stress. We suggest that the distinct population of naEFCs identified and characterized here represents a new valuable therapeutic target to control aberrant vasculogenesis.
authors.affiliation: Centre for Cancer Biology, South Australian Pathology, Adelaide, South Australia, Australia.
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Appleby
authors.forename: Michaelia P
authors.initials: MP
authors.lastname: Cockshell
authors.forename: Jyotsna B
authors.initials: JB
authors.lastname: Pippal
authors.forename: Emma J
authors.initials: EJ
authors.lastname: Thompson
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Barrett
authors.forename: Katie
authors.initials: K
authors.lastname: Tooley
authors.forename: Shaundeep
authors.initials: S
authors.lastname: Sen
authors.forename: Wai Yan
authors.initials: WY
authors.lastname: Sun
authors.forename: Randall
authors.initials: R
authors.lastname: Grose
authors.forename: Ian
authors.initials: I
authors.lastname: Nicholson
authors.forename: Vitalina
authors.initials: V
authors.lastname: Levina
authors.forename: Ira
authors.initials: I
authors.lastname: Cooke
authors.forename: Gert
authors.initials: G
authors.lastname: Talbo
authors.forename: Angel F
authors.initials: AF
authors.lastname: Lopez
authors.forename: Claudine S
authors.initials: CS
authors.lastname: Bonder
mesh_terms: D000071916:AC133 Antigen; D015703:Antigens, CD / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D002469:Cell Separation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D006023:Glycoproteins / Q000032:analysis; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D010455:Peptides / Q000032:analysis; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D013314:Stress, Mechanical; D015854:Up-Regulation
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D015815:Cell Adhesion Molecules; D006023:Glycoproteins; C483616:ICAM3 protein, human; C000606023:PROM1 protein, human; D010455:Peptides; D012333:RNA, Messenger