title: miR-363-5p regulates endothelial cell properties and their communication with hematopoietic precursor cells.
abstract: Recent findings have shown that the blood vessels of different organs exert an active role in regulating organ function. In detail, the endothelium that aligns the vasculature of most organs is fundamental in maintaining organ homeostasis and in promoting organ recovery following injury. Mechanistically, endothelial cells (EC) of tissues such as the liver, lungs or the bone marrow (BM) have been shown to produce "angiocrine" factors that promote organ recovery and restore normal organ function. Controlled production of angiocrine factors following organ injury is therefore essential to promote organ regeneration and to restore organ function. However, the molecular mechanisms underlying the coordinated production and function of such "angiocrine" factors are largely undisclosed and were the subject of the present study. In detail, we identified for the first time a microRNA (miRNA) expressed by BM EC that regulates the expression of angiocrine genes involved in BM recovery following irradiation. Using a microarray-based approach, we identified several miRNA expressed by irradiated BMEC. After validating the variations in miRNA expression by semi-quantitative PCR, we chose to study further the ones showing consistent variations between experiments, and those predicted to regulate (directly or indirectly) angiogenic and angiocrine factors. Of the mi-RNA that were chosen, miR-363-5p (previously termed miR-363*) was subsequently shown to modulate the expression of numerous EC-specific genes including some angiocrine factors. By luciferase reporter assays, miR-363-5p is shown to regulate the expression of angiocrine factors tissue inhibitor of metalloproteinases-1 (Timp-1) and thrombospondin 3 (THBS3) at post-transcriptional level. Moreover, miR-363-5p reduction using anti-miR is shown to affect EC angiogenic properties (such as the response to angiogenic factors stimulation) and the interaction between EC and hematopoietic precursors (particularly relevant in a BM setting). miR-363-5p reduction resulted in a significant decrease in EC tube formation on matrigel, but increased hematopoietic precursor cells adhesion onto EC, a mechanism that is shown to involve kit ligand-mediated cell adhesion. Taken together, we have identified a miRNA induced by irradiation that regulates angiocrine factors expression on EC and as such modulates EC properties. Further studies on the importance of miR-363-5p on normal BM function and in disease are warranted.
authors.forename: Ana
authors.initials: A
authors.lastname: Costa
authors.forename: Joana
authors.initials: J
authors.lastname: Afonso
authors.forename: Catarina
authors.initials: C
authors.lastname: Osório
authors.forename: Ana L
authors.initials: AL
authors.lastname: Gomes
authors.forename: Francisco
authors.initials: F
authors.lastname: Caiado
authors.forename: Joana
authors.initials: J
authors.lastname: Valente
authors.forename: Sandra I
authors.initials: SI
authors.lastname: Aguiar
authors.forename: Francisco
authors.initials: F
authors.lastname: Pinto
authors.forename: Mário
authors.initials: M
authors.lastname: Ramirez
authors.affiliation: Angiogenesis Laboratory, Centro de Investigação em Patobiologia Molecular (CIPM), Instituto Português de Oncologia Francisco Gentil de Lisboa, EPE, Rua Professor Lima Basto, Lisbon 1099-023, Portugal. sergiodias@fm.ul.pt.
authors.forename: Sérgio
authors.initials: S
authors.lastname: Dias
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects; D002450:Cell Communication / Q000235:genetics*; D048708:Cell Growth Processes / Q000235:genetics; D042783:Endothelial Cells / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*
chemical_list: C574861:MIRN363 microRNA, human; D035683:MicroRNAs