title: miR-193a-3p interaction with HMGB1 downregulates human endothelial cell proliferation and migration.
abstract: Circulating endothelial colony forming cells (ECFCs) contribute to vascular repair where they are a target for therapy. Since ECFC proliferative potential is increased in cord versus peripheral blood and to define regulatory factors controlling this proliferation, we compared the miRNA profiles of cord blood and peripheral blood ECFC-derived cells. Of the top 25 differentially regulated miRNAs selected, 22 were more highly expressed in peripheral blood ECFC-derived cells. After validating candidate miRNAs by q-RT-PCR, we selected miR-193a-3p for further investigation. The miR-193a-3p mimic reduced cord blood ECFC-derived cell proliferation, migration and vascular tubule formation, while the miR-193a-3p inhibitor significantly enhanced these parameters in peripheral blood ECFC-derived cells. Using in silico miRNA target database analyses combined with proteome arrays and luciferase reporter assays of miR-193a-3p mimic treated cord blood ECFC-derived cells, we identified 2 novel miR-193a-3p targets, the high mobility group box-1 (HMGB1) and the hypoxia upregulated-1 (HYOU1) gene products. HMGB1 silencing in cord blood ECFC-derived cells confirmed its role in regulating vascular function. Thus, we show, for the first time, that miR-193a-3p negatively regulates human ECFC vasculo/angiogenesis and propose that antagonising miR-193a-3p in less proliferative and less angiogenic ECFC-derived cells will enhance their vasculo/angiogenic function.
authors.affiliation: Stem Cell Research, Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, University of Oxford, Oxford, OX3 9BQ, UK.|Stem Cell Research, NHS Blood and Transplant, Oxford, OX3 9BQ, UK.
authors.forename: Cheen P
authors.initials: CP
authors.lastname: Khoo
authors.affiliation: Stem Cell Research, Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, University of Oxford, Oxford, OX3 9BQ, UK.|Stem Cell Research, NHS Blood and Transplant, Oxford, OX3 9BQ, UK.|Laboratory of Biology, National and Kapodistrian University of Athens Medical School, Athens 115 27, Greece.|Cell and Gene Therapy Laboratory, Biomedical Research Foundation of the Academy of Athens (BRFAA), Athens, 11527, Greece.
authors.forename: Maria G
authors.initials: MG
authors.lastname: Roubelakis
authors.affiliation: Stem Cell Research, Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, University of Oxford, Oxford, OX3 9BQ, UK.|Stem Cell Research, NHS Blood and Transplant, Oxford, OX3 9BQ, UK.|Department of Biology, University of York, York, YO10 5DD, UK.
authors.forename: Jack B
authors.initials: JB
authors.lastname: Schrader
authors.affiliation: Stem Cell Research, Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, University of Oxford, Oxford, OX3 9BQ, UK.|Stem Cell Research, NHS Blood and Transplant, Oxford, OX3 9BQ, UK.|Institute of Molecular Biology and Biotechnology, Foundation of Research &Technology, GR-70013 Heraklion, Crete.
authors.forename: Grigorios
authors.initials: G
authors.lastname: Tsaknakis
authors.affiliation: Target Discovery Institute, NDM Research Building, Nuffield Department of Medicine, University of Oxford, OX3 7FZ, UK.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Konietzny
authors.affiliation: Target Discovery Institute, NDM Research Building, Nuffield Department of Medicine, University of Oxford, OX3 7FZ, UK.
authors.forename: Benedikt
authors.initials: B
authors.lastname: Kessler
authors.affiliation: The Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK.
authors.forename: Adrian L
authors.initials: AL
authors.lastname: Harris
authors.affiliation: Stem Cell Research, Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, University of Oxford, Oxford, OX3 9BQ, UK.|Stem Cell Research, NHS Blood and Transplant, Oxford, OX3 9BQ, UK.
authors.forename: Suzanne M
authors.initials: SM
authors.lastname: Watt
mesh_terms: D002465:Cell Movement / Q000502:physiology*; D049109:Cell Proliferation / Q000502:physiology*; D015536:Down-Regulation / Q000502:physiology*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D066026:Endothelial Progenitor Cells / Q000166:cytology / Q000378:metabolism; D024243:HMGB1 Protein / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*
chemical_list: D024243:HMGB1 Protein; C585491:HMGB1 protein, human; C546818:MIRN193 microRNA, human; D035683:MicroRNAs