Please use this identifier to cite or link to this item: http://theses.ncl.ac.uk/jspui/handle/10443/4615
Full metadata record
DC FieldValueLanguage
dc.contributor.authorJordan, Nina-
dc.date.accessioned2020-01-16T16:39:08Z-
dc.date.available2020-01-16T16:39:08Z-
dc.date.issued2019-
dc.identifier.urihttp://theses.ncl.ac.uk/jspui/handle/10443/4615-
dc.descriptionPhD Thesisen_US
dc.description.abstractStudies in fibrotic diseases demonstrate that myofibroblasts may be derived from cell types other than fibroblasts. Sources include endothelial-to-mesenchymal transition (EndMT), generating 27% to 35% of myofibroblasts in cardiac fibrosis and around 10% in kidney fibrosis. Over the last decade, miRNAs have increasingly been described as key regulators in biologic processes but their profile remains mainly undescribed in EndMT. Therefore, we have investigated the possible role of miRNA signatures in EndMT models relevant to cardiac and kidney fibrosis. EndMT was modelled in human umbilical vein endothelial cells (HUVEC) by treatment with TGFβ2 (10ng/mL) and IL1β (10ng/mL). Significantly decreased expression of endothelial markers such as vWF and increased levels of mesenchymal markers such as fibronectin were observed by qPCR in HUVEC after 48 hours of treatment (p<0.05). Similarly, immunofluorescence showed increased expression of fibronectin and decreased expression of VE-cadherin in HUVEC 6 days post-treatment. In parallel, miRNAs were profiled with an nCounter assay in HUVEC. Profiles in untreated cells were compared to cells treated with TGFβ2 and IL1β. In these profiles, miR-126-3p was found down-regulated 24 hours’ post-treatment. Over-expression of miR-126-3p in HUVEC by transfection restored the expression of CD31 and repressed the expression of fibronectin induced by TGFβ2 and IL1β treatment, protecting the cells from EndMT induction. EndMT in vivo was investigated using lineage tracing with transgenic Cdh5-Cre-ERT2; Rosa26R-stop-YFP mice. Mice expressing YFP specifically in endothelial cells underwent myocardial infarction or unilateral ureteral obstruction. After sacrifice, lineage tracing showed expression of mesenchymal markers in endothelial derived cells, indicating the presence of EndMT in cardiac and kidney fibrosis. In addition, insitu hybridisation revealed the presence of miR-126-3p mainly in endothelial cells in mouse heart and kidney. We conclude that miR-126-3p may have a role in EndMT and this may have therapeutic potential in cardiac and kidney fibrosis.en_US
dc.description.sponsorshipfunded by Marie Curie Grant from the European Commission FP7 in the framework of the POSAT ITN (Prolong Organ Survival After Transplantation, Initial Training Networks, 606979)en_US
dc.language.isoenen_US
dc.publisherNewcastle Universityen_US
dc.titleIdentification and role of microRNAs in endothelial-to-mesenchymal transitionen_US
dc.typeThesisen_US
Appears in Collections:Institute of Cellular Medicine

Files in This Item:
File Description SizeFormat 
Jordan N 2019.pdfThesis180.33 MBAdobe PDFView/Open
dspacelicence.pdfLicence43.82 kBAdobe PDFView/Open


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.