{"id":{"repo_id":"wlv","oai_identifier":"oai:wlv.openrepository.com:2436/625918"},"canonical_url":"https://search.dev.ndltd.org/etd/wlv/oai:wlv.openrepository.com:2436/625918","repository":{"repo_id":"wlv","name":"University of Wolverhampton","base_url":"https://wlv.openrepository.com/server/oai/request"},"display":{"title":"Aberrant expression of mir-133a alters the expression of key inflammatory genes in the endothelium","abstract":"Introduction: Atherosclerosis is a major cause of cardiovascular diseases, driven by vascular inflammation that begins with endothelial dysfunction. The endothelium, which lines blood vessels, plays a critical role in regulating vascular tone and immune cell interactions. In response to inflammatory stimuli, such as cytokines, endothelial cells become activated, promoting the recruitment and adhesion of leukocytes to the vessel wall. This process, involving a complex sequence of leukocyte capture, rolling, and transmigration, contributes to the development of atherosclerotic lesions. Recent research has identified microRNAs as key regulators of endothelialleukocyte interactions, with miR-133a emerging as a potential modulator of this inflammatory response. This thesis explores the role of miR-133a in endothelial activation and leukocyte adhesion, aiming to uncover new therapeutic strategies to target these pathways in atherosclerosis. Methodology: Human endothelial cells were cultured in Endothelial Cell Growth Medium under aseptic conditions. Cells were transfected with miR-133a-3p or miR-133a-5p mimics and stimulated with TNF-α to induce an inflammatory response. To assess changes in gene expression, total RNA was extracted, quantified, and reverse-transcribed into cDNA. Quantitative real-time PCR (qRT-PCR) was then performed to measure the expression of key adhesion molecules and extracellular matrix proteins implicated in endothelial-leukocyte interactions. Statistical analyses were conducted using GraphPad Prism, employing parametric and non-parametric tests depending on data distribution. Additionally, bioinformatics tools, including miRWalk, TargetScan, and miRDB, were used to predict potential miR-133a target genes and evaluate their binding sites within the 3'UTR regions. Results: Both miR-133a-3p and miR-133a-5p downregulated ITGβ3, impairing endothelial cell migration. miR-133a-5p significantly reduced MMP-2 expression by 50%, contributing to endothelial barrier stabilization, while miR-133a-3p upregulated TIMP-3, enhancing barrier integrity. miR-133a-5p suppressed COL3A1 and TSP-1 expression, limiting vascular permeability and leukocyte recruitment. Furthermore, miR-133a-3p promoted ADAMTS-1 expression under TNF-α stimulation, supporting ECM remodelling, while miR-133a-3p also induced ICAM-1 expression in response to TNF-α, enhancing leukocyte adhesion. Conclusion: miR-133a regulates endothelial responses during inflammation. The distinct effects of its 3p and 5p strands on adhesion, matrix stability, and inflammatory signalling underscore miR- 133a’s potential as a therapeutic target in vascular inflammation and related pathologies.","abstract_html":"Introduction: Atherosclerosis is a major cause of cardiovascular diseases, driven by vascular inflammation that begins with endothelial dysfunction. The endothelium, which lines blood vessels, plays a critical role in regulating vascular tone and immune cell interactions. In response to inflammatory stimuli, such as cytokines, endothelial cells become activated, promoting the recruitment and adhesion of leukocytes to the vessel wall. This process, involving a complex sequence of leukocyte capture, rolling, and transmigration, contributes to the development of atherosclerotic lesions. Recent research has identified microRNAs as key regulators of endothelialleukocyte interactions, with miR-133a emerging as a potential modulator of this inflammatory response. This thesis explores the role of miR-133a in endothelial activation and leukocyte adhesion, aiming to uncover new therapeutic strategies to target these pathways in atherosclerosis. Methodology: Human endothelial cells were cultured in Endothelial Cell Growth Medium under aseptic conditions. Cells were transfected with miR-133a-3p or miR-133a-5p mimics and stimulated with TNF-α to induce an inflammatory response. To assess changes in gene expression, total RNA was extracted, quantified, and reverse-transcribed into cDNA. Quantitative real-time PCR (qRT-PCR) was then performed to measure the expression of key adhesion molecules and extracellular matrix proteins implicated in endothelial-leukocyte interactions. Statistical analyses were conducted using GraphPad Prism, employing parametric and non-parametric tests depending on data distribution. Additionally, bioinformatics tools, including miRWalk, TargetScan, and miRDB, were used to predict potential miR-133a target genes and evaluate their binding sites within the 3'UTR regions. Results: Both miR-133a-3p and miR-133a-5p downregulated ITGβ3, impairing endothelial cell migration. miR-133a-5p significantly reduced MMP-2 expression by 50%, contributing to endothelial barrier stabilization, while miR-133a-3p upregulated TIMP-3, enhancing barrier integrity. miR-133a-5p suppressed COL3A1 and TSP-1 expression, limiting vascular permeability and leukocyte recruitment. Furthermore, miR-133a-3p promoted ADAMTS-1 expression under TNF-α stimulation, supporting ECM remodelling, while miR-133a-3p also induced ICAM-1 expression in response to TNF-α, enhancing leukocyte adhesion. Conclusion: miR-133a regulates endothelial responses during inflammation. The distinct effects of its 3p and 5p strands on adhesion, matrix stability, and inflammatory signalling underscore miR- 133a’s potential as a therapeutic target in vascular inflammation and related pathologies.","abstract_has_math":false,"creators":["Kola, Teja Manidhar"],"institution":"University of Wolverhampton","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":"Research Institute in Healthcare Science, Faculty of Science and Engineering","school":null,"contributors":[],"advisors":["Aikaterini, Karakoula"],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025","date_published":"2025","updated_at":"2026-08-21T16:50:57Z","subjects":["miR-133a","gene expression","endothelial cells","inflammatory genes","MicroRNA","Atherosclerosis","vascular inflammation","endothelial dysfunction","regulatory mechanisms","cardiovascular disease"],"languages":["en"],"rights":[],"rights_urls":["https://wlv.openrepository.com/bitstreams/51fdc9aa-0cbe-4a89-a932-95dfe5dc5d9f/download"],"identifier_entries":[]},"links":{"outbound_url":"https://wlv.openrepository.com/handle/2436/625918","outbound_label":"Repository record","outbound_source":"dc:identifier.uri"},"source_record":{"url":"https://wlv.openrepository.com/server/oai/request?verb=GetRecord&metadataPrefix=dim&identifier=oai%3Awlv.openrepository.com%3A2436%2F625918","prefix":"dim"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Aikaterini, Karakoula"]},{"key":"dc:contributor.department","label":"Department","values":["Research Institute in Healthcare Science, Faculty of Science and Engineering"]},{"key":"dc:creator","label":"Author","values":["Kola, Teja Manidhar"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2025-04-24T10:10:01Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2025-04-24T10:10:01Z"]},{"key":"dc:date.issued","label":"Date","values":["2025"]},{"key":"dc:publisher","label":"Institution","values":["University of Wolverhampton"]},{"key":"dc:publisher.institution","label":"Dc Publisher Institution","values":["University of Wolverhampton"]},{"key":"dc:relation.isreferencedby","label":"Dc Relation Isreferencedby","values":["https://wlv.openrepository.com/handle/2436/625918"]},{"key":"dc:type","label":"Dc Type","values":["Thesis or dissertation"]},{"key":"dc:type.qualificationlevel","label":"Dc Type Qualificationlevel","values":["Masters"]},{"key":"dc:type.qualificationname","label":"Dc Type Qualificationname","values":["MPhil"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["miR-133a","gene expression","endothelial cells","inflammatory genes","MicroRNA","Atherosclerosis","vascular inflammation","endothelial dysfunction","regulatory mechanisms","cardiovascular disease"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["https://wlv.openrepository.com/bitstreams/51fdc9aa-0cbe-4a89-a932-95dfe5dc5d9f/download"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://wlv.openrepository.com/bitstreams/3dabdb48-6e28-48fd-a398-4c84adf8f208/download","https://wlv.openrepository.com/handle/2436/625918"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["A thesis submitted in partial fulfilment of the requirement of the University of Wolverhampton for the degree in Master of Philosophy."]},{"key":"dc:description.abstract","label":"Abstract","values":["Introduction: Atherosclerosis is a major cause of cardiovascular diseases, driven by vascular inflammation that begins with endothelial dysfunction. The endothelium, which lines blood vessels, plays a critical role in regulating vascular tone and immune cell interactions. In response to inflammatory stimuli, such as cytokines, endothelial cells become activated, promoting the recruitment and adhesion of leukocytes to the vessel wall. This process, involving a complex sequence of leukocyte capture, rolling, and transmigration, contributes to the development of atherosclerotic lesions. Recent research has identified microRNAs as key regulators of endothelialleukocyte interactions, with miR-133a emerging as a potential modulator of this inflammatory response. This thesis explores the role of miR-133a in endothelial activation and leukocyte adhesion, aiming to uncover new therapeutic strategies to target these pathways in atherosclerosis. Methodology: Human endothelial cells were cultured in Endothelial Cell Growth Medium under aseptic conditions. Cells were transfected with miR-133a-3p or miR-133a-5p mimics and stimulated with TNF-α to induce an inflammatory response. To assess changes in gene expression, total RNA was extracted, quantified, and reverse-transcribed into cDNA. Quantitative real-time PCR (qRT-PCR) was then performed to measure the expression of key adhesion molecules and extracellular matrix proteins implicated in endothelial-leukocyte interactions. Statistical analyses were conducted using GraphPad Prism, employing parametric and non-parametric tests depending on data distribution. Additionally, bioinformatics tools, including miRWalk, TargetScan, and miRDB, were used to predict potential miR-133a target genes and evaluate their binding sites within the 3'UTR regions. Results: Both miR-133a-3p and miR-133a-5p downregulated ITGβ3, impairing endothelial cell migration. miR-133a-5p significantly reduced MMP-2 expression by 50%, contributing to endothelial barrier stabilization, while miR-133a-3p upregulated TIMP-3, enhancing barrier integrity. miR-133a-5p suppressed COL3A1 and TSP-1 expression, limiting vascular permeability and leukocyte recruitment. Furthermore, miR-133a-3p promoted ADAMTS-1 expression under TNF-α stimulation, supporting ECM remodelling, while miR-133a-3p also induced ICAM-1 expression in response to TNF-α, enhancing leukocyte adhesion. Conclusion: miR-133a regulates endothelial responses during inflammation. The distinct effects of its 3p and 5p strands on adhesion, matrix stability, and inflammatory signalling underscore miR- 133a’s potential as a therapeutic target in vascular inflammation and related pathologies."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:format.checksum.md5","label":"Dc Format Checksum Md5","values":["42303f5f5d191cb96b8edf13eec650ff","79cc9c558089c2829a28510d0296eb06","b9e780bdde5f1967df91619ff2af0ba5"]},{"key":"dc:title","label":"Title","values":["Aberrant expression of mir-133a alters the expression of key inflammatory genes in the endothelium"]}]}],"canonical_facts":{"dc:contributor.advisor":["Aikaterini, Karakoula"],"dc:contributor.department":["Research Institute in Healthcare Science, Faculty of Science and Engineering"],"dc:creator":["Kola, Teja Manidhar"],"dc:date.accessioned":["2025-04-24T10:10:01Z"],"dc:date.available":["2025-04-24T10:10:01Z"],"dc:date.issued":["2025"],"dc:description":["A thesis submitted in partial fulfilment of the requirement of the University of Wolverhampton for the degree in Master of Philosophy."],"dc:description.abstract":["Introduction: Atherosclerosis is a major cause of cardiovascular diseases, driven by vascular inflammation that begins with endothelial dysfunction. The endothelium, which lines blood vessels, plays a critical role in regulating vascular tone and immune cell interactions. In response to inflammatory stimuli, such as cytokines, endothelial cells become activated, promoting the recruitment and adhesion of leukocytes to the vessel wall. This process, involving a complex sequence of leukocyte capture, rolling, and transmigration, contributes to the development of atherosclerotic lesions. Recent research has identified microRNAs as key regulators of endothelialleukocyte interactions, with miR-133a emerging as a potential modulator of this inflammatory response. This thesis explores the role of miR-133a in endothelial activation and leukocyte adhesion, aiming to uncover new therapeutic strategies to target these pathways in atherosclerosis. Methodology: Human endothelial cells were cultured in Endothelial Cell Growth Medium under aseptic conditions. Cells were transfected with miR-133a-3p or miR-133a-5p mimics and stimulated with TNF-α to induce an inflammatory response. To assess changes in gene expression, total RNA was extracted, quantified, and reverse-transcribed into cDNA. Quantitative real-time PCR (qRT-PCR) was then performed to measure the expression of key adhesion molecules and extracellular matrix proteins implicated in endothelial-leukocyte interactions. Statistical analyses were conducted using GraphPad Prism, employing parametric and non-parametric tests depending on data distribution. Additionally, bioinformatics tools, including miRWalk, TargetScan, and miRDB, were used to predict potential miR-133a target genes and evaluate their binding sites within the 3'UTR regions. Results: Both miR-133a-3p and miR-133a-5p downregulated ITGβ3, impairing endothelial cell migration. miR-133a-5p significantly reduced MMP-2 expression by 50%, contributing to endothelial barrier stabilization, while miR-133a-3p upregulated TIMP-3, enhancing barrier integrity. miR-133a-5p suppressed COL3A1 and TSP-1 expression, limiting vascular permeability and leukocyte recruitment. Furthermore, miR-133a-3p promoted ADAMTS-1 expression under TNF-α stimulation, supporting ECM remodelling, while miR-133a-3p also induced ICAM-1 expression in response to TNF-α, enhancing leukocyte adhesion. Conclusion: miR-133a regulates endothelial responses during inflammation. The distinct effects of its 3p and 5p strands on adhesion, matrix stability, and inflammatory signalling underscore miR- 133a’s potential as a therapeutic target in vascular inflammation and related pathologies."],"dc:format":["application/pdf"],"dc:format.checksum.md5":["42303f5f5d191cb96b8edf13eec650ff","79cc9c558089c2829a28510d0296eb06","b9e780bdde5f1967df91619ff2af0ba5"],"dc:identifier.uri":["https://wlv.openrepository.com/bitstreams/3dabdb48-6e28-48fd-a398-4c84adf8f208/download","https://wlv.openrepository.com/handle/2436/625918"],"dc:language.iso":["en"],"dc:publisher":["University of Wolverhampton"],"dc:publisher.institution":["University of Wolverhampton"],"dc:relation.isreferencedby":["https://wlv.openrepository.com/handle/2436/625918"],"dc:rights":["https://wlv.openrepository.com/bitstreams/51fdc9aa-0cbe-4a89-a932-95dfe5dc5d9f/download"],"dc:subject":["miR-133a","gene expression","endothelial cells","inflammatory genes","MicroRNA","Atherosclerosis","vascular inflammation","endothelial dysfunction","regulatory mechanisms","cardiovascular disease"],"dc:title":["Aberrant expression of mir-133a alters the expression of key inflammatory genes in the endothelium"],"dc:type":["Thesis or dissertation"],"dc:type.qualificationlevel":["Masters"],"dc:type.qualificationname":["MPhil"]},"updated_at":"2026-08-21T16:50:57Z"}