Investigating the role of syncytiotrophoblast-derived extracellular vesicles in the pathogenesis of preeclampsia
<p><strong>Introduction:</strong> Preeclampsia (PE), a multi-systemic disorder of pregnancy affecting 2-8% of women globally, is one of the leading causes of maternal and neonatal morbidity and mortality. PE is diagnosed by <em>de novo</em> hypertension and end-organ da...
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| Other Authors: | |
| Format: | Thesis |
| Language: | English |
| Published: |
2024
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| _version_ | 1811140453114839040 |
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| author | Logenthiran, PV |
| author2 | Vatish, M |
| author_facet | Vatish, M Logenthiran, PV |
| author_sort | Logenthiran, PV |
| collection | OXFORD |
| description | <p><strong>Introduction:</strong> Preeclampsia (PE), a multi-systemic disorder of pregnancy affecting 2-8% of women globally, is one of the leading causes of maternal and neonatal morbidity and mortality. PE is diagnosed by <em>de novo</em> hypertension and end-organ damage. Although precise mechanisms are not fully elucidated, placenta is involved in the pathogenesis. Syncytiotrophoblast-derived extracellular vesicles (STB-EVs), which are membrane-bound biomolecules ranging between 30 – 1000 nm in size are gaining attention due to their diagnostic potential and their role in pathogenesis of PE. This thesis explores the roles of STB-EVs in pregnancy, focusing on the effects of their cargo (proteins and miRNA) on altering maternal physiology in PE and their potential to serve as non-invasive biomarkers. This research aimed to establish reliable protocols for isolating STB-EVs and to investigate their protein and genetic cargo, particularly focusing on kynurenine metabolising enzymes (KYNMEs) and microRNAs (miRNAs) relevant to preeclampsia.</p>
<p><strong>Aims:</strong> The study's objectives included isolating and characterizing STB-EVs, analysing their protein (kynurenine metabolising enzymes, KYNME) and genetic cargo (<em>hsa-miR-9-5p</em>), and exploring their functional implications PE.</p>
<p><strong>Methods:</strong> STB-EVs were isolated and enriched using ex vivo dual lobe placental perfusion and differential ultracentrifugation. Characterisation involved nanoparticle tracking analysis, western blotting, transmission electron microscopy, and flow cytometry. The protein cargo, particularly KYNMEs and their functionality, were analysed using immunohistochemistry, capillary electrophoresis and western blotting, and gas chromatography-mass spectrometry. Differentially expressed microRNAs were validated using quantitative polymerase chain reaction and their functional impacts were assessed <em>in vitro</em> on cerebral microvascular endothelial cells via assays for cell proliferation, viability and migration.</p>
<p><strong>Results:</strong> The isolation protocol successfully enriched STB-EVs. STB-EVs from normal and PE pregnancies contained KYNMEs with no significant differences between the two groups. Serum kynurenine levels were significantly lower in preeclamptic pregnancies. KYNMEs on STB-EVs demonstrated a dose-dependent attenuation of kynurenine levels <em>in vitro</em>. Additionally, differentially expressed miRNAs were validated, with <em>hsa-miR-9-5p</em> identified as a key target over-expressed in medium/large STB-EVs and serum EVs in preeclampsia. miR-9-5p attenuated cell proliferation and migration and altered the expression of angiogenesis-related proteins in cerebral microvascular endothelial cells.</p>
<p><strong>Conclusion:</strong> This thesis for the first time highlights the potential role of STB-EVs in kynurenine pathway and describes the anti-angiogenic effect of <em>hsa-miR-9-5p</em> on cerebral microvascular endothelial cells. The findings suggest that STB-EVs could be critical in regulating vascular and angiogenic responses in preeclampsia, providing a foundation for further understanding of the pathogenesis and future diagnostic strategies.</p> |
| first_indexed | 2024-09-25T04:22:13Z |
| format | Thesis |
| id | oxford-uuid:7a9705d2-6b45-40bf-b34c-335cf9bad441 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-09-25T04:22:13Z |
| publishDate | 2024 |
| record_format | dspace |
| spelling | oxford-uuid:7a9705d2-6b45-40bf-b34c-335cf9bad4412024-08-19T11:14:09ZInvestigating the role of syncytiotrophoblast-derived extracellular vesicles in the pathogenesis of preeclampsia Thesishttp://purl.org/coar/resource_type/c_db06uuid:7a9705d2-6b45-40bf-b34c-335cf9bad441PreeclampsiaEnglishHyrax Deposit2024Logenthiran, PVVatish, MTeixeira De Cerdeira, AS<p><strong>Introduction:</strong> Preeclampsia (PE), a multi-systemic disorder of pregnancy affecting 2-8% of women globally, is one of the leading causes of maternal and neonatal morbidity and mortality. PE is diagnosed by <em>de novo</em> hypertension and end-organ damage. Although precise mechanisms are not fully elucidated, placenta is involved in the pathogenesis. Syncytiotrophoblast-derived extracellular vesicles (STB-EVs), which are membrane-bound biomolecules ranging between 30 – 1000 nm in size are gaining attention due to their diagnostic potential and their role in pathogenesis of PE. This thesis explores the roles of STB-EVs in pregnancy, focusing on the effects of their cargo (proteins and miRNA) on altering maternal physiology in PE and their potential to serve as non-invasive biomarkers. This research aimed to establish reliable protocols for isolating STB-EVs and to investigate their protein and genetic cargo, particularly focusing on kynurenine metabolising enzymes (KYNMEs) and microRNAs (miRNAs) relevant to preeclampsia.</p> <p><strong>Aims:</strong> The study's objectives included isolating and characterizing STB-EVs, analysing their protein (kynurenine metabolising enzymes, KYNME) and genetic cargo (<em>hsa-miR-9-5p</em>), and exploring their functional implications PE.</p> <p><strong>Methods:</strong> STB-EVs were isolated and enriched using ex vivo dual lobe placental perfusion and differential ultracentrifugation. Characterisation involved nanoparticle tracking analysis, western blotting, transmission electron microscopy, and flow cytometry. The protein cargo, particularly KYNMEs and their functionality, were analysed using immunohistochemistry, capillary electrophoresis and western blotting, and gas chromatography-mass spectrometry. Differentially expressed microRNAs were validated using quantitative polymerase chain reaction and their functional impacts were assessed <em>in vitro</em> on cerebral microvascular endothelial cells via assays for cell proliferation, viability and migration.</p> <p><strong>Results:</strong> The isolation protocol successfully enriched STB-EVs. STB-EVs from normal and PE pregnancies contained KYNMEs with no significant differences between the two groups. Serum kynurenine levels were significantly lower in preeclamptic pregnancies. KYNMEs on STB-EVs demonstrated a dose-dependent attenuation of kynurenine levels <em>in vitro</em>. Additionally, differentially expressed miRNAs were validated, with <em>hsa-miR-9-5p</em> identified as a key target over-expressed in medium/large STB-EVs and serum EVs in preeclampsia. miR-9-5p attenuated cell proliferation and migration and altered the expression of angiogenesis-related proteins in cerebral microvascular endothelial cells.</p> <p><strong>Conclusion:</strong> This thesis for the first time highlights the potential role of STB-EVs in kynurenine pathway and describes the anti-angiogenic effect of <em>hsa-miR-9-5p</em> on cerebral microvascular endothelial cells. The findings suggest that STB-EVs could be critical in regulating vascular and angiogenic responses in preeclampsia, providing a foundation for further understanding of the pathogenesis and future diagnostic strategies.</p> |
| spellingShingle | Preeclampsia Logenthiran, PV Investigating the role of syncytiotrophoblast-derived extracellular vesicles in the pathogenesis of preeclampsia |
| title | Investigating the role of syncytiotrophoblast-derived extracellular vesicles in the pathogenesis of preeclampsia
|
| title_full | Investigating the role of syncytiotrophoblast-derived extracellular vesicles in the pathogenesis of preeclampsia
|
| title_fullStr | Investigating the role of syncytiotrophoblast-derived extracellular vesicles in the pathogenesis of preeclampsia
|
| title_full_unstemmed | Investigating the role of syncytiotrophoblast-derived extracellular vesicles in the pathogenesis of preeclampsia
|
| title_short | Investigating the role of syncytiotrophoblast-derived extracellular vesicles in the pathogenesis of preeclampsia
|
| title_sort | investigating the role of syncytiotrophoblast derived extracellular vesicles in the pathogenesis of preeclampsia |
| topic | Preeclampsia |
| work_keys_str_mv | AT logenthiranpv investigatingtheroleofsyncytiotrophoblastderivedextracellularvesiclesinthepathogenesisofpreeclampsia |