Zahed Mohajerani, Safoura (2022) Establishment of a 3D cell culture perfused system to investigate mechanisms of blood vessel lumen morphogenesis: the role of cilia and cytoskeletal regulators in flow sensing. PhD thesis, University of Leeds.
Abstract
Cardiovascular function relies on stable and continuous formation of the blood vessel network by endothelial cells (ECs) to sustain blood flow and tissue viability. Although much is known regarding the molecules that control the initial stages of blood vessel growth, current understanding of the mechanisms by which the blood vessel lumen develops and expands to allow and sustain blood flow remain poorly elucidated. Endothelial primary cilia, microtubule-based organelles that protrude from the plasma membrane into the vessel lumen, mediate mechanotransduction of blood flow. Therefore, this thesis assesses the effects of genetic ablation of cilia, and cytoskeletal regulators DOCK4 and ROCK involved in ciliogenesis, on flow sensing and lumen formation under conditions of fluid flow.
A promising approach for understanding the mechanism behind development of a functional circulatory system is the use of perfused endothelial tubes in three- dimensional (3D) organotypic co-cultures of endothelial cells and fibroblasts. The system which was established in this study allowed manipulation of molecules associated with flow sensing under conditions of fluid flow. Molecular mechanisms of vascular lumen formation and lumen expansion were investigated using this system.
This study concludes that primary cilia are necessary for vascular tube formation in the co-culture model as their ablation inhibited lumen formation and expansion, and ROCK inhibition leading to disruption of proper cilium assembly inhibits vascular lumen formation. Furthermore, knockdown of GEF DOCK4 under conditions of FGF stimulation, increases angiogenesis in co-culture through higher EC proliferation accompanied by decreased cilia incidence. Finally, during this study a serotonin receptor HTR6-CFP2 fusion protein was cloned into a lentiviral vector and the protein was successfully expressed in ECs for assessment of ciliary dynamics in live cells.
Metadata
Supervisors: | MAVRIA, GEORGIA and JOHNSON, COLIN |
---|---|
Awarding institution: | University of Leeds |
Academic Units: | The University of Leeds > Faculty of Medicine and Health (Leeds) > School of Medicine (Leeds) |
Academic unit: | Leeds Institute of Medical Research (LIMR) |
Depositing User: | MS SAFOURA ZAHED MOHAJERANI |
Date Deposited: | 30 Jan 2023 11:57 |
Last Modified: | 01 Feb 2024 01:08 |
Open Archives Initiative ID (OAI ID): | oai:etheses.whiterose.ac.uk:32116 |
Download
Final eThesis - complete (pdf)
Filename: Zahed Mohajerani_SZM_MedicalScience_PhD_2022.pdf
Licence:
This work is licensed under a Creative Commons Attribution NonCommercial ShareAlike 4.0 International License
Export
Statistics
You do not need to contact us to get a copy of this thesis. Please use the 'Download' link(s) above to get a copy.
You can contact us about this thesis. If you need to make a general enquiry, please see the Contact us page.