Investigating the role of CLEC14A and RhoJ in regulating endothelial shear responses

Gill, Jagminder Singh (2023). Investigating the role of CLEC14A and RhoJ in regulating endothelial shear responses. University of Birmingham. M.Sc.

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Abstract

Atherosclerosis is a disease associated with dysregulated hemodynamic shear stress, manifesting in regions of low shear stress and disturbed flow, and contributing to an atheroprone phenotype. CLEC14A, a type I transmembrane glycoprotein, and RhoJ, a small Rho GTPase protein, are both endothelially expressed, involved in angiogenesis and regulated by shear stress. Both may have a role to play in regulating the atheroprotective transcription factor KLF2, which is a master regulator of shear stress responses such as endothelial cell alignment. The aim of this study was to gain insight into the mechanism through which CLEC14A regulates KLF2 expression, focusing on critical regions and interacting partners of CLEC14A. A second aim was to understand the role of RhoJ in regulating shear induced KLF2 expression and shear mediated cell alignment. A third aim was to enhance our understanding of CLEC14A and RhoJ protein biology by analysing bioinformatic transcriptome databases relating to shear stress, atherosclerosis, and endothelial signalling pathway studies. Subjecting HUVECs which were overexpressing CLEC14A to laminar shear stress suppressed the normal upregulation of KLF2 and enhanced MMRN2 expression. These effects were most pronounced with intact CLEC14A. RhoJ expression was negatively regulated by laminar shear stress and siRNA-mediated knockdown of RhoJ revealed no change in KLF2 expression. Key bioinformatic findings included the observation that both proteins could be regulated by EGR1, a transcription factor known to regulate and promote atherosclerosis. CLEC14A was downregulated by the proinflammatory cytokine TNF-α and RhoJ upregulated in a model of ischaemia-reperfusion injury. These studies have given additional insight into the biology of RhoJ and CLEC14A and the bioinformatic database analyses has yielded new avenues for future research.

Type of Work:

Thesis (Masters by Research > M.Sc.)

Award Type:

Masters by Research > M.Sc.

Supervisor(s):