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Inhibiting IL-17 Production by Blocking Endogenous RORγt Agonists for the Treatment of Autoimmune Diseases / DAPHNE DAVIES

Swansea University Author: DAPHNE DAVIES

DOI (Published version): 10.23889/SUthesis.60183

Abstract

The expression of pro-inflammatory cytokine IL-17 has been implicated in the pathogenesis of various autoimmune diseases. This project investigates the role of the cholesterol metabolising enzyme, CYP7B1 in the generation of the endogenous ligand of the master transcription factor, RORγt, driving th...

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Published: Swansea 2022
Institution: Swansea University
Degree level: Doctoral
Degree name: Ph.D
Supervisor: Griffiths, William J. ; Thornton, Cathy ; Mullins, Jonathan
URI: https://cronfa.swan.ac.uk/Record/cronfa60183
first_indexed 2022-06-13T09:40:56Z
last_indexed 2022-06-14T03:18:49Z
id cronfa60183
recordtype RisThesis
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spelling 2022-06-13T10:47:27.1667298 v2 60183 2022-06-13 Inhibiting IL-17 Production by Blocking Endogenous RORγt Agonists for the Treatment of Autoimmune Diseases 2b80d2b5284c458dc7f8353f2b92bfc2 DAPHNE DAVIES DAPHNE DAVIES true false 2022-06-13 The expression of pro-inflammatory cytokine IL-17 has been implicated in the pathogenesis of various autoimmune diseases. This project investigates the role of the cholesterol metabolising enzyme, CYP7B1 in the generation of the endogenous ligand of the master transcription factor, RORγt, driving the production of pro-inflammatory cytokine, IL-17 in Th17 cells. Oxysterols are oxidised derivatives of cholesterol which are intermediates in bile acid synthesis and have more recently been shown to have biological activity including roles in lipid homeostasis and immunity. This study uses an enzyme assisted derivatistion method with HPLC-ESI-MSn for free and total sterol analysis of CD4+ T cells from cell cultures isolated from healthy human donors. Inhibition of CYP7B1 activity with azole compounds is confirmed in vitro with Th17 differentiated cells isolated from healthy human donors and LC-MS analysis of oxysterols 24(S),25-diHC and 7α,24,25-triHC, upstream and downstream of CYP7B1 activity respectively. The activity of potent LXR agonist 24(S),25-EC, which is produced abundantly in CD4+ T cells, and downstream metabolites towards RORγt is investigated in vitro with CD4+ T cells from healthy human donors. The pathway in which 24(S),25-EC is deactivated in CD4+ T cells through CYP7B1 and HSD3B7 is revealed and the mechanism which determines agonists or inverse agonists activity of oxysterols towards RORγt is demonstrated through in silico molecular modelling. This study provides the foundations to progress the understanding of the role of oxysterols as RORγt ligands in Th17 cells. Further investigation may reveal a potential new target for the treatment of autoimmune diseases in which elevated Th17 cells and IL-17 production are observed. E-Thesis Swansea 26 2 2022 2022-02-26 10.23889/SUthesis.60183 COLLEGE NANME COLLEGE CODE Swansea University Griffiths, William J. ; Thornton, Cathy ; Mullins, Jonathan Doctoral Ph.D 2022-06-13T10:47:27.1667298 2022-06-13T10:37:34.5454859 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine DAPHNE DAVIES 1 60183__24290__81058c084d22444ba5b4bdab77cad04d.pdf Davies_Daphne_L_PhD_Thesis_Final_Redacted_Signature.pdf 2022-06-13T10:45:49.9404529 Output 17523947 application/pdf E-Thesis – open access true Copyright: The author, Daphne L. Davies, 2022. true eng
title Inhibiting IL-17 Production by Blocking Endogenous RORγt Agonists for the Treatment of Autoimmune Diseases
spellingShingle Inhibiting IL-17 Production by Blocking Endogenous RORγt Agonists for the Treatment of Autoimmune Diseases
DAPHNE DAVIES
title_short Inhibiting IL-17 Production by Blocking Endogenous RORγt Agonists for the Treatment of Autoimmune Diseases
title_full Inhibiting IL-17 Production by Blocking Endogenous RORγt Agonists for the Treatment of Autoimmune Diseases
title_fullStr Inhibiting IL-17 Production by Blocking Endogenous RORγt Agonists for the Treatment of Autoimmune Diseases
title_full_unstemmed Inhibiting IL-17 Production by Blocking Endogenous RORγt Agonists for the Treatment of Autoimmune Diseases
title_sort Inhibiting IL-17 Production by Blocking Endogenous RORγt Agonists for the Treatment of Autoimmune Diseases
author_id_str_mv 2b80d2b5284c458dc7f8353f2b92bfc2
author_id_fullname_str_mv 2b80d2b5284c458dc7f8353f2b92bfc2_***_DAPHNE DAVIES
author DAPHNE DAVIES
author2 DAPHNE DAVIES
format E-Thesis
publishDate 2022
institution Swansea University
doi_str_mv 10.23889/SUthesis.60183
college_str Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
document_store_str 1
active_str 0
description The expression of pro-inflammatory cytokine IL-17 has been implicated in the pathogenesis of various autoimmune diseases. This project investigates the role of the cholesterol metabolising enzyme, CYP7B1 in the generation of the endogenous ligand of the master transcription factor, RORγt, driving the production of pro-inflammatory cytokine, IL-17 in Th17 cells. Oxysterols are oxidised derivatives of cholesterol which are intermediates in bile acid synthesis and have more recently been shown to have biological activity including roles in lipid homeostasis and immunity. This study uses an enzyme assisted derivatistion method with HPLC-ESI-MSn for free and total sterol analysis of CD4+ T cells from cell cultures isolated from healthy human donors. Inhibition of CYP7B1 activity with azole compounds is confirmed in vitro with Th17 differentiated cells isolated from healthy human donors and LC-MS analysis of oxysterols 24(S),25-diHC and 7α,24,25-triHC, upstream and downstream of CYP7B1 activity respectively. The activity of potent LXR agonist 24(S),25-EC, which is produced abundantly in CD4+ T cells, and downstream metabolites towards RORγt is investigated in vitro with CD4+ T cells from healthy human donors. The pathway in which 24(S),25-EC is deactivated in CD4+ T cells through CYP7B1 and HSD3B7 is revealed and the mechanism which determines agonists or inverse agonists activity of oxysterols towards RORγt is demonstrated through in silico molecular modelling. This study provides the foundations to progress the understanding of the role of oxysterols as RORγt ligands in Th17 cells. Further investigation may reveal a potential new target for the treatment of autoimmune diseases in which elevated Th17 cells and IL-17 production are observed.
published_date 2022-02-26T05:55:29Z
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score 11.108426

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