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The Role of Ferroptosis in Tuberous Sclerosis Complex Disease / DOUGLAS FURLONG

Swansea University Author: DOUGLAS FURLONG

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Abstract

Tuberous Sclerosis Complex disease is a rare genetic disorder that results in the uncontrolled growth of affected cells, leading to the development of benign tumours in different tissues of the body that cause neurological disorders, skin problems, and heart, kidney and lung dysfunction. The conditi...

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Published: Swansea, Wales, UK 2025
Institution: Swansea University
Degree level: Master of Research
Degree name: MRes
Supervisor: Cronin, James ; Hitchings, Matthew
URI: https://cronfa.swan.ac.uk/Record/cronfa69765
first_indexed 2025-06-18T14:19:49Z
last_indexed 2025-06-19T10:47:23Z
id cronfa69765
recordtype RisThesis
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spelling 2025-06-18T15:30:31.7773451 v2 69765 2025-06-18 The Role of Ferroptosis in Tuberous Sclerosis Complex Disease b0fef9b8a7bcc8e2f4c4531a5403bd6c DOUGLAS FURLONG DOUGLAS FURLONG true false 2025-06-18 Tuberous Sclerosis Complex disease is a rare genetic disorder that results in the uncontrolled growth of affected cells, leading to the development of benign tumours in different tissues of the body that cause neurological disorders, skin problems, and heart, kidney and lung dysfunction. The condition is characterised by mutations in the TSC subunit genes (TSC1 or TSC2) leading to continuous activation of the mammalian target of rapamycin (mTOR) pathway, a central driver of cell proliferation and metabolism. Rapidly dividing cells require increased iron metabolism to maintain a wide spectrum of cellular processes, such as DNA synthesis and cellular respiration. However, these processes must be tightly regulated as excess iron is toxic to cells, via reactive oxygen species (ROS) and Fenton reactions, triggering ferroptosis, an iron-dependent form of programmed cell death. To avoid ferroptosis, cells must upregulate pathways that detoxify ROS. Tied into this is the role glutathione peroxidase enzymes (GPXs) play in regulating ROS homeostasis. Preliminary data from collaborators shows increased expression of Gpx8 in TSC2-/- mouse embryonic fibroblasts (MEFs). Localised to the endoplasmic reticulum (ER), GPX8 is an enzyme which neutralises the build-up of hydrogen peroxide (H2O2), an important ROS involved in lipid peroxidation and the induction of ferroptosis. Furthermore, the overexpression of GPX8 has been shown to be associated with an aggressive phenotype in breast cancer due to its influence on the epithelial-mesenchymal transition required for metastasis (Khatib et al., 2020). We hypothesise that GPX8 plays a role in the evasion of ferroptosis in Tuberous Sclerosis Complex patient tumours. This project will use RNA-seq data from TSC-/- AML cells to profile genes involved in ferroptosis. We will use TSC2-/- and TSC2+/+ (wt) angiomyolipoma (AML) cells derived from Tuberous Sclerosis Complex patient kidneys as a disease model to explore the role GPX8 and ferroptosis play in Tuberous Sclerosis Complex disease. The proteins involved in the induction/evasion of ferroptosis will be explored by immunoblotting, flow cytometry and confocal microscopy. Small molecule inducers of ferroptosis or siRNA targeting of GPX8 expression will be used for viability assays and wound healing assays in TSC2-/- vs wt cells. E-Thesis Swansea, Wales, UK Ferroptosis, tuberous sclerosis complex, Glutamine, GPX8, GLS2, SCD, RNA sequencing, CRISPR-Cas9 10 6 2025 2025-06-10 COLLEGE NANME COLLEGE CODE Swansea University Cronin, James ; Hitchings, Matthew Master of Research MRes 2025-06-18T15:30:31.7773451 2025-06-18T15:02:49.1620455 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Biomedical Science DOUGLAS FURLONG 1 69765__34513__38ebe1afb468468f9a009105ba0f8dbb.pdf Furlong_Douglas_MRes_Thesis_Final_Cronfa.pdf 2025-06-18T15:26:46.6173738 Output 2598585 application/pdf E-Thesis – open access true Copyright: The Author, Douglas Furlong, 2025. true eng
title The Role of Ferroptosis in Tuberous Sclerosis Complex Disease
spellingShingle The Role of Ferroptosis in Tuberous Sclerosis Complex Disease
DOUGLAS FURLONG
title_short The Role of Ferroptosis in Tuberous Sclerosis Complex Disease
title_full The Role of Ferroptosis in Tuberous Sclerosis Complex Disease
title_fullStr The Role of Ferroptosis in Tuberous Sclerosis Complex Disease
title_full_unstemmed The Role of Ferroptosis in Tuberous Sclerosis Complex Disease
title_sort The Role of Ferroptosis in Tuberous Sclerosis Complex Disease
author_id_str_mv b0fef9b8a7bcc8e2f4c4531a5403bd6c
author_id_fullname_str_mv b0fef9b8a7bcc8e2f4c4531a5403bd6c_***_DOUGLAS FURLONG
author DOUGLAS FURLONG
author2 DOUGLAS FURLONG
format E-Thesis
publishDate 2025
institution Swansea University
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 - Biomedical Science{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Biomedical Science
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description Tuberous Sclerosis Complex disease is a rare genetic disorder that results in the uncontrolled growth of affected cells, leading to the development of benign tumours in different tissues of the body that cause neurological disorders, skin problems, and heart, kidney and lung dysfunction. The condition is characterised by mutations in the TSC subunit genes (TSC1 or TSC2) leading to continuous activation of the mammalian target of rapamycin (mTOR) pathway, a central driver of cell proliferation and metabolism. Rapidly dividing cells require increased iron metabolism to maintain a wide spectrum of cellular processes, such as DNA synthesis and cellular respiration. However, these processes must be tightly regulated as excess iron is toxic to cells, via reactive oxygen species (ROS) and Fenton reactions, triggering ferroptosis, an iron-dependent form of programmed cell death. To avoid ferroptosis, cells must upregulate pathways that detoxify ROS. Tied into this is the role glutathione peroxidase enzymes (GPXs) play in regulating ROS homeostasis. Preliminary data from collaborators shows increased expression of Gpx8 in TSC2-/- mouse embryonic fibroblasts (MEFs). Localised to the endoplasmic reticulum (ER), GPX8 is an enzyme which neutralises the build-up of hydrogen peroxide (H2O2), an important ROS involved in lipid peroxidation and the induction of ferroptosis. Furthermore, the overexpression of GPX8 has been shown to be associated with an aggressive phenotype in breast cancer due to its influence on the epithelial-mesenchymal transition required for metastasis (Khatib et al., 2020). We hypothesise that GPX8 plays a role in the evasion of ferroptosis in Tuberous Sclerosis Complex patient tumours. This project will use RNA-seq data from TSC-/- AML cells to profile genes involved in ferroptosis. We will use TSC2-/- and TSC2+/+ (wt) angiomyolipoma (AML) cells derived from Tuberous Sclerosis Complex patient kidneys as a disease model to explore the role GPX8 and ferroptosis play in Tuberous Sclerosis Complex disease. The proteins involved in the induction/evasion of ferroptosis will be explored by immunoblotting, flow cytometry and confocal microscopy. Small molecule inducers of ferroptosis or siRNA targeting of GPX8 expression will be used for viability assays and wound healing assays in TSC2-/- vs wt cells.
published_date 2025-06-10T05:29:02Z
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score 11.444299

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