E-Thesis 305 views 82 downloads
The Role of Ferroptosis in Tuberous Sclerosis Complex Disease / DOUGLAS FURLONG
Swansea University Author: DOUGLAS FURLONG
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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Swansea, Wales, UK
2025
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| Institution: | Swansea University |
| Degree level: | Master of Research |
| Degree name: | MRes |
| Supervisor: | Cronin, James ; Hitchings, Matthew |
| URI: | https://cronfa.swan.ac.uk/Record/cronfa69765 |
| 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 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. |
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| Keywords: |
Ferroptosis, tuberous sclerosis complex, Glutamine, GPX8, GLS2, SCD, RNA sequencing, CRISPR-Cas9 |
| College: |
Faculty of Medicine, Health and Life Sciences |