Investigating mechanisms of TDP-43 toxicity in cellular models of amyotrophic lateral sclerosis
Amyotrophic lateral sclerosis (ALS) is a fatal, neurodegenerative disorder for which there is currently no cure. TDP-43 cytoplasmic mislocalisation and aggregation are the pathological hallmarks of the disease, and mutations in the gene encoding TDP-43, TARDBP, have been observed in a small number o...
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| Formatua: | Thesis |
| Hizkuntza: | English |
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2022
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| _version_ | 1826316427065294848 |
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| author | Carroll, E |
| author2 | Talbot, K |
| author_facet | Talbot, K Carroll, E |
| author_sort | Carroll, E |
| collection | OXFORD |
| description | Amyotrophic lateral sclerosis (ALS) is a fatal, neurodegenerative disorder for which there is currently no cure. TDP-43 cytoplasmic mislocalisation and aggregation are the pathological hallmarks of the disease, and mutations in the gene encoding TDP-43, TARDBP, have been observed in a small number of ALS cases. Given this, it is hoped that modelling TARDBP mutations may provide early mechanistic insight into the disease process. Here, I examine the effects of the pathogenic TDP-43 M337V mutation on cellular phenotypes in an mESC-MN model of ALS. In TDP-43 M337V mESC-MNs, I observed reduced viability, both in unstressed and oxidative stressed conditions, and impaired stress granule formation in response to oxidative stress compared to TDP-43 WT controls. Interestingly, these changes were observed without evidence of TDP-43 cytoplasmic mislocalisation in TDP-43 M337V mESC-MNs when compared to TDP-43 WT controls. Increasing culture duration did not significantly alter synaptic protein expression or exacerbate phenotypes observed at earlier time points. Analysis of cellular energy metabolism revealed significantly reduced glycolysis in TDP-43 M337V mESC-MNs compared to TDP-43 WT controls in response to oxidative stress. Following oxidative stress, treatment with terazosin, which has previously been shown to significantly improve viability in our TDP-43 M337V mESC-MNs, significantly increased glycolysis whilst reducing mitochondrial respiration in TDP-43 M337V mESC-MNs. Finally, analysis of axonal transport in unstressed conditions using live imaging microscopy, revealed significantly impaired bi-directional transport of mitochondria and signalling endosomes in TDP-43 M337V mESC-MNs compared to TDP-43 WT controls. Overall, this work identifies relevant, ALS-associated phenotypes in the TDP-43 M337V mESC-MN model, which provides a valuable high-throughput system for investigating disease mechanisms and screening potential therapeutic compounds. |
| first_indexed | 2024-03-07T08:12:24Z |
| format | Thesis |
| id | oxford-uuid:3fbc216c-2cd2-4ff6-879b-9f99429ce5e6 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-12-09T03:44:56Z |
| publishDate | 2022 |
| record_format | dspace |
| spelling | oxford-uuid:3fbc216c-2cd2-4ff6-879b-9f99429ce5e62024-12-07T16:34:55ZInvestigating mechanisms of TDP-43 toxicity in cellular models of amyotrophic lateral sclerosisThesishttp://purl.org/coar/resource_type/c_db06uuid:3fbc216c-2cd2-4ff6-879b-9f99429ce5e6NeurosciencesEnglishHyrax Deposit2022Carroll, ETalbot, KCandalija, AMGordon, DAmyotrophic lateral sclerosis (ALS) is a fatal, neurodegenerative disorder for which there is currently no cure. TDP-43 cytoplasmic mislocalisation and aggregation are the pathological hallmarks of the disease, and mutations in the gene encoding TDP-43, TARDBP, have been observed in a small number of ALS cases. Given this, it is hoped that modelling TARDBP mutations may provide early mechanistic insight into the disease process. Here, I examine the effects of the pathogenic TDP-43 M337V mutation on cellular phenotypes in an mESC-MN model of ALS. In TDP-43 M337V mESC-MNs, I observed reduced viability, both in unstressed and oxidative stressed conditions, and impaired stress granule formation in response to oxidative stress compared to TDP-43 WT controls. Interestingly, these changes were observed without evidence of TDP-43 cytoplasmic mislocalisation in TDP-43 M337V mESC-MNs when compared to TDP-43 WT controls. Increasing culture duration did not significantly alter synaptic protein expression or exacerbate phenotypes observed at earlier time points. Analysis of cellular energy metabolism revealed significantly reduced glycolysis in TDP-43 M337V mESC-MNs compared to TDP-43 WT controls in response to oxidative stress. Following oxidative stress, treatment with terazosin, which has previously been shown to significantly improve viability in our TDP-43 M337V mESC-MNs, significantly increased glycolysis whilst reducing mitochondrial respiration in TDP-43 M337V mESC-MNs. Finally, analysis of axonal transport in unstressed conditions using live imaging microscopy, revealed significantly impaired bi-directional transport of mitochondria and signalling endosomes in TDP-43 M337V mESC-MNs compared to TDP-43 WT controls. Overall, this work identifies relevant, ALS-associated phenotypes in the TDP-43 M337V mESC-MN model, which provides a valuable high-throughput system for investigating disease mechanisms and screening potential therapeutic compounds. |
| spellingShingle | Neurosciences Carroll, E Investigating mechanisms of TDP-43 toxicity in cellular models of amyotrophic lateral sclerosis |
| title | Investigating mechanisms of TDP-43 toxicity in cellular models of amyotrophic lateral sclerosis |
| title_full | Investigating mechanisms of TDP-43 toxicity in cellular models of amyotrophic lateral sclerosis |
| title_fullStr | Investigating mechanisms of TDP-43 toxicity in cellular models of amyotrophic lateral sclerosis |
| title_full_unstemmed | Investigating mechanisms of TDP-43 toxicity in cellular models of amyotrophic lateral sclerosis |
| title_short | Investigating mechanisms of TDP-43 toxicity in cellular models of amyotrophic lateral sclerosis |
| title_sort | investigating mechanisms of tdp 43 toxicity in cellular models of amyotrophic lateral sclerosis |
| topic | Neurosciences |
| work_keys_str_mv | AT carrolle investigatingmechanismsoftdp43toxicityincellularmodelsofamyotrophiclateralsclerosis |