About the Project
This project will generate human retinal ganglion cells (RGCs) for therapeutic testing. We will replace mouse tissue with human RGCs differentiated from patient-derived induced pluripotent stem cells (hiPSCs) from dominant optic atrophy caused by mutation in the OPA1 gene and from normal controls.
Dominant optic atrophy (ADOA) is one of the commonest mitochondrial optic neuropathies (MONs) caused by mutation in a nuclear mitochondrial gene, OPA1. There are currently no effective treatments, and patients become blind due to the death of RGCs, which take information from the eye to the brain via the optic nerves. Mitochondria are structures within the cell that convert food into cellular energy (ATP), with vision being of particularly high demand. This renders the visual system susceptible to mitochondrial disorders, including those that are genetic/inherited.
Many model systems, such as cells in culture, cells grown in 3D to resemble organs and whole organisms have been used to study disease. However, animal models may not be ideal as they do not model human disease precisely and pose ethical problems. Large numbers of mice and lengthy breeding is needed to obtain the required RGCs for analysis. As an alternative, OPA1 ADOA patient-derived cells can be reprogrammed back into stem cells, termed human induced pluripotent stem cell (hiPSC), and then turned into retinal cells including RGC. These human cellular models, offer many advantages, including: 1), better disease modelling as candidate therapies can be trialled on human ocular tissue; 2), significant replacement/reduction of mice culled for the procurement of primary RGCs for culture, because vast numbers of human RGCs can be generated; and 3), reduction of animals used for in vivo testing as the human culture system may be sufficient.
We hypothesise that improving mitochondrial function in RGCs differentiated from patient-derived iPSCs will result in increased RGC survival with preservation of normal cellular structure and function. We have developed a number of novel small molecule ubiquinone compounds, which tested in cell-based and tissue assays, demonstrate improved ATP rescue and reduced cell death. We will test whether our novel compounds are able to prevent cell death in this human patient model of the disease and promote normal cell structure and function. If we achieve this then we will have proof of principle to go forward to design testing in human patients.
This project offers the student the opportunity to acquired skills in iPSC culture, differentiation to RGCs and studies of mitochondrial structure and function. They will have experience of using iPSCs as a testing platform for therapeutic intervention and of transcriptomic analysis. The project is multi-disciplinary, and offer collaboration with The Welsh School of Pharmacy and Pharmaceutical Sciences and the Karolinska Institutet.
Applicants should apply to the Doctor of Philosophy in Vision Sciences with a start date of 1st October 2021.
In the research proposal section of your application, please specify the project title and supervisors of this project and copy the project description in the text box provided. In the funding section, please select the ‘self-funding’ option and specify the title of the studentship you are applying for. Please also include:
• an up-to-date CV
• a personal statement
• two references
We reserve the right to close the opportunity early if a sufficient number of suitable applications are received.
The successful candidate will begin the PhD/MPhil in Cardiff at the start of: October 2021
• Sun S, Erchova I, Sengpiel F, Votruba M. Opa1 deficiency leads to diminished mitochondrial bioenergetics with compensatory increased mitochondrial motility. 2020. Invest Ophthalmol Vis Sci. 61:42. doi: 10.1167/iovs.61.6.42
• 5. Williams PA, Morgan JE, Votruba M. Opa1 deficiency in a mouse model of dominant optic atrophy leads to retinal ganglion cell dendropathy. 2010. Brain. 133:2942
• 6. Williams PA, Piechota M, von Ruhland C, Taylor E, Morgan JE, Votruba M. Opa1 is essential for retinal ganglion cell synaptic architecture and connectivity. 2012. Brain. 135:493
• 7. Williams PA, Howell GR, Barbay JM, Braine CE, Sousa GL, John SW, Morgan JE. Retinal ganglion cell dendritic atrophy in DBA/2J glaucoma. 2013. PLoS One. 8: e72282
• 8. Williams PA, Tribble JR, Pepper KW, Cross SD, Morgan BP, Morgan JE, John SW, Howell GR. Inhibition of the classical pathway of the complement cascade prevents early dendritic and synaptic degeneration in glaucoma. 2016. Mol Neurodegen. 11:26
• 9. Varricchio C, Beirne K, Heard C, Newland B, Rozanowska M, Brancale A, Votruba M. The yin and yang of idebenone: Not too little, not too much – cell death in NQO1 deficient cells and the mouse retina. 2019. Free Radic Biol Med. doi: 10.1016/j.freeradbiomed.2019.11.030. PMID: 31775023
• 10. Chen et al., Modelling autosomal dominant optic atrophy using induced pluripotent stem cells and identifying potential therapeutic targets. 2016. Stem Cell Research & Therapy. 7: 2
• 11. Mead B, Chamling X; Zack DJ, Ahmed Z, Tomarev S. TNFα-Mediated Priming of Mesenchymal Stem Cells Enhances Their Neuroprotective Effect on Retinal Ganglion Cells. 2020. Investigative Ophthalmology & Visual Science. 61:6.
• 12. Mead B, Amaral J, Tomarev S. Mesenchymal Stem Cell–Derived Small Extracellular Vesicles Promote Neuroprotection in Rodent Models of Glaucoma. 2018. Invest Ophthalmol Vis Sci. 59: 702–714
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