Campos De Souza, Senio;
(2019)
On the cellular responses to synthetic viruses.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
Despite almost five decades of pioneering work, the use of genes for therapeutic purposes has not yet been fully translated into clinical practice. While there has been some success with viral-based gene therapies, their broader use in the clinical setting remains limited due to safety concerns. For this reason, synthetic alternatives have been proposed, with a specific focus over polymeric and lipidic delivery systems. There is however a discrepancy between the transfection efficacy among different delivery systems even though the delivery efficiency of nucleic acids is similar across systems. The aims of the thesis herein were to explore DNA integrity following delivery and the potential role played by the cellular innate antiviral responses in tuning and controlling transfection. Efficacy of delivery and transfection with Lipofectamine® 2000, jetPEI® and polymersomes were studied. Confocal microscopy was employed to investigate DNA integrity following delivery within HEK293T cells. Cellular responses to nucleic acids were explored using NF-κB activation experiments as well as gene expression studies in a range of cells (HEK293T, FaDu, HeLa, A549, THP1 and HDF). Lastly, antiviral response inhibitors were used to explore the potential to improve transfection in various cells. All delivery systems were able to deliver the DNA into HEK293T and provided protection from degradation to some extent. The results also suggested that antiviral responses to nucleic acids were cell dependent, and these correlated with transfection efficacy. Namely, THP1 and HDF, two cell types difficult to transfect showed greatest NF-κB activation and upregulation of antiviral genes. The inhibition of specific antiviral pathways was able to improve transfection efficiency in certain conditions. Nevertheless, further studies are required to fully establish the link between antiviral response and successful gene transfection. The development of more efficient delivery systems would aid the development of a broader range of non-viral based therapeutics for gene therapy.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | On the cellular responses to synthetic viruses |
| Event: | UCL (University College London) |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2019. Original content in this thesis is licensed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) Licence (https://creativecommons.org/licenses/by/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request. |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences > Dept of Chemistry |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10081243 |
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