Grant-Serroukh, Dania;
(2019)
Lipid nanoparticles for mRNA-mediated cancer therapy.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
The use of messenger ribonucleic acid (mRNA) to introduce protein sequences into cells, expressing cytokines or antigens, is a promising therapeutic strategy in the oncology field. However, advancement of this technology is stalled by the lack of an effective delivery system. Developing mRNA-specific vectors, along with improving understanding of vector structure and fundamental cellular uptake mechanisms, should allow further progress in moving mRNA therapy to the clinic. This work investigates the use of lipid-peptide based vectors for the delivery of mRNA both in vitro and in vivo. The components of the vector were optimised to produce high levels of transfection in B16 melanoma cells. The resultant formulation comprised of the cationic lipid C14, the phospholipid DOPE, cholesterol and peptide 35. This formulation transfected a number of cancer cells types, producing high levels of expression of luciferase and green fluorescent protein. Formulations were characterised using dynamic light scattering, small-angle X-ray scattering, electron and atomic force microscopy to elucidate the structure of vectors. The nanocomplex had a spherical morphology and appeared to comprise an external lipid membrane with mRNA and/or peptides present on the surface. Optimal particles showed resistance to degradation and aggregation for up to 28 days at 4 oC and showed no loss of activity in the presence of RNase. Finally, mRNA uptake kinetics and trafficking were explored using fluorescent microscopy where no differences between optimal and non-optimal formulations were found; suggesting the key factor affecting performance was endosomal escape. Overall, the peptide-lipid-mRNA system was shown to be a promising platform for further investigation of mRNA delivery in vivo.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Lipid nanoparticles for mRNA-mediated cancer therapy |
| Event: | UCL |
| 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 > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > UCL School of Pharmacy |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10070394 |
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