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Title: Lipid nanoparticles for mRNA-mediated cancer therapy
Author: Grant-Serroukh, Dania
ISNI:       0000 0004 7964 908X
Awarding Body: UCL (University College London)
Current Institution: University College London (University of London)
Date of Award: 2019
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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.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available