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Title: Biomimetic vectors for breast cancer iNOS gene therapy
Author: Zholobenko, Aleksey
Awarding Body: Queen's University Belfast
Current Institution: Queen's University Belfast
Date of Award: 2012
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Abstract:
While the field of gene delivery has matured dramatically in the last decade, there is still no gene delivery agent that can combine safety, efficacy and specificity into a single vehicle. In this work two protein based vectors, the Designer Biomimetic Vector (DBV) and RALA were investigated for the delivery of the gene coding for iNOS to breast cancer cells. Protein based vectors have the advantage of biocompatibility, bioderadabiliy and the high degree of control over the composition of the vector. They are not, however, widely studied at the present day. The DBV is a multi-domain recombinant chimeric protein, while RALA is a synthetic peptide with a single bi-functional domain. The DBV was found to form nanoparticles in the range of around 100mn and achieve a transfection efficacy of up to 10% in ZR-75-1 cells when complexed with pEG FP-NI. When complexed with a plasmid which allowed expression of iNOS under a constitutive CMV promoter (pENi), high concentrations of NO were produced (10μM) and near total cell kill was observed. Consequently, when the DBV/pEGP-N1 was tested in vivo, transfection was found in all organs and tissues tested, including tumour. Treatment with DBV/pENi nanoparticles resulted in a tumour growth delay of 17 days for both single dose and bi-weekly dose groups. Bi-weekly dosing was found to be toxic in an iNOS dependant manner. RALA was tested as an alternative vector and found ton form nanoparticles between 30-1 OOmn in diameter and give a transfection efficacy of up to 35% in ZR-75-1 cells when complexed with pEGFP-Nl. In in vivo models, RALAlpEGFP-N1 was found to transfect all organs and tissues tested, including tumour. In addition, in immunocompetant C57B mouse models, RALA was found to be no more immunogenic than mock injections while reducing the immunogenicity of plasmid DNA. RALA/pEGFP-N1 nanoparticles were found to protect the cargo DNA from both serum and freeze-drying. The relative efficacy and tolerability of both the DBV and RALA set aside a role for the two vehicles as prototypes for future targeted gene delivery systems
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.580137  DOI: Not available
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