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Non-Viral Gene Therapy
A major collaborative research initiative has been established to investigate the structural requirements for lipids in a highly efficient ternary synthetic gene transfection system comprised of a lipid (L comprised of a cytofectin such as DOTMA and helper lipid DOPE)), targeting peptide (I) and DNA (D). This work is being carried out in collaboration with Dr.A.B.Tabor (Chemistry), Dr S. L.Hart (Institute of Child Health, UCL) and Professor M.J.Lawrence (Department of Pharmacy, KCL). These ternary lipopolyplex LID systems display high transfection efficiency and low toxicity in vitro and in vivo, transfect non-dividing cells efficiently, and are well tolerated with low immunogenicity in vivo. In this project we are tackling the major issues relating to vector efficacy including targeting, and the influence of the structure of the lipid on particle stability. We have established that shorter unsaturated lipid chains lead to enhanced transfection in several cell-types, most likely due to better membrane disruption or enhanced targeting, and that in the LD system the optical purity of the cytofectin may be important. We have also designed and synthesised two series of novel lipids, which enhance transfection efficacy due to better particle stability. In addition, we have studied the structure and fate of the aggregate formed using confocal microscopy and FCS (collaboration with Dr D. Zicha, ICRF) via the synthesis of specifically labelled lipids. Particle size, zeta potential and particle stability studies have been carried out. For in vivo applications, as well as incorporating PEG linkers to aid particle stability we have also introduced cleavable moieties to enhance release from the endosome.In a recent work on a collaborative EPSRC Nanotechnology Grand Challenge grant we are investigating the use of nanoparticles for the targeted delivery of therapeutic agents to the brain for the treatment of dementias. The aim is to deliver DNA, siRNA and therapeutic drugs to the brain using our targeted ternary delivery vectors comprised of designer lipids and peptides. Also, as part of the multidisciplinary King's College London and UCL Cancer Imaging Centre with Prof. T. Ng and 27 other scientists, we are developing new strategies for in vivo cancer imaging. The main clinical objective of the EPSRC/CRUK funded CCIC is to develop and provide a comprehensive range of imaging technologies which, when combined with molecular and genetic information, will begin to accurately characterize individual patients for the ultimate goal of personalized therapy.
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