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T cell targeted nanoparticles for pulmonary siRNA delivery as novel asthma therapy
T cell targeted nanoparticles for pulmonary siRNA delivery as novel asthma therapy
The aim of this work was to develop and optimize a T cell targeted delivery system for pulmonary delivery of siRNA directed against GATA3, the central transcription factor of Th2 cytokines, as a novel therapy for asthma. Therefore, an existing carrier system on the basis of polycationic polymer polyethylenimine (PEI) and targeting Ligand transferrin (Tf), resulting in the so-called Tf-PEI, was chosen and fully characterized concerning relevant siRNA polyplex characteristics such as size, zeta potential, siRNA encapsulation efficiency and gene silencing capability in vitro and in vivo. Subsequently, Tf-PEI was blended with a second conjugate, Tf-Mel, containing the lysosomal Peptide melittin, in order to increase endosomal escape of the polyplexes. Resulting Tf-Mel-PEI blends were characterized and optimized to achieve siRNA polyplexes combining specific targeting of activated T cells and efficient cytoplasmic siRNA release, resulting in successful gene knockdown. For GATA3 silencing, a suitable siRNA sequence combination was found and applied within the Tf-Mel-PEI blend polyplexes to investigate down-stream effects of the gene knockdown on cytokine levels. These were concludingly tested in an optimized model for activated T cells as a first step for evaluation of relevant therapeutic effects in an inflammatory environment.
siRNA, Pulmonary Delivery, Asthma, Lung, GATA3, T cells, Gene Knockdown, Nanoparticles, Polyplexes
Kandil, Rima
2019
English
Universitätsbibliothek der Ludwig-Maximilians-Universität München
Kandil, Rima (2019): T cell targeted nanoparticles for pulmonary siRNA delivery as novel asthma therapy. Dissertation, LMU München: Faculty of Chemistry and Pharmacy
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Abstract

The aim of this work was to develop and optimize a T cell targeted delivery system for pulmonary delivery of siRNA directed against GATA3, the central transcription factor of Th2 cytokines, as a novel therapy for asthma. Therefore, an existing carrier system on the basis of polycationic polymer polyethylenimine (PEI) and targeting Ligand transferrin (Tf), resulting in the so-called Tf-PEI, was chosen and fully characterized concerning relevant siRNA polyplex characteristics such as size, zeta potential, siRNA encapsulation efficiency and gene silencing capability in vitro and in vivo. Subsequently, Tf-PEI was blended with a second conjugate, Tf-Mel, containing the lysosomal Peptide melittin, in order to increase endosomal escape of the polyplexes. Resulting Tf-Mel-PEI blends were characterized and optimized to achieve siRNA polyplexes combining specific targeting of activated T cells and efficient cytoplasmic siRNA release, resulting in successful gene knockdown. For GATA3 silencing, a suitable siRNA sequence combination was found and applied within the Tf-Mel-PEI blend polyplexes to investigate down-stream effects of the gene knockdown on cytokine levels. These were concludingly tested in an optimized model for activated T cells as a first step for evaluation of relevant therapeutic effects in an inflammatory environment.