T cell targeted nanoparticles for pulmonary siRNA delivery as novel asthma therapy

Kandil, Rima (2019): T cell targeted nanoparticles for pulmonary siRNA delivery as novel asthma therapy. Dissertation, LMU München: Fakultät für Chemie und Pharmazie

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.

• Dokumententyp: Dissertationen (Dissertation, LMU München)
• Keywords: siRNA, Pulmonary Delivery, Asthma, Lung, GATA3, T cells, Gene Knockdown, Nanoparticles, Polyplexes
• Themengebiete: 500 Naturwissenschaften und Mathematik ; 500 Naturwissenschaften und Mathematik > 540 Chemie
• Fakultäten: Fakultät für Chemie und Pharmazie
• Sprache der Hochschulschrift: Englisch
• Datum der mündlichen Prüfung: 7. November 2019
• 1. Bericht­erstatter:in: Merkel, Olivia