Zusammenfassung
Genkonstrukte, die mit Polylysin-konjugierten Liganden wie Transferrin, EGF, oder Anti-CD 3-Antikörpern komplexiert sind, werden über Rezeptor-vermittelte Endozytose in Säugerzellen eingeschleust. Für einen effizienten Gentransfer müssen die Genkonstrukte aus der vesikulären Struktur der Endosomen in das Zytoplasma und weiter in den Zellkern transportiert werden. Als Endosomen-destabilisierende Komponenten wurden synthetische, von Influenzavirus- oder Rhinovirussequenzen abgeleitete Peptide oder ganze, inaktivierte Adenoviren eingesetzt. Als mögliche Anwendung des Gentransfersystems wird die Herstellung einer autologen, genmodifizierten Tumorvakzine zur Immunotherapie vorgestellt.
Summary
Gene constructs have been complexed with polylysine-conjugated ligands (such as transferrin, EGF, or anti-CD3 antibodies) for receptor-mediated uptake into endosomes. Accumulation of these complexes in intracellular vesicles however strongly reduces the efficiency of gene transfer. Viruses have acquired mechanisms to release their genome from endosomes into the cytoplasm. The endosomal acidification process specifically activates viral surface proteins that trigger desta-bilization of the endosomal membrane. This has led to the development of virus-like gene transfer complexes consisting of DNA complexed with a cell-binding ligand and polylysine-conjugated, endosomedisruption agents (such as defective adenoviruses, or small synthetic peptides derived from sequences of the influenza virus hemagglutinin or the rhinovirus VP-1 protein) which allow cytoplasmic entry of the DNA. DNA complexes linked to psoralen/UV-inactivated adenoviruses have been applied for the expression of high levels of cytokine genes in primary human cancer cells. The utility of the gene transfer system for the generation of gene-modified cancer vaccines is discussed.
Ich danke meinen Mitarbeitern Dr. Michael Buschle, Ing. Karl Mechtler, Dr. Berndt Oberhauser, Mag. Christian Plank, und Dr. Wolfgang Zauner. Die Arbeiten basieren in weiten Bereichen auf einer Zusammenarbeit mit den Gruppen von Prof. Max L. Birnstiel, Dr. Matt Cotten, Prof. Georg Stingl und Doz. Kurt Zatloukal.
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Literatur
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© 1995 Springer-Verlag Berlin Heidelberg
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Wagner, E. (1995). Rezeptorvermittelter Gentransfer Anwendung in der Tumorimmunotherapie?. In: Beger, H.G., Manns, M.P., Greten, H. (eds) Molekularbiologische Grundlagen der Gastroenterologie. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-79782-8_37
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DOI: https://doi.org/10.1007/978-3-642-79782-8_37
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