Autophagy and formation of tubulovesicular autophagosomes provide a barrier against nonviral gene delivery

Roberts, Rebecca, Al-Jamal, Wafa, Whelband, Matthew, Thomas, Paul, Jefferson, Matthew, van den Bossche, Jeroen, Powell, Penny P, Kostarelos, Kostas and Wileman, Tom (2013) Autophagy and formation of tubulovesicular autophagosomes provide a barrier against nonviral gene delivery. Autophagy, 9 (5). pp. 667-682. ISSN 1554-8635

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    Abstract

    Cationic liposome (lipoplex) and polymer (polyplex)-based vectors have been developed for nonviral gene delivery. These vectors bind DNA and enter cells via endosomes, but intracellular transfer of DNA to the nucleus is inefficient. Here we show that lipoplex and polyplex vectors enter cells in endosomes, activate autophagy and generate tubulovesicular autophagosomes. Activation of autophagy was dependent on ATG5, resulting in lipidation of LC3, but did not require the PtdIns 3-kinase activity of PIK3C3/VPS34. The autophagosomes generated by lipoplex fused with each other, and with endosomes, resulting in the delivery of vectors to large tubulovesicular autophagosomes, which accumulated next to the nucleus. The tubulovesicular autophagosomes contained autophagy receptor protein SQSTM1/p62 and ubiquitin, suggesting capture of autophagy cargoes, but fusion with lysosomes was slow. Gene delivery and expression from both lipoplex and polyplex increased 8-fold in atg5 (-/-) cells unable to generate tubulovesicular autophagosomes. Activation of autophagy and capture within tubulovesicular autophagosomes therefore provides a new cellular barrier against efficient gene transfer and should be considered when designing efficient nonviral gene delivery vectors.

    Item Type: Article
    Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
    ?? MED-01 ??
    Faculty of Science > School of Pharmacy
    Faculty of Science > School of Biological Sciences
    University of East Anglia > Faculty of Medicine and Health Sciences > Research Groups > Gastroenterology and Gut Biology
    University of East Anglia > Faculty of Science > Research Groups > Drug Delivery and Pharmaceutical Materials
    Depositing User: Pure Connector
    Date Deposited: 28 Sep 2013 01:49
    Last Modified: 25 Jul 2018 08:38
    URI: https://ueaeprints.uea.ac.uk/id/eprint/43515
    DOI: 10.4161/auto.23877

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