title: Disruption of Microtubules Post-Virus Entry Enhances Adeno-Associated Virus Vector Transduction.
abstract: Perinuclear retention of viral particles is a poorly understood phenomenon observed during many virus infections. In this study, we investigated whether perinuclear accumulation acts as a barrier to limit recombinant adeno-associated virus (rAAV) transduction. After nocodazole treatment to disrupt microtubules at microtubule-organization center (MT-MTOC) after virus entry, we observed higher rAAV transduction. To elucidate the role of MT-MTOC in rAAV infection and study its underlying mechanisms, we demonstrated that rAAV's perinuclear localization was retained by MT-MTOC with fluorescent analysis, and enhanced rAAV transduction from MT-MTOC disruption was dependent on the rAAV capsid's nuclear import signals. Interestingly, after knocking down RhoA or inhibiting its downstream effectors (ROCK and Actin), MT-MTOC disruption failed to increase rAAV transduction or nuclear entry. These data suggest that enhancement of rAAV transduction is the result of increased trafficking to the nucleus via the RhoA-ROCK-Actin pathway. Ten-fold higher rAAV transduction was also observed by disrupting MT-MTOC in brain, liver, and tumor in vivo. In summary, this study indicates that virus perinuclear accumulation at MT-MTOC is a barrier-limiting parameter for effective rAAV transduction and defines a novel defense mechanism by which host cells restrain viral invasion.
authors.affiliation: 1 Gene Therapy Center, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina.|2 Cell and Developmental Biology, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina.
authors.forename: Ping-Jie
authors.initials: PJ
authors.lastname: Xiao
authors.affiliation: 1 Gene Therapy Center, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina.|3 Department of Microbiology and Immunology, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina.
authors.forename: Angela M
authors.initials: AM
authors.lastname: Mitchell
authors.affiliation: 4 Department of Statistics, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina.
authors.forename: Lu
authors.initials: L
authors.lastname: Huang
authors.affiliation: 1 Gene Therapy Center, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina.
authors.forename: Chengwen
authors.initials: C
authors.lastname: Li
authors.affiliation: 1 Gene Therapy Center, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina.|5 Department of Pharmacology, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina.
authors.forename: R Jude
authors.initials: RJ
authors.lastname: Samulski
mesh_terms: D000818:Animals; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000821:virology; D000229:Dependovirus / Q000378:metabolism*; D005822:Genetic Vectors / Q000378:metabolism*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D022101:Microtubule-Organizing Center / Q000187:drug effects / Q000378:metabolism; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism*; D015739:Nocodazole / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects; D014161:Transduction, Genetic; D053586:Virus Internalization / Q000187:drug effects
chemical_list: D015739:Nocodazole