Synthetic AAV/CRISPR vectors for blocking HIV-1 expression in persistently infected astrocytes

• Verfasst von: Kunze, Christine [VerfasserIn]
• Verfasst von: Börner, Kathleen [VerfasserIn]
• Verfasst von: Kienle, Eike [VerfasserIn]
• Verfasst von: Grimm, Dirk [VerfasserIn]
• Titel: Synthetic AAV/CRISPR vectors for blocking HIV-1 expression in persistently infected astrocytes
• Verf.angabe: Christine Kunze, Kathleen Börner, Eike Kienle, Tanja Orschmann, Ejona Rusha, Martha Schneider, Milena Radivojkov‐Blagojevic, Micha Drukker, Sabrina Desbordes, Dirk Grimm, Ruth Brack‐Werner
• E-Jahr: 2018
• Jahr: February 2018
• Umfang: 15 S.
• Fussnoten: First published: 09 November 2017 ; Gesehen am 05.11.2018
• Titel Quelle: Enthalten in: Glia
• Ort Quelle: Bognor Regis [u.a.] : Wiley-Liss, 1988
• Jahr Quelle: 2018
• Band/Heft Quelle: 66(2018), 2, Seite 413-427
• ISSN Quelle: 1098-1136
• DOI: doi:10.1002/glia.23254
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: adeno-associated viral vector
• Sach-SW: astrocytes
• Sach-SW: CRISPR/Cas9
• Sach-SW: HIV
• K10plus-PPN: 158257149X

Abstract

Astrocytes, the most abundant cells in the mammalian brain, perform key functions and are involved in several neurodegenerative diseases. The human immunodeficiency virus (HIV) can persist in astrocytes, contributing to the HIV burden and neurological dysfunctions in infected individuals. While a comprehensive approach to HIV cure must include the targeting of HIV-1 in astrocytes, dedicated tools for this purpose are still lacking. Here we report a novel Adeno-associated virus-based vector (AAV9P1) with a synthetic surface peptide for transduction of astrocytes. Analysis of AAV9P1 transduction efficiencies with single brain cell populations, including primary human brain cells, as well as human brain organoids demonstrated that AAV9P1 targeted terminally differentiated human astrocytes much more efficiently than neurons. We then investigated whether AAV9P1 can be used to deliver HIV-inhibitory genes to astrocytes. To this end we generated AAV9P1 vectors containing genes for HIV-1 proviral editing by CRISPR/Cas9. Latently HIV-1 infected astrocytes transduced with these vectors showed significantly diminished reactivation of proviruses, compared with untransduced cultures. Sequence analysis identified mutations/deletions in key HIV-1 transcriptional control regions. We conclude that AAV9P1 is a promising tool for gene delivery to astrocytes and may facilitate inactivation/destruction of persisting HIV-1 proviruses in astrocyte reservoirs.