Institute of Virology, Helmholtz Center Munich, Neuherberg, 85764, Germany.
Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, 69120, Germany.
Glia. 2018 Feb;66(2):413-427. doi: 10.1002/glia.23254. Epub 2017 Nov 9.
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.
星形胶质细胞是哺乳动物大脑中最丰富的细胞，它们发挥着关键的功能，并参与多种神经退行性疾病。人类免疫缺陷病毒 (HIV) 可以在星形胶质细胞中持续存在，这导致了感染个体中 HIV 的负担和神经功能障碍。虽然实现 HIV 治愈的综合方法必须包括针对星形胶质细胞中的 HIV-1，但目前仍缺乏专门用于此目的的工具。在这里，我们报告了一种新型基于腺相关病毒的载体 (AAV9P1)，它带有用于星形胶质细胞转导的合成表面肽。对包括原代人类脑细胞和人类脑类器官在内的单细胞群体中的 AAV9P1 转导效率进行分析表明，AAV9P1 比神经元更有效地靶向终末分化的人类星形胶质细胞。然后，我们研究了 AAV9P1 是否可用于将 HIV 抑制基因递送至星形胶质细胞。为此，我们生成了包含用于通过 CRISPR/Cas9 进行 HIV-1 前病毒编辑的基因的 AAV9P1 载体。与未转导的培养物相比，用这些载体转导的潜伏性 HIV-1 感染的星形胶质细胞显示出前病毒的明显减少再激活。序列分析鉴定了关键 HIV-1 转录控制区中的突变/缺失。我们得出结论，AAV9P1 是一种将基因递送至星形胶质细胞的有前途的工具，并且可能有助于在星形胶质细胞储库中灭活/破坏持续存在的 HIV-1 前病毒。
