CCR5 Gene Editing of Resting CD4+ T Cells by Transient ZFN Expression From HIV Envelope Pseudotyped Nonintegrating Lentivirus Confers HIV-1 Resistance in Humanized Mice

• Main Authors: Guohua Yi, Jang Gi Choi, Preeti Bharaj, Sojan Abraham, Ying Dang, Tal Kafri, Ogechika Alozie, Manjunath N Manjunath, Premlata Shankar
• Format: Article
• Language: English
• Published: Elsevier 2014-01-01
• Series: Molecular Therapy: Nucleic Acids
• Subjects: CCR5 gene editing; HIV-1 therapy; humanized mice; non-integrating lentivirus; resting CD4+ T cells; zinc finger nucleases
• Online Access: http://www.sciencedirect.com/science/article/pii/S2162253116303353

CCR5 disruption by zinc finger nucleases (ZFNs) is a promising method for HIV-1 gene therapy. However, successful clinical translation of this strategy necessitates the development of a safe and effective method for delivery into relevant cells. We used non-integrating lentivirus (NILV) for transient expression of ZFNs and pseudotyped the virus with HIV-envelope for targeted delivery to CD4+ T cells. Both activated and resting primary CD4+ T cells transduced with CCR5-ZFNs NILV showed resistance to HIV-1 infection in vitro. Furthermore, NILV transduced resting CD4+ T cells from HIV-1 seronegative individuals were resistant to HIV-1 challenge when reconstituted into NOD-scid IL2rγc null (NSG) mice. Likewise, endogenous virus replication was suppressed in NSG mice reconstituted with CCR5-ZFN–transduced resting CD4+ T cells from treatment naïve as well as ART-treated HIV-1 seropositive patients. Taken together, NILV pseudotyped with HIV envelope provides a simple and clinically viable strategy for HIV-1 gene therapy.