Efficient clinical scale gene modification via zinc finger nuclease-targeted disruption of the HIV co-receptor CCR5

• Published in: Human gene therapy Vol. 24; no. 3; p. 245
• Main Authors: Maier, Dawn A, Brennan, Andrea L, Jiang, Shuguang, Binder-Scholl, Gwendolyn K, Lee, Gary, Plesa, Gabriela, Zheng, Zhaohui, Cotte, Julio, Carpenito, Carmine, Wood, Travis, Spratt, S Kaye, Ando, Dale, Gregory, Philip, Holmes, Michael C, Perez, Elena E, Riley, James L, Carroll, Richard G, June, Carl H, Levine, Bruce L
• Format: Journal Article
• Language: English
• Published: United States 01.03.2013
• Subjects: Adenoviruses, Human - genetics; Adoptive Transfer; Animals; CD28 Antigens - immunology; CD3 Complex - immunology; DNA Restriction Enzymes - genetics; DNA Restriction Enzymes - metabolism; Female; Genetic Vectors - administration & dosage; Genetic Vectors - adverse effects; Genetic Vectors - genetics; Genetic Vectors - standards; HIV Infections - genetics; HIV Infections - immunology; HIV Infections - therapy; Humans; Lymphocyte Activation - immunology; Male; Mice; Phenotype; Receptors, CCR5 - genetics; Receptors, CCR5 - immunology; T-Lymphocytes - immunology; T-Lymphocytes - metabolism; Transduction, Genetic - methods; Transduction, Genetic - standards; Transplantation, Heterologous; Zinc Fingers - genetics
• ISSN: 1557-7422
• DOI: 10.1089/hum.2012.172

Since HIV requires CD4 and a co-receptor, most commonly C-C chemokine receptor 5 (CCR5), for cellular entry, targeting CCR5 expression is an attractive approach for therapy of HIV infection. Treatment of CD4(+) T cells with zinc-finger protein nucleases (ZFNs) specifically disrupting chemokine receptor CCR5 coding sequences induces resistance to HIV infection in vitro and in vivo. A chimeric Ad5/F35 adenoviral vector encoding CCR5-ZFNs permitted efficient delivery and transient expression following anti-CD3/anti-CD28 costimulation of T lymphocytes. We present data showing CD3/CD28 costimulation substantially improved transduction efficiency over reported methods for Ad5/F35 transduction of T lymphocytes. Modifications to the laboratory scale process, incorporating clinically compatible reagents and methods, resulted in a robust ex vivo manufacturing process capable of generating >10(10) CCR5 gene-edited CD4+ T cells from healthy and HIV+ donors. CD4+ T-cell phenotype, cytokine production, and repertoire were comparable between ZFN-modified and control cells. Following consultation with regulatory authorities, we conducted in vivo toxicity studies that showed no detectable ZFN-specific toxicity or T-cell transformation. Based on these findings, we initiated a clinical trial testing the safety and feasibility of CCR5 gene-edited CD4+ T-cell transfer in study subjects with HIV-1 infection.