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

Abstract

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.

• Authors: Dawn A Maier, Andrea L Brennan, Shuguang Jiang, Gwendolyn K Binder-Scholl, Gary Lee, Gabriela Plesa, Zhaohui Zheng, Julio Cotte, Carmine Carpenito, Travis Wood, S Kaye Spratt, Dale Ando, Philip Gregory, Michael C Holmes, Elena E Perez, James L Riley, Richard G Carroll, Carl H June, Bruce L Levine
• Journal: Human gene therapy (Hum Gene Ther) Vol. 24 Issue 3 Pg. 245-58 (Mar 2013) ISSN: 1557-7422 [Electronic] United States
• PMID: 23360514 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• CD28 Antigens
• CD3 Complex
• Receptors, CCR5
• DNA Restriction Enzymes

Topics

• Adenoviruses, Human (genetics)
• Adoptive Transfer
• Animals
• CD28 Antigens (immunology)
• CD3 Complex (immunology)
• DNA Restriction Enzymes (genetics, metabolism)
• Female
• Genetic Vectors (administration & dosage, adverse effects, genetics, standards)
• HIV Infections (genetics, immunology, therapy)
• Humans
• Lymphocyte Activation (immunology)
• Male
• Mice
• Phenotype
• Receptors, CCR5 (genetics, immunology)
• T-Lymphocytes (immunology, metabolism)
• Transduction, Genetic (methods, standards)
• Transplantation, Heterologous
• Zinc Fingers (genetics)