Efficient clinical scale gene modification via zinc finger nuclease-targeted disruption of the HIV co-receptor CCR5
Open accessAbstract
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.
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The same group scaled up the process and derived 10^10 gene-modified CD4+ T cells using AD5/F35 adenoviral vector for transduction and anti-CD3/anti-CD28 stimulation for expansion.
“The same group has recently scaled up the process and was able to derive 1010 gene modified CD4+ T cells by using AD5/F35 adenoviral vector for transduction and anti-CD3/anti-CD28 stimulation for expansion of CD4+ T cells [41].”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [41]
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- Authors
- Maier, D. A., Brennan, A. L., Jiang, S., Binder-Scholl, G. K., Lee, G., Plesa, G., Zheng, Z., Cotte, J., Carpenito, C., Wood, T., Spratt, S. K., Ando, D., Gregory, P., Holmes, M. C., Perez, E. E., Riley, J. L., Carroll, R. G., June, C. H., & Levine, B. L.
- Affiliations
- University of Pennsylvania (US)
- Sangamo BioSciences (United States) (US)
- University of Pennsylvania Perelman School of Medicine
- Corresponding author
- Levine, Bruce L.
- ORCID
- Venue
- Human Gene Therapy
- Publisher
- SAGE Publications
- Published
- 2013-01-29
- Volume
- 24
- Issue
- 3
- Pages
- 245-258
- ISSN
- 1043-0342 (ISSN-L)
- 1557-7422
- PMC
- PMC3609630
- OpenAlex
- W2090994613
- Cited by
- 126
- References
- 78
- Citations by year
- 2026: 2 · 2025: 3 · 2024: 5 · 2023: 5 · 2022: 2 · 2021: 7 · 2020: 4 · 2019: 4 · 2018: 13 · 2017: 6 · 2016: 16 · 2015: 21 · 2014: 25 · 2013: 13
- FWCI
- 16.59
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Human Gene Therapy (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Institute of Allergy and Infectious Diseases (R01AI104400, U19AI082628, U19AI066290)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Immunology and Microbiology › Virology › HIV Research and Treatment
- Keywords
- HIV-1 infection, zinc finger nucleases, HIV co-receptor, CD4+ T cells, ex vivo T cell engineering, adenoviral vector, anti-CD3/CD28 stimulation, HIV drug resistance, T-cell transformation, clinical trial safety, HIV gene therapy
- MeSH terms
- Adenoviruses, Human, Animals, DNA Restriction Enzymes, Female, Genetic Vectors, Humans, Lymphocyte Activation, Male, Phenotype, T-Lymphocytes, Transduction, Genetic, Transplantation, Heterologous, HIV Infections, Zinc Fingers, CD3 Complex, CD28 Antigens, Adoptive Transfer, Receptors, CCR5, Mice
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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