Cited by 126Virus-based gene therapy researchCRISPR and Genetic EngineeringHIV Research and Treatment

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

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Maier, Dawn A., Brennan, Andrea L., Jiang, Shuguang, Binder-Scholl, Gwendolyn K., Lee, Gary, Plesa, Gabriela, Zheng, Zhaohui, Cotte, Julio +11 more·2013·Human Gene Therapy

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.

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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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  1. Hurez V. Dzialo-Hatton R. Oliver J., et al. Efficient adenovirus-mediated gene transfer into primary T cells and thymocytes in a new coxsackie/adenovirus receptor transgenic model. BMC. Immunol. 2002;3:4. doi: 10.1186/1471-2172-3-4. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI

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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.
Venue
Human Gene Therapy
Published
2013-01-29
Volume
24
Issue
3
Pages
245-258
ISSN
  • 1043-0342 (ISSN-L)
  • 1557-7422
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
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

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