Title |
Genomic Editing of the HIV-1 Coreceptor CCR5 in Adult Hematopoietic Stem and Progenitor Cells Using Zinc Finger Nucleases
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Published in |
Molecular Therapy, April 2013
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DOI | 10.1038/mt.2013.65 |
Pubmed ID | |
Authors |
Lijing Li, Ludmila Krymskaya, Jianbin Wang, Jill Henley, Anitha Rao, Lan-Feng Cao, Chy-Anh Tran, Monica Torres-Coronado, Agnes Gardner, Nancy Gonzalez, Kenneth Kim, Pei-Qi Liu, Ursula Hofer, Evan Lopez, Philip D Gregory, Qing Liu, Michael C Holmes, Paula M Cannon, John A Zaia, David L DiGiusto |
Abstract |
The HIV-1 coreceptor CCR5 is a validated target for HIV/AIDS therapy. The apparent elimination of HIV-1 in a patient treated with an allogeneic stem cell transplant homozygous for a naturally occurring CCR5 deletion mutation (CCR5(Δ32/Δ32)) supports the concept that a single dose of HIV-resistant hematopoietic stem cells can provide disease protection. Given the low frequency of naturally occurring CCR5(Δ32/Δ32) donors, we reasoned that engineered autologous CD34(+) hematopoietic stem/progenitor cells (HSPCs) could be used for AIDS therapy. We evaluated disruption of CCR5 gene expression in HSPCs isolated from granulocyte colony-stimulating factor (CSF)-mobilized adult blood using a recombinant adenoviral vector encoding a CCR5-specific pair of zinc finger nucleases (CCR5-ZFN). Our results demonstrate that CCR5-ZFN RNA and protein expression from the adenoviral vector is enhanced by pretreatment of HSPC with protein kinase C (PKC) activators resulting in >25% CCR5 gene disruption and that activation of the mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling pathway is responsible for this activity. Importantly, using an optimized dose of PKC activator and adenoviral vector we could generate CCR5-modified HSPCs which engraft in a humanized mouse model (albeit at a reduced level) and support multilineage differentiation in vitro and in vivo. Together, these data establish the basis for improved approaches exploiting adenoviral vector delivery in the modification of HSPCs. |
X Demographics
Geographical breakdown
Country | Count | As % |
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United States | 5 | 83% |
Unknown | 1 | 17% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 5 | 83% |
Scientists | 1 | 17% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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United States | 5 | 2% |
Sweden | 1 | <1% |
Germany | 1 | <1% |
Spain | 1 | <1% |
India | 1 | <1% |
Unknown | 195 | 96% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 45 | 22% |
Researcher | 43 | 21% |
Student > Bachelor | 32 | 16% |
Student > Master | 26 | 13% |
Student > Doctoral Student | 10 | 5% |
Other | 24 | 12% |
Unknown | 24 | 12% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 76 | 37% |
Biochemistry, Genetics and Molecular Biology | 46 | 23% |
Medicine and Dentistry | 28 | 14% |
Immunology and Microbiology | 9 | 4% |
Engineering | 7 | 3% |
Other | 13 | 6% |
Unknown | 25 | 12% |