1. Genomic Editing of the HIV-1 Coreceptor CCR5 in Adult Hematopoietic Stem and Progenitor Cells Using Zinc Finger Nucleases.
- Author
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Li, Lijing, Krymskaya, Ludmila, Wang, Jianbin, Henley, Jill, Rao, Anitha, Cao, Lan-Feng, Tran, Chy-Anh, Torres-Coronado, Monica, Gardner, Agnes, Gonzalez, Nancy, Kim, Kenneth, Liu, Pei-Qi, Hofer, Ursula, Lopez, Evan, Gregory, Philip D, Liu, Qing, Holmes, Michael C, Cannon, Paula M, Zaia, John A, and DiGiusto, David L
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AIDS treatment , *HIV , *STEM cell research , *HEMATOPOIETIC stem cells , *PROTEIN kinases - 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 (CCR5Δ32/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 CCR5Δ32/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. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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