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Genomic Editing of the HIV-1 Coreceptor CCR5 in Adult Hematopoietic Stem and Progenitor Cells Using Zinc Finger Nucleases.
- Source :
-
Molecular Therapy . Jun2013, Vol. 21 Issue 6, p1259-1269. 11p. - Publication Year :
- 2013
-
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]
- Subjects :
- *AIDS treatment
*HIV
*STEM cell research
*HEMATOPOIETIC stem cells
*PROTEIN kinases
Subjects
Details
- Language :
- English
- ISSN :
- 15250016
- Volume :
- 21
- Issue :
- 6
- Database :
- Academic Search Index
- Journal :
- Molecular Therapy
- Publication Type :
- Academic Journal
- Accession number :
- 87926346
- Full Text :
- https://doi.org/10.1038/mt.2013.65