Correction of the sickle cell disease mutation in human hematopoietic stem/progenitor cells

MD Hoban, GJ Cost, MC Mendel… - Blood, The Journal …, 2015 - ashpublications.org
MD Hoban, GJ Cost, MC Mendel, Z Romero, ML Kaufman, AV Joglekar, M Ho, D Lumaquin
Blood, The Journal of the American Society of Hematology, 2015ashpublications.org
Sickle cell disease (SCD) is characterized by a single point mutation in the seventh codon of
the β-globin gene. Site-specific correction of the sickle mutation in hematopoietic stem cells
would allow for permanent production of normal red blood cells. Using zinc-finger nucleases
(ZFNs) designed to flank the sickle mutation, we demonstrate efficient targeted cleavage at
the β-globin locus with minimal off-target modification. By codelivering a homologous donor
template (either an integrase-defective lentiviral vector or a DNA oligonucleotide), high …
Abstract
Sickle cell disease (SCD) is characterized by a single point mutation in the seventh codon of the β-globin gene. Site-specific correction of the sickle mutation in hematopoietic stem cells would allow for permanent production of normal red blood cells. Using zinc-finger nucleases (ZFNs) designed to flank the sickle mutation, we demonstrate efficient targeted cleavage at the β-globin locus with minimal off-target modification. By codelivering a homologous donor template (either an integrase-defective lentiviral vector or a DNA oligonucleotide), high levels of gene modification were achieved in CD34+ hematopoietic stem and progenitor cells. Modified cells maintained their ability to engraft NOD/SCID/IL2rγnull mice and to produce cells from multiple lineages, although with a reduction in the modification levels relative to the in vitro samples. Importantly, ZFN-driven gene correction in CD34+ cells from the bone marrow of patients with SCD resulted in the production of wild-type hemoglobin tetramers.
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