Title | Correction of the sickle cell disease mutation in human hematopoietic stem/progenitor cells. |
Publication Type | Journal Article |
Year of Publication | 2015 |
Authors | Hoban MD, Cost GJ, Mendel MC, Romero Z, Kaufman ML, Joglekar AV, Ho M, Lumaquin D, Gray D, Lill GR, Cooper AR, Urbinati F, Senadheera S, Zhu A, Liu P-Q, Paschon DE, Zhang L, Rebar EJ, Wilber A, Wang X, Gregory PD, Holmes MC, Reik A, Hollis RP, Kohn DB |
Journal | Blood |
Volume | 125 |
Issue | 17 |
Pagination | 2597-604 |
Date Published | 2015 Apr 23 |
ISSN | 1528-0020 |
Keywords | Anemia, Sickle Cell, Animals, Antigens, CD34, Base Sequence, beta-Globins, Bone Marrow Cells, Cells, Cultured, Endodeoxyribonucleases, Fetal Blood, Genetic Loci, Genetic Therapy, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells, Humans, Mice, Mice, Inbred NOD, Mice, SCID, Molecular Sequence Data, Mutation, Zinc Fingers |
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 co-delivering 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/IL2rγ(null) 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. |
DOI | 10.1182/blood-2014-12-615948 |
Alternate Journal | Blood |
PubMed ID | 25733580 |
PubMed Central ID | PMC4408287 |
Grant List | GM007185 / GM / NIGMS NIH HHS / United States 5 T32 AI060567 / AI / NIAID NIH HHS / United States R25 GM055052 / GM / NIGMS NIH HHS / United States T32 GM007185 / GM / NIGMS NIH HHS / United States 2P01 HL073104 / HL / NHLBI NIH HHS / United States T32 AI060567 / AI / NIAID NIH HHS / United States P01 HL073104 / HL / NHLBI NIH HHS / United States |
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