Abstract
Lentivector gene therapy for X-linked chronic granulomatous disease (X-CGD) has proven to be a viable approach, but random vector integration and subnormal protein production from exogenous promoters in transduced cells remain concerning for long-term safety and efficacy. A previous genome editing-based approach using Streptococcus pyogenes Cas9 mRNA and an oligodeoxynucleotide donor to repair genetic mutations showed the capability to restore physiological protein expression but lacked sufficient efficiency in quiescent CD34+ hematopoietic cells for clinical translation. Here, we report that transient inhibition of p53-binding protein 1 (53BP1) significantly increased (2.3-fold) long-term homology-directed repair to achieve highly efficient (80% gp91phox+ cells compared with healthy donor control subjects) long-term correction of X-CGD CD34+ cells.
Publication types
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Research Support, N.I.H., Intramural
MeSH terms
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Animals
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Bacterial Proteins
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Caspase 9
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Cells, Cultured
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DNA Repair* / genetics
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Dependovirus / genetics
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Exons / genetics
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Gene Editing / methods*
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Genetic Therapy / methods*
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Genetic Vectors / genetics
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Genetic Vectors / therapeutic use
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Granulomatous Disease, Chronic / genetics
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Granulomatous Disease, Chronic / therapy*
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Hematopoietic Stem Cell Transplantation*
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Hematopoietic Stem Cells / enzymology
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Heterografts
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Humans
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Male
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Mice
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Mice, Inbred NOD
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Mice, SCID
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NADPH Oxidase 2 / deficiency
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NADPH Oxidase 2 / genetics*
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Phagocytes / metabolism
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RNA, Messenger / genetics
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Reactive Oxygen Species
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Ribonucleoproteins / genetics
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Sequence Deletion
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Streptococcus pyogenes / enzymology
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Tumor Suppressor p53-Binding Protein 1 / antagonists & inhibitors*
Substances
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Bacterial Proteins
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RNA, Messenger
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Reactive Oxygen Species
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Ribonucleoproteins
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TP53BP1 protein, human
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Tumor Suppressor p53-Binding Protein 1
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CYBB protein, human
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NADPH Oxidase 2
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Caspase 9