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In vivo base editing by a single i.v. vector injection for treatment of hemoglobinopathies
Chang Li, … , Evangelia Yannaki, André Lieber
Chang Li, … , Evangelia Yannaki, André Lieber
Published August 25, 2022
Citation Information: JCI Insight. 2022;7(19):e162939. https://doi.org/10.1172/jci.insight.162939.
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Research Article Hematology Stem cells

In vivo base editing by a single i.v. vector injection for treatment of hemoglobinopathies

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Abstract

Individuals with β-thalassemia or sickle cell disease and hereditary persistence of fetal hemoglobin (HPFH) possessing 30% fetal hemoglobin (HbF) appear to be symptom free. Here, we used a nonintegrating HDAd5/35++ vector expressing a highly efficient and accurate version of an adenine base editor (ABE8e) to install, in vivo, a –113 A>G HPFH mutation in the γ-globin promoters in healthy CD46/β-YAC mice carrying the human β-globin locus. Our in vivo hematopoietic stem cell (HSC) editing/selection strategy involves only s.c. and i.v. injections and does not require myeloablation and HSC transplantation. In vivo HSC base editing in CD46/β-YAC mice resulted in > 60% –113 A>G conversion, with 30% γ-globin of β-globin expressed in 70% of erythrocytes. Importantly, no off-target editing at sites predicted by CIRCLE-Seq or in silico was detected. Furthermore, no critical alterations in the transcriptome of in vivo edited mice were found by RNA-Seq. In vitro, in HSCs from β-thalassemia and patients with sickle cell disease, transduction with the base editor vector mediated efficient –113 A>G conversion and reactivation of γ-globin expression with subsequent phenotypic correction of erythroid cells. Because our in vivo base editing strategy is safe and technically simple, it has the potential for clinical application in developing countries where hemoglobinopathies are prevalent.

Authors

Chang Li, Aphrodite Georgakopoulou, Gregory A. Newby, Kelcee A. Everette, Evangelos Nizamis, Kiriaki Paschoudi, Efthymia Vlachaki, Sucheol Gil, Anna K. Anderson, Theodore Koob, Lishan Huang, Hongjie Wang, Hans-Peter Kiem, David R. Liu, Evangelia Yannaki, André Lieber

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Figure 11

Differentially expressed genes after in vivo base editing HDAd-EF1α.ABE8e.

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Differentially expressed genes after in vivo base editing HDAd-EF1α.ABE8...
RNA-Seq analysis was performed with BM MNCs collected before treatment and at week 16 after in vivo HSC transduction/selection with HDAd-EF1α.ABE8e (n = 3 females, age 8–12 weeks). The genome of CD46/β-YAC mice contains the WT 248 kb β-globin locus (β-YAC) including the LCR, γ- and β-globin genes. Shown are genes with altered mRNA expression based on their adjusted P value after annotation with the human genome. (A) Altered mRNA expression after annotation with a reference human genome. Statistically significant changes (P < 0.01) are highlighted in red (upregulated). (B) Volcano plot of mRNA data after annotation with the mouse reference genome are shown. Statistically significant changes (P < 0.01) are highlighted in red (upregulated) or blue (downregulated). Statistical analyses were performed using 2-way ANOVA.

Copyright © 2023 American Society for Clinical Investigation
ISSN 2379-3708

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