Go to The Journal of Clinical Investigation
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising
  • Job board
  • Contact
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Immunology
    • Metabolism
    • Nephrology
    • Oncology
    • Pulmonology
    • All ...
  • Videos
  • Collections
    • Resource and Technical Advances
    • Clinical Medicine
    • Reviews
    • Editorials
    • Perspectives
    • Top read articles
  • JCI This Month
    • Current issue
    • Past issues

  • Current issue
  • Past issues
  • Specialties
  • In-Press Preview
  • Editorials
  • Viewpoint
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising
  • Job board
  • Contact
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.
View: Text | PDF
Research Article Hematology Stem cells

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

  • Text
  • PDF
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

×

Figure 4

Cytospins from cultures at different time points of ED.

Options: View larger image (or click on image) Download as PowerPoint
Cytospins from cultures at different time points of ED.
(A) Representati...
(A) Representative samples from Thal CD34+ cells either untransduced or transduced with HDAd-EF1α.ABE8e and O6-BG/BCNU selected. Left panel: cytospins stained with Giemsa/May-Grünwald (see Supplemental Methods). The arrows indicate cells at different stages of erythroid differentiation: proerythroblasts (red) → basophilic erythroblasts (green) → polychromatic erythroblasts (blue) → orthochromatic erythroblasts (orange) → maturing orthochromatic erythroblasts (yellow) → reticulocytes (black)/pyrenocytes (gray). Scale bar: 25 μm. Right panel: quantification of erythroid progenitors. Five random fields were counted by 2 scientists. Pro, proerythroblasts; Baso, basophilic erythroblasts; Poly, polychromatic erythroblasts; Ortho, orthochromatic erythroblasts; mat; Ortho, maturing orthochromatic erythroblasts; Retics, reticulocytes; Pyrcs, pyrenocytes. (B) Cytospins from differentiated SCD CD34+ cells at days 11, 14, and 18 of ED, untransduced and HDAd-EF1α.ABE8e –transduced without and with selection. Statistical analyses were performed using 2-way ANOVA. *P < 0.05.

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

Sign up for email alerts