Therapeutic Base Editing of Human Hematopoietic Stem Cells

Overview

Journal Nat Med

Specialties General Medicine
Molecular Biology

Date 2020 Apr 15

PMID 32284612

Citations 160

Authors

Jing Zeng

Yuxuan Wu

Chunyan Ren

Jasmine Bonanno

Anne H Shen

Devlin Shea

Jason M Gehrke

Kendell Clement

Kevin Luk

Qiuming Yao

Rachel Kim

Scot A Wolfe

John P Manis

Luca Pinello

J Keith Joung

Daniel E Bauer

Affiliations

Soon will be listed here.

Abstract

Base editing by nucleotide deaminases linked to programmable DNA-binding proteins represents a promising approach to permanently remedy blood disorders, although its application in engrafting hematopoietic stem cells (HSCs) remains unexplored. In this study, we purified A3A (N57Q)-BE3 base editor for ribonucleoprotein (RNP) electroporation of human-peripheral-blood-mobilized CD34 hematopoietic stem and progenitor cells (HSPCs). We observed frequent on-target cytosine base edits at the BCL11A erythroid enhancer at +58 with few indels. Fetal hemoglobin (HbF) induction in erythroid progeny after base editing or nuclease editing was similar. A single therapeutic base edit of the BCL11A enhancer prevented sickling and ameliorated globin chain imbalance in erythroid progeny from sickle cell disease and β-thalassemia patient-derived HSPCs, respectively. Moreover, efficient multiplex editing could be achieved with combined disruption of the BCL11A erythroid enhancer and correction of the HBB -28A>G promoter mutation. Finally, base edits could be produced in multilineage-repopulating self-renewing human HSCs with high frequency as assayed in primary and secondary recipient animals resulting in potent HbF induction in vivo. Together, these results demonstrate the potential of RNP base editing of human HSPCs as a feasible alternative to nuclease editing for HSC-targeted therapeutic genome modification.

Citing Articles

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From bench to bedside: cutting-edge applications of base editing and prime editing in precision medicine.

Xu W, Zhang S, Qin H, Yao K J Transl Med. 2024; 22(1):1133.

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Direct delivery of Cas-embedded cytosine base editors as ribonucleoprotein complexes for efficient and accurate editing of clinically relevant targets.

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Gene editing without ex vivo culture evades genotoxicity in human hematopoietic stem cells.

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