Therapeutic base editing of human hematopoietic stem cells
- 16 March 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Medicine
- Vol. 26 (4), 535-541
- https://doi.org/10.1038/s41591-020-0790-y
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.Keywords
Funding Information
- U.S. Department of Health & Human Services | NIH | National Heart, Lung, and Blood Institute (P01HL053749)
- U.S. Department of Health & Human Services | NIH | National Institute of Allergy and Infectious Diseases (R01AI117839)
- U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (R01GM115911, R35 GM118158)
- U.S. Department of Health & Human Services | NIH | National Human Genome Research Institute (R00HG008399, RM1 HG009490)
This publication has 33 references indexed in Scilit:
- A Novel Promoter Mutation (HBB: c.-75G>T) Was Identified as a Cause of β+-ThalassemiaHemoglobin, 2015
- Nonirradiated NOD,B6.SCID Il2rγ−/− KitW41/W41 (NBSGW) Mice Support Multilineage Engraftment of Human Hematopoietic CellsStem Cell Reports, 2015
- Targeted genome editing in human repopulating haematopoietic stem cellsNature, 2014
- Gene Editing ofCCR5in Autologous CD4 T Cells of Persons Infected with HIVThe New England Journal of Medicine, 2014
- Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleasesBioinformatics, 2014
- An Erythroid Enhancer of BCL11A Subject to Genetic Variation Determines Fetal Hemoglobin LevelScience, 2013
- Hematopoietic Stem Cell Quiescence Promotes Error-Prone DNA Repair and MutagenesisCell Stem Cell, 2010
- Human hematopoietic stem/progenitor cells modified by zinc-finger nucleases targeted to CCR5 control HIV-1 in vivoNature Biotechnology, 2010
- Three New β-Globin Gene Promoter Mutations Identified Through Newborn ScreeningHemoglobin, 2007
- Evolutionarily conserved elements in vertebrate, insect, worm, and yeast genomesGenome Research, 2005