Multiplex base editing to protect from CD33 directed drugs for immune and gene therapy

Abstract The selection of genetically engineered immune or hematopoietic cells in vivo after gene editing remains a clinical problem and requires a method to spare on-target toxicity to normal cells. Here, we develop a base editing approach exploiting a naturally occurring CD33 single nucleotide pol...

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Main Authors: Florence Borot, Olivier Humbert, Jeffrey T. Ehmsen, Emily Fields, Sajeev Kohli, Stefan Radtke, Kyle Swing, Dnyanada Pande, Mark R. Enstrom, George S. Laszlo, Thiyagaraj Mayuranathan, Abdullah Mahmood Ali, Mitchell J. Weiss, Jonathan S. Yen, Gregory A. Newby, Roland B. Walter, David R. Liu, Siddhartha Mukherjee, Hans-Peter Kiem
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-59713-2
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Summary:Abstract The selection of genetically engineered immune or hematopoietic cells in vivo after gene editing remains a clinical problem and requires a method to spare on-target toxicity to normal cells. Here, we develop a base editing approach exploiting a naturally occurring CD33 single nucleotide polymorphism leading to removal of full-length CD33 surface expression on edited cells. CD33 editing in human and nonhuman primate hematopoietic stem and progenitor cells protects myeloid progeny from CD33-targeted therapeutics without affecting normal hematopoiesis in vivo, thus demonstrating potential for improved immunotherapies with reduced off-leukemia toxicity. For broader application to gene therapies, we demonstrate highly efficient (>70%) multiplexed adenine base editing of the CD33 and gamma globin genes, resulting in long-term persistence of dual gene-edited cells with HbF reactivation in nonhuman primates. Using the CD33 antibody-drug conjugate Gemtuzumab Ozogamicin, we show resistance of engrafted, multiplex edited human cells in vivo, and a 2-fold enrichment for edited cells in vitro. Together, our results highlight the potential of adenine base editors for improved immune and gene therapies.
ISSN:2041-1723