Epitope prime editing shields hematopoietic cells from CD123 immunotherapy for acute myeloid leukemia

Cell Stem Cell. 2024 Sep 26:S1934-5909(24)00317-5. doi: 10.1016/j.stem.2024.09.003. Online ahead of print.

Abstract

Acute myeloid leukemia (AML) is a malignant cancer characterized by abnormal differentiation of hematopoietic stem and progenitor cells (HSPCs). While chimeric antigen receptor T (CAR-T) cell immunotherapies target AML cells, they often induce severe on-target/off-tumor toxicity by attacking normal cells expressing the same antigen. Here, we used base editors (BEs) and a prime editor (PE) to modify the epitope of CD123 on HSPCs, protecting healthy cells from CAR-T-induced cytotoxicity while maintaining their normal function. Although BE effectively edits epitopes, complex bystander products are a concern. To enhance precision, we optimized prime editing, increasing the editing efficiency from 5.9% to 78.9% in HSPCs. Epitope-modified cells were resistant to CAR-T lysis while retaining normal differentiation and function. Furthermore, BE- or PE-edited HSPCs infused into humanized mice endowed myeloid lineages with selective resistance to CAR-T immunotherapy, demonstrating a proof-of-concept strategy for treating relapsed AML.

Keywords: CAR-T immunotherapy; CD123; HSPCs; acute myeloid leukemia; base editing; bystander products; epitope editing; hematopoietic stem and progenitor cell; on-target off-tumor toxicity; prime editing.