腺苷脱氨酶作用于RNA1（ADAR1）在应激反应期间通过防止逆转录病毒整合和逆转底物保护基因组完整性。然而，炎症微环境诱导ADAR1p110至p150剪接异构体切换导致20种恶性肿瘤中的癌症干细胞（CSC）的生成和治疗抵抗性。之前，预测和预防ADAR1p150介导的恶性RNA编辑代表了一个重大的挑战。因此，我们开发了基于慢病毒的ADAR1和剪接报告基因，以非侵入性检测剪接介导的ADAR1腺苷-肌苷（A-to-I）RNA编辑激活；定量的ADAR1p150细胞内流式细胞术；一种选择性小分子抑制剪接介导的ADAR1激活的药物Rebecsinib，在剂量可以节省正常造血干细胞（HSPCs）的情况下抑制白血病干细胞（LSC）的自我更新并延长人类化LSC小鼠模型的生存期；和预先IND研究显示良好的Rebecsinib毒代动力学和药效学（TK/PD）特性。这些结果为将Rebecsinib作为临床ADAR1p150拮抗剂的发展奠定了基础，旨在消除恶性微环境驱动的LSC生成。版权所有©2023年作者。由Elsevier Inc.出版。保留所有权利。

Adenosine deaminase acting on RNA1 (ADAR1) preserves genomic integrity by preventing retroviral integration and retrotransposition during stress responses. However, inflammatory-microenvironment-induced ADAR1p110 to p150 splice isoform switching drives cancer stem cell (CSC) generation and therapeutic resistance in 20 malignancies. Previously, predicting and preventing ADAR1p150-mediated malignant RNA editing represented a significant challenge. Thus, we developed lentiviral ADAR1 and splicing reporters for non-invasive detection of splicing-mediated ADAR1 adenosine-to-inosine (A-to-I) RNA editing activation; a quantitative ADAR1p150 intracellular flow cytometric assay; a selective small-molecule inhibitor of splicing-mediated ADAR1 activation, Rebecsinib, which inhibits leukemia stem cell (LSC) self-renewal and prolongs humanized LSC mouse model survival at doses that spare normal hematopoietic stem and progenitor cells (HSPCs); and pre-IND studies showing favorable Rebecsinib toxicokinetic and pharmacodynamic (TK/PD) properties. Together, these results lay the foundation for developing Rebecsinib as a clinical ADAR1p150 antagonist aimed at obviating malignant microenvironment-driven LSC generation.Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.