对于药物耐受性增强的增殖持久性存在着很多未探明的驱动因素。组蛋白H3赖氨酸-4三甲基化（H3K4me3），作为一种活性组蛋白标记，可能使缓慢周期的耐药持久性存活细胞（DTP）恢复增殖能力。本研究旨在确定H3K4me3在转录活跃位点上的存在，以鉴定DTPP的关键调控因子。采用适应性肿瘤耐药模型，通过H3K4me3 ChIP-Seq数据引导识别DTPP的顶级转录因子结合基序。这些结果提示，OGT（O-β-N-乙酰葡萄糖胺转移酶）可能参与其中，这得到了代谢组学分析和生化实验的证实。通过体内外研究OGT对DTPP和适应性耐药的影响。H3K4me3重塑普遍存在于CpG岛区域，并且与O-GlcNAc标记的染色质相关的DNA结合基序相互关联。因此，我们观察到在长期处理的癌细胞中OGT、O-GlcNAc及其结合伴侣TET1的上调。抑制OGT导致H3K4me3的丧失和抗药性相关基因的下调。OGT的遗传敲除防止了在体内模型中获得的抗药性。在OGT上游，我们确定了AMPK作为一个可操作的靶点。乙酰水杨酸通过激活AMPK下调了OGT，并对延迟获得性耐药性产生了类似的效果。我们的发现揭示了一种适应性药物耐药性的基本机制，该机制控制着癌细胞向获得性药物耐药性的重编程，这一过程可被利用以提高治疗响应持续时间和患者预后。版权所有 © 2023 作者。由Elsevier Ltd.出版。保留所有权利。

Drivers of the drug tolerant proliferative persister (DTPP) state have not been well investigated. Histone H3 lysine-4 trimethylation (H3K4me3), an active histone mark, might enable slow cycling drug tolerant persisters (DTP) to regain proliferative capacity. This study aimed to determine H3K4me3 transcriptionally active sites identifying a key regulator of DTPPs.Deploying a model of adaptive cancer drug tolerance, H3K4me3 ChIP-Seq data of DTPPs guided identification of top transcription factor binding motifs. These suggested involvement of O-linked N-acetylglucosamine transferase (OGT), which was confirmed by metabolomics analysis and biochemical assays. OGT impact on DTPPs and adaptive resistance was explored in vitro and in vivo.H3K4me3 remodeling was widespread in CPG island regions and DNA binding motifs associated with O-GlcNAc marked chromatin. Accordingly, we observed an upregulation of OGT, O-GlcNAc and its binding partner TET1 in chronically treated cancer cells. Inhibition of OGT led to loss of H3K4me3 and downregulation of genes contributing to drug resistance. Genetic ablation of OGT prevented acquired drug resistance in in vivo models. Upstream of OGT, we identified AMPK as an actionable target. AMPK activation by acetyl salicylic acid downregulated OGT with similar effects on delaying acquired resistance.Our findings uncover a fundamental mechanism of adaptive drug resistance that governs cancer cell reprogramming towards acquired drug resistance, a process that can be exploited to improve response duration and patient outcomes.Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.