NOTCH-Delta-HES信号级联由于其在不同细胞环境中的双重肿瘤抑制和致癌作用而被认为是一把双刃剑。在 T 细胞中，它通过促进分化来支持白血病发生，而在 B 细胞中，它通过抑制早期分化/诱导生长停滞从而导致细胞凋亡来控制白血病发生。本研究旨在评估 NOTCH 家族的这种双向行为是否可以用作急性淋巴细胞白血病 (ALL) 的诊断生物标志物或亚型分类器。在此研究中，使用定量 PCR (qPCR) 分析了儿科 ALL 患者的骨髓 (BM) 活检和血浆 (BP) 中七个 NOTCH 级联基因的表达。此外，通过甲基化特异性 qMSP 评估差异表达基因 (DEG) 的启动子 DNA 甲基化状态，并通过亚硫酸氢盐扩增子测序进行验证。尽管在所有患者中均观察到 JAG1、DLL1、HES-2、HES-4 和 HES-5 的高甲基化，但发现 NOTCH3 特别是在 Pre-B ALL 病例中高度甲基化，而 DLL4 在 Pre-T ALL 病例中高度甲基化。异常的 DNA 甲基化与下调的基因表达密切相关，如在“随访/治疗后”受试者中观察到的，基因表达在完全缓解阶段恢复。亚型特异性 ROC 曲线分析和 Kaplan-Meier 生存分析预测了该面板的临床适用诊断和预后潜力。此外，逻辑回归模型（Pre-B vs Pre-T ALL）被发现是最佳拟合模型（McFadden 的 R2 = 0.28，F1 测量 = 0.99）。无论是在 BM 抽吸物还是血浆中进行分析，NOTCH 表观遗传特征都显示出可比较的结果 (p< 0.001)，这表明 NOTCH-Delta-HES 级联作为亚型分类器在 ALL 微创诊断中的潜力。© 2023。作者获得 Springer-Verlag GmbH（德国 Springer Nature 旗下公司）的独家许可。

The NOTCH-Delta-HES signaling cascade is regarded as a double-edged sword owing to its dual tumor-suppressor and oncogenic roles, in different cellular environments. In the T-cells, it supports leukemogenesis by promoting differentiation while in B-cells, it controls leukemogenesis by inhibiting early differentiation/inducing growth arrest in the lead to apoptosis. The present study was undertaken to assess if this bi-faceted behavior of NOTCH family can be exploited as a diagnostic biomarker or subtype classifier of acute lymphoblastic leukemia (ALL). In this pursuit, expression of seven NOTCH cascade genes was analyzed in bone marrow (BM) biopsy and blood plasma (BP) of pediatric ALL patients using quantitative PCR (qPCR). Further, promoter DNA methylation status of the differentially expressed genes (DEGs) was assessed by methylation-specific qMSP and validated through bisulphite amplicon sequencing. Whereas hypermethylation of JAG1, DLL1, and HES-2, HES-4, and HES-5 was observed in all patients, NOTCH3 was found hypermethylated specifically in Pre-B ALL cases while DLL4 in Pre-T ALL cases. Aberrant DNA methylation strongly correlated with downregulated gene expression, which restored at complete remission stage as observed in "follow-up/post-treatment" subjects. The subtype-specific ROC curve analysis and Kaplan-Meier survival analysis predicted a clinically applicable diagnostic and prognostic potential of the panel. Moreover, the logistic regression model (Pre-B vs Pre-T ALL) was found to be the best-fitted model (McFadden's R2 = 0.28, F1 measure = 0.99). Whether analyzed in BM-aspirates or blood plasma, the NOTCH epigenetic signatures displayed comparable results (p < 0.001), advocating the potential of NOTCH-Delta-HES cascade, as a subtype classifier, in minimally invasive diagnosis of ALL.© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.