目的：神经母细胞瘤（NB）是一种周围神经系统的儿科恶性肿瘤，其特征是表观遗传和转录（EP-TF）异常。本研究旨在利用 CRISPR-Cas9 敲除筛选开发 NB 的 EP-TF 临床预后模型。结果：使用 CRISPR-Cas9 筛选和公共 NB 数据集进行体外和体内综合分析，以确定 35 个 EP-TF 基因，这些基因在 NB 中表现出最高表达，并且高度依赖于癌症生存能力。经过单变量分析，这 35 个基因中的 27 个被纳入最小绝对收缩和选择算子筛选中。我们在不同的 NB 队列中建立了一个预后 EP-TF 模型并进行了生物学验证，该模型包含 RUVBL1、LARP7、GTF3C4、THAP10、SUPT16H、TIGD1、SUV39H2、TAF1A、SMAD9 和 FEM1B。 MYCN 是 EP-TF 基因的潜在上游调节因子。高风险亚型表现出与恶性细胞周期、MYCN 相关信号传导和染色质重塑相关的特征，所有这些特征都与不良预后和免疫抑制相关。 MEK 抑制剂已成为针对 NB 中大多数 EP-TF 风险基因的有前景的治疗药物。结论：我们的新型预后模型在预测和评估 NB 患者的总体生存率方面显示出巨大的潜力，为治疗目标提供见解。版权所有 © 2024 张、莫、石、熊、艾尔肯、陈、唐、赵、吕和谭。

Objectives: Neuroblastoma (NB), a pediatric malignancy of the peripheral nervous system, is characterized by epigenetic and transcriptional (EP-TF) anomalies. This study aimed to develop an EP-TF clinical prognostic model for NB using CRISPR-Cas9 knockout screening. Results: An integrative analysis was conducted using CRISPR-Cas9 screening in vitro and in vivo with public NB datasets to identify 35 EP-TF genes that exhibited the highest expression in NB and were highly dependent on cancer viability. After univariate analysis, 27 of these 35 genes were included in the least absolute shrinkage and selection operator screen. We established and biologically validated a prognostic EP-TF model encompassing RUVBL1, LARP7, GTF3C4, THAP10, SUPT16H, TIGD1, SUV39H2, TAF1A, SMAD9, and FEM1B across diverse NB cohorts. MYCN serves a potential upstream regulator of EP-TF genes. The high-risk subtype exhibited traits associated with the malignant cell cycle, MYCN-linked signaling and chromatin remodeling, all of which are correlated with poor prognosis and immunosuppression. MEK inhibitors have emerged as promising therapeutic agents for targeting most EP-TF risk genes in NB. Conclusion: Our novel prognostic model shows significant potential for predicting and evaluating the overall survival of NB patients, offering insights into therapeutic targets.Copyright © 2024 Zhang, Mo, Shi, Xiong, Aierken, Chen, Tang, Zhao, Lv and Tan.