超级增强子相关转录因子网络定义了神经母细胞瘤 (NB) 中的细胞身份。这些转录因子的失调通过强制早期发育身份状态来促进 NB 的启动和维持。我们报告，I 类碱性螺旋-环-螺旋 (bHLH) 转录因子 4 (TCF4；也称为 E2-2) 是一种关键的 NB 依赖性基因，它通过与细胞身份特异性 bHLH 的异二聚化对这些身份状态做出显着贡献转录因子。 TCF4 的敲低可显着诱导体外细胞凋亡并抑制体内致瘤性。我们使用全基因组表达谱、TCF4 染色质免疫沉淀测序 (ChIP-seq) 和 TCF4 免疫沉淀-质谱法来确定 TCF4 在 NB 细胞中的作用。我们的结果，连同 NB 中转录因子 T-box 转录因子 TBX2、心脏和神经嵴衍生物表达蛋白 2 (HAND2) 和扭曲相关蛋白 1 (TWIST1) 的最新发现，提出了 TCF4 在调节中的作用。叉头盒蛋白 M1 (FOXM1)/转录因子 E2F 驱动的基因调控网络与 N-myc 原癌基因蛋白 (MYCN)、TBX2 和 TCF4 二聚化伙伴 HAND2 和 TWIST1 合作控制细胞周期进程。总的来说，我们发现 TCF4 通过直接转录调节驱动高风险 NB 的 c-MYC/MYCN 致癌程序来促进细胞增殖。从机制上讲，我们的数据表明，TCF4 通过将已知调节 MYC 功能的多种因子招募到关键 NB 转录因子、TCF4 和 MYC 癌蛋白之间的共定位位点来支持 MYC 活性。许多 TCF4 招募的因子都是可药物化的，有助于深入了解高风险 NB 的潜在疗法。这项研究确定了 I 类 bHLH 转录因子（例如 TCF3、TCF4 和 TCF12）的新功能，这些因子在癌症和发育中非常重要。© 2024 作者。约翰·威利出版的《分子肿瘤学》

Super-enhancer-associated transcription factor networks define cell identity in neuroblastoma (NB). Dysregulation of these transcription factors contributes to the initiation and maintenance of NB by enforcing early developmental identity states. We report that the class I basic helix-loop-helix (bHLH) transcription factor 4 (TCF4; also known as E2-2) is a critical NB dependency gene that significantly contributes to these identity states through heterodimerization with cell-identity-specific bHLH transcription factors. Knockdown of TCF4 significantly induces apoptosis in vitro and inhibits tumorigenicity in vivo. We used genome-wide expression profiling, TCF4 chromatin immunoprecipitation sequencing (ChIP-seq) and TCF4 immunoprecipitation-mass spectrometry to determine the role of TCF4 in NB cells. Our results, along with recent findings in NB for the transcription factors T-box transcription factor TBX2, heart- and neural crest derivatives-expressed protein 2 (HAND2) and twist-related protein 1 (TWIST1), propose a role for TCF4 in regulating forkhead box protein M1 (FOXM1)/transcription factor E2F-driven gene regulatory networks that control cell cycle progression in cooperation with N-myc proto-oncogene protein (MYCN), TBX2, and the TCF4 dimerization partners HAND2 and TWIST1. Collectively, we showed that TCF4 promotes cell proliferation through direct transcriptional regulation of the c-MYC/MYCN oncogenic program that drives high-risk NB. Mechanistically, our data suggest the novel finding that TCF4 acts to support MYC activity by recruiting multiple factors known to regulate MYC function to sites of colocalization between critical NB transcription factors, TCF4 and MYC oncoproteins. Many of the TCF4-recruited factors are druggable, giving insight into potential therapies for high-risk NB. This study identifies a new function for class I bHLH transcription factors (e.g., TCF3, TCF4, and TCF12) that are important in cancer and development.© 2024 The Author(s). Molecular Oncology published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.