垂体神经内分泌瘤(PitNETs)起源于垂体腺，占所有颅内肿瘤的10%-15%。近期研究表明，增强子RNA(eRNA)在肿瘤生长中起调控作用。然而，eRNA调控PitNETs致瘤机制尚未阐明。通过使用正常垂体和PitNETs组织，鉴定差异表达的eRNAs(DEEs)。根据单个样本基因集富集分析(ssGSEA)算法，利用GSVA测定免疫基因集和癌症相关基因集。通过CIBERSORT算法计算所有样本中免疫细胞的分布。此外，通过Pearson相关分析构建关键DEEs、eRNAs靶基因、癌症相关基因集、差异表达转录因子、免疫细胞和免疫基因集的调控网络。通过CMap分析探索小分子抗PitNETs药物并通过体外和体内实验、ATAC-seq和ChIP-seq验证研究的准确性。 本研究从公共数据库中检索了134个PitNETs和107个非肿瘤垂体样本的数据，以鉴定差异表达基因。总共鉴定出1128个差异表达eRNAs(DEEs)（494个上调eRNAs和634个下调eRNAs）。随后，检验DEEs与与癌症相关和免疫相关基因标志物的相关性，建立了一个共表达调控网络，其中包括18个DEEs、50个潜在的DEEs目标基因、5个癌症标志基因集、2个差异表达转录因子、4种免疫细胞类型和4个免疫基因集。基于该网络，通过Connectivity Map分析鉴定了以下四个用于PitNETs的治疗药物：环可吡索、倍普利、氯米帕明和甲硝啶。这些药物的抑制生长效果通过体外实验得到验证。环可吡索对PitNETs具有潜在的抑制生长效果。在DEEs中，GNLY、HOXB7、MRPL33、PRDM16、TCF7和ZNF26被确定为PitNETs的潜在诊断和治疗生物标志物。 本研究揭示了eRNAs在PitNETs发生和发展中的重要影响。通过构建共表达调控网络，我们鉴定了GNLY、HOXB6、MRPL33、PRDM16、TCF7和ZNF26等相对重要的DEEs，这些DEEs被认为是PitNETs诊断和治疗的新生物标志物。本研究证实了eRNAs在PitNETs发生和发展中的作用，并发现环可吡索是垂体腺瘤的潜在治疗药物。 版权所有 © 2023 Wang, Wei, Wang, Huang, Zhang, Dai, Xue, Lin和Wu。

Pituitary neuroendocrine tumors (PitNETs), which originate from the pituitary gland, account for 10%-15% of all intracranial neoplasms. Recent studies have indicated that enhancer RNAs (eRNAs) exert regulatory effects on tumor growth. However, the mechanisms underlying the eRNA-mediated tumorigenesis of PitNETs have not been elucidated.Normal pituitary and PitNETs tissues were used to identify the differentially expressed eRNAs (DEEs). Immune gene sets and hallmarks of cancer gene sets were quantified based on single sample gene set enrichment analysis (ssGSEA) algorithm using GSVA. The perspective of immune cells among all samples was calculated by the CIBERSORT algorithm. Moreover, the regulatory network composed of key DEEs, target genes of eRNAs, hallmarks of cancer gene sets, differentially expressed TF, immune cells and immune gene sets were constructed by Pearson correlation analysis. Small molecular anti-PitNETs drugs were explored by CMap analysis and the accuracy of the study was verified by in vitro and in vivo experiments, ATAC-seq and ChIP-seq.In this study, data of 134 PitNETs and 107 non-tumorous pituitary samples were retrieved from a public database to identify differentially expressed genes. In total, 1128 differentially expressed eRNAs (DEEs) (494 upregulated eRNAs and 634 downregulated eRNAs) were identified. Next, the correlation of DEEs with cancer-related and immune-related gene signatures was examined to establish a co-expression regulatory network comprising 18 DEEs, 50 potential target genes of DEEs, 5 cancer hallmark gene sets, 2 differentially expressed transcription factors, 4 immune cell types, and 4 immune gene sets. Based on this network, the following four therapeutics for PitNETs were identified using Connectivity Map analysis: ciclopirox, bepridil, clomipramine, and alexidine. The growth-inhibitory effects of these therapeutics were validated using in vitro experiments. Ciclopirox exerted potential growth-inhibitory effects on PitNETs. Among the DEEs, GNLY, HOXB7, MRPL33, PRDM16, TCF7, and ZNF26 were determined to be potential diagnostic and therapeutic biomarkers for PitNETs.This study illustrated the significant influence of eRNAs on the occurrence and development of PitNETs. By constructing the co-expression regulation network, GNLY, HOXB6, MRPL33, PRDM16, TCF7, and ZNF26 were identified as relatively significant DEEs which were considered as the novel biomarkers of diagnosis and treatment of PitNETs. This study demonstrated the roles of eRNAs in the occurrence and development of PitNETs and revealed that ciclopirox was a potential therapeutic for pituitary adenomas.Copyright © 2023 Wang, Wei, Wang, Huang, Zhang, Dai, Xue, Lin and Wu.