EREG 是细胞铜死亡的核心免疫生物标志物，介导了胶质母细胞瘤中 VEGF 和 CD99 信号的串扰。

EREG is the core onco-immunological biomarker of cuproptosis and mediates the cross-talk between VEGF and CD99 signaling in glioblastoma.

机构信息

Department of Neurosurgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Avenue, Wuhan, 430022, Hubei, China.

出版信息

J Transl Med. 2023 Jan 16;21(1):28. doi: 10.1186/s12967-023-03883-4.

DOI:10.1186/s12967-023-03883-4

PMID:36647156

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9843967/

Abstract

BACKGROUND

Glioma is the most prevalent primary tumor of the central nervous system. Glioblastoma multiforme (GBM) is the most malignant form of glioma with an extremely poor prognosis. A novel, regulated cell death form of copper-induced cell death called "cuproptosis" provides a new prospect for cancer treatment by regulating cuproptosis.

METHODS

Data from bulk RNA sequencing (RNA-seq) analysis (The Cancer Genome Atlas cohort and Chinese Glioma Genome Atlas cohort) and single cell RNA-seq (scRNA-seq) analysis were integrated to reveal their relationships. A scoring system was constructed according to the cuproptosis-related gene expression, and core genes were experimentally verified using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), Western blot (WB), immunohistochemistry (IHC), and immunofluorescence (IF). Moreover, cell counting kit-8 (CCK8), colony formation, 5-ethynyl-2'-deoxyuridine (EdU) incorporation, transwell, and flow cytometry cell cycle were performed to evaluate cell proliferation, invasion, and migration.

RESULTS

The Cuproptosis Activation Scoring (CuAS) Model has stable and independent prognostic efficacy, as verified by two CGGA datasets. Epiregulin (EREG), the gene of the model has the most contributions in the principal component analysis (PCA), is an onco-immunological gene that can affect immunity by influencing the expression of programmed death-ligand 1 (PD-L1) and mediate the process of cuproptosis by influencing the expression of ferredoxin 1 (FDX1). Single cell transcriptome analysis revealed that high CuAS GBM cells are found in vascular endothelial growth factor A (VEGFA) + malignant cells. Oligodendrocyte transcription factor 1 (OLIG1) + malignant is the original clone, and VEGF and CD99 are the differential pathways of specific cell communication between the high and low CuAS groups. This was also demonstrated by immunofluorescence in the tissue sections. Furthermore, CuAS has therapeutic potential for immunotherapy, and we predict that many drugs (methotrexate, NU7441, KU -0063794, GDC-0941, cabozantinib, and NVP-BEZ235) may be used in patients with high CuAS.

CONCLUSION

EREG is the core onco-immunological biomarker of CuAS and modulates the cross-talk between VEGF and CD99 signaling in glioblastoma, and CuAS may provide support for immunotherapy and chemotherapy.

摘要

背景

神经胶质瘤是中枢神经系统最常见的原发性肿瘤。多形性胶质母细胞瘤（GBM）是最恶性的神经胶质瘤形式，预后极差。一种新的、受调控的铜诱导细胞死亡形式称为“铜死亡”，通过调控铜死亡为癌症治疗提供了新的前景。

方法

整合来自批量 RNA 测序（RNA-seq）分析（癌症基因组图谱队列和中国神经胶质瘤基因组图谱队列）和单细胞 RNA-seq（scRNA-seq）分析的数据，以揭示它们之间的关系。根据铜死亡相关基因表达构建评分系统，并使用实时定量逆转录聚合酶链反应（qRT-PCR）、Western blot（WB）、免疫组织化学（IHC）和免疫荧光（IF）实验验证核心基因。此外，通过细胞计数试剂盒-8（CCK8）、集落形成、5-乙炔基-2'-脱氧尿苷（EdU）掺入、transwell 和流式细胞术细胞周期评估细胞增殖、侵袭和迁移。

结果

Cuproptosis Activation Scoring（CuAS）模型具有稳定且独立的预后效能，这在两个 CGGA 数据集得到了验证。模型的基因表皮调节素（EREG）在主成分分析（PCA）中贡献最大，是一种癌免疫基因，可通过影响程序性死亡配体 1（PD-L1）的表达来影响免疫，并通过影响铁氧还蛋白 1（FDX1）的表达来介导铜死亡过程。单细胞转录组分析表明，高 CuAS GBM 细胞存在于血管内皮生长因子 A（VEGFA）+恶性细胞中。少突胶质细胞转录因子 1（OLIG1）+恶性是原始克隆，VEGF 和 CD99 是高低 CuAS 组之间特定细胞通讯的差异途径。这也通过组织切片的免疫荧光得到了证明。此外，CuAS 对免疫治疗具有治疗潜力，我们预测许多药物（甲氨蝶呤、NU7441、KU-0063794、GDC-0941、卡博替尼和 NVP-BEZ235）可能用于高 CuAS 患者。