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

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.

An inflammatory gene‐related prognostic risk score model for prognosis and immune infiltration in glioblastoma

“…39 EREG is the core onco-immunological biomarker of cuproptosis, and EREG knockdown can inhibit the proliferation, invasion, and migration of glioma cells. 40 To date, no relevant studies have been found on ADRM1 and IL18R1 in brain tumors. Mohan et al 41 reported that OSMR is upregulated in human brain tumor stem cells and correlated with poor prognosis.…”

“…A previous study showed that ABI1 is involved in the regulation of GBM cell invasion 39 . EREG is the core onco‐immunological biomarker of cuproptosis, and EREG knockdown can inhibit the proliferation, invasion, and migration of glioma cells 40 . To date, no relevant studies have been found on ADRM1 and IL18R1 in brain tumors.…”

Role of Epiregulin in Lung Tumorigenesis and Therapeutic Resistance

“…The oncogenic functions of EREG have been reported for various human cancers. Human recombinant EREG promotes cell proliferation in pancreatic [63] and bladder cancer [64] in a dose-dependent manner, whereas an EREG knockdown inhibits tumor growth in hepatocellular carcinoma [65], breast cancer [66], head and neck squamous cell carcinoma (HNSCC) [57], cervical cancer [54], glioma [67], and multiple myeloma [68]. Forced EREG expression enhances tumorigenicity in glioblastoma [58], salivary adenoid cystic carcinoma (SACC) [56], HNSCC [57], and esophageal cancer [59].…”

“…A recent study revealed that EREG is a core onco-immunological biomarker of cuproptosis [67], which is a molecular mechanism of regulated cell death [73]. Cuproptosis is induced by excessive copper, which binds to lipoylated dihydrolipoamide S-acetyltransferase (DLAT) [74].…”

“…Increasing evidence indicates the involvement of cuproptosis in the tumor progression and angiogenesis of lung cancer [75]. Recently, Zhou et al [67] reported that EREG influences glioma cell growth by affecting the process of cuproptosis, which is accompanied by the EREG-mediated upregulation of FDX1, a core regulatory protein in cuproptosis. These findings suggest that EREG is involved in cuproptosis by regulating FDX1, although the role of EREG in cuproptosis remains to be elucidated for lung cancer.…”