Key Genes Involved in the Beneficial Mechanism of Hyperbaric Oxygen for Glioblastoma and Predictive Indicators of Hyperbaric Oxygen Prolonging Survival in Glioblastoma Patients

Abstract

Objective

The prognosis of glioblastoma is poor, and therapy-resistance is largely attributed to intratumor hypoxia. Hyperbaric oxygen (HBO) effectively alleviates hypoxia. However, the sole role of HBO in glioblastoma remains controversial. We previously reported that HBO can promote apoptosis, shorten protrusions, and delay growth of glioblastoma, but the molecular mechanism is unclear. We aimed to test candidate genes in HBO-exposed glioblastoma cells and to analyze their correlation with the survival of glioblastoma patients.

Methods

Glioblastoma cell lines exposed to repetitive HBO or normobaric air (NBA) were collected for RNA isolation and microarray data analysis. GO analysis, KEGG pathway analysis and survival analysis of the differentially expressed genes (DEGs) were performed.

Results

HBO not only inhibited hypoxia-inducing genes including CA9, FGF11, PPFIA4, TCAF2 and SLC2A12, but also regulated vascularization by downregulating the expression of COL1A1, COL8A1, COL12A1, RHOJ and FILIP1L, ultimately attenuated hypoxic microenvironment of glioblastoma. HBO attenuated inflammatory microenvironment by reducing the expression of NLRP2, CARD8, MYD88 and CD180. HBO prevented metastasis by downregulating the expression of NTM, CXCL12, CXCL13, CXCR4, CXCR5, CDC42, IGFBP3, IGFBP5, GPC6, MMP19, ADAMTS1, EFEMP1, PTBP3, NF1 and PDCD1. HBO upregulated the expression of BAK1, PPIF, DDIT3, TP53I11 and FAS, whereas downregulated the expression of MDM4 and SIVA1, thus promoting apoptosis. HBO upregulated the expression of CDC25A, MCM2, PCNA, RFC33, DSCC1 and CDC14A, whereas downregulated the expression of ASNS, CDK6, CDKN1B, PTBP3 and MAD2L1, thus inhibiting cell cycle progression. Among these DEGs, 17 indicator-genes of HBO prolonging survival were detected.

Conclusions

HBO is beneficial for glioblastoma. Glioblastoma patients with these predictive indicators may prolong survival with HBO therapy. These potential therapeutic targets especially COL1A1, ADAMTS1 and PTBP3 deserve further validation.

References

[1] Louis DN, Perry A, Wesseling P, et al.. The 2021 WHO Classification of Tumors of the Central Nervous System: a summary. Neuro Oncol, 2021, 23(8): 1231-1251

[2] Dubinski D, Hattingen E, Senft C, et al.. Controversial roles for dexamethasone in glioblastoma - Opportunities for novel vascular targeting therapies. J Cereb Blood Flow Metab, 2019, 39(8): 1460-1468

[3] Stupp R, Mason WP, van den Bent MJ, et al.. European Organisation for Research and Treatment of Cancer Brain Tumor and Radiotherapy Groups; National Cancer Institute of Canada Clinical Trials Group. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med, 2005, 352(10): 987-996

[4] Zhou Y, Zhou Y, Shingu T, et al.. Metabolic alterations in highly tumorigenic glioblastoma cells: preference for hypoxia and high dependency on glycolysis. J Biol Chem, 2011, 286(37): 32843-32853

[5] Bar EE. Glioblastoma, cancer stem cells and hypoxia. Brain Pathol, 2011, 21(2): 119-129

[6] Liang BC. Effects of hypoxia on drug resistance phenotype and genotype in human glioma cell lines. J Neurooncol, 1996, 29(2): 149-155

[7] Jensen RL. Hypoxia in the tumorigenesis of gliomas and as a potential target for therapeutic measures. Neurosurg Focus, 2006, 20(4): E24

[8] Huang L, Boling W, Zhang JH. Hyperbaric oxygen therapy as adjunctive strategy in treatment of glioblastoma multiforme. Med Gas Res, 2018, 8(1): 24-28

[9] Pan JQ, Tian ZM, Xue LB. Hyperbaric Oxygen Treatment for Long COVID: From Molecular Mechanism to Clinical Practice. Curr Med Sci, 2023, 43(6): 1061-1065

[10] Beppu T, Kamada K, Yoshida Y, et al.. Change of oxygen pressure in glioblastoma tissue under various conditions. J Neurooncol, 2002, 58(1): 47-52

[11] Lu Z, Ma J, Liu B, et al.. Hyperbaric oxygen therapy sensitizes nimustine treatment for glioma in mice. Cancer Med, 2016, 5(11): 3147-3155

[12] Sun S, Lee D, Lee NP, et al.. Hyperoxia resensitizes chemoresistant human glioblastoma cells to temozolomide. J Neurooncol, 2012, 109(3): 467-475

[13] Dagıstan Y, Karaca I, Bozkurt ER, et al.. Combination hyperbaric oxygen and temozolomide therapy in C6 rat glioma model. Acta Cir Bras, 2012, 27(6): 383-387

[14] Zeng X, Wang Q, Tan X, et al.. Mild thermotherapy and hyperbaric oxygen enhance sensitivity of TMZ/PSi nanoparticles via decreasing the stemness in glioma. J Nanobiotechnology, 2019, 17(1): 47

[15] Stuhr LE, Raa A, Oyan AM, et al.. Hyperoxia retards growth and induces apoptosis, changes in vascular density and gene expression in transplanted gliomas in nude rats. J Neurooncol, 2007, 85(2): 191-202

[16] Yuen CM, Tsai HP, Tseng TT, et al.. Hyperbaric Oxygen Therapy Adjuvant Chemotherapy and Radiotherapy through Inhibiting Stemness in Glioblastoma. Curr Issues Mol Biol, 2023, 45(10): 8309-8320

[17] Wang YG, Zhan YP, Pan SY, et al.. Hyperbaric oxygen promotes malignant glioma cell growth and inhibits cell apoptosis. Oncol Lett, 2015, 10(1): 189-195

[18] Wang YG, Long J, Shao DC, et al.. Hyperbaric oxygen inhibits production of CD3+ T cells in the thymus and facilitates malignant glioma cell growth. J Int Med Res, 2018, 46(7): 2780-2791

[19] Wang P, Gong S, Pan J, et al.. Hyperbaric oxygen promotes not only glioblastoma proliferation but also chemosensitization by inhibiting HIF1α/HIF2α-Sox2. Cell Death Discov, 2021, 7(1): 103

[20] D’Agostino DP, Olson JE, Dean JB. Acute hyperoxia increases lipid peroxidation and induces plasma membrane blebbing in human U87 glioblastoma cells. Neuroscience, 2009, 159(3): 1011-1022

[21] Yu QH, Li DG, Zhang Q, et al.. Effect of hyperbaric oxygen on survival of human glioma 172 cells. Chin J Rehab, 2020, 35(6): 283-286

[22] Wang DW, Su F, Yang LJ, et al.. Bioinformatics Analysis and Identification of Potential Genes Associated with Pathogenesis and Prognosis of Gastric Cancer. Curr Med Sci, 2022, 42(2): 357-372

[23] Zhao Z, Zhang KN, Wang Q, et al.. Chinese Glioma Genome Atlas (CGGA): A Comprehensive Resource with Functional Genomic Data from Chinese Glioma Patients. Genomics Proteomics Bioinformatics, 2021, 19(1): 1-12

[24] Wu Q, Zhang B, Li B, et al.. PTBP3 promotes migration of non-small cell lung cancer through regulating E-cadherin in EMT signaling pathway. Cancer Cell Int, 2020, 20: 172

[25] Arienti C, Pignatta S, Zanoni M, et al.. High-pressure oxygen rewires glucose metabolism of patient-derived glioblastoma cells and fuels inflammasome response. Cancer Lett, 2021, 506: 152-166

[26] Chen Y, Ji Y, Liu S, et al.. PTBP3 regulates proliferation of lung squamous cell carcinoma cells via CDC25A-mediated cell cycle progression. Cancer Cell Int, 2022, 22(1): 19

[27] Beppu T, Kamada K, Nakamura R, et al.. A phase II study of radiotherapy after hyperbaric oxygenation combined with interferon-β and nimustine hydrochloride to treat supratentorial malignant gliomas. J Neurooncol, 2003, 61: 161-170

[28] Comba A, Faisal SM, Dunn PJ, et al.. Spatiotemporal analysis of glioma heterogeneity reveals COL1A1 as an actionable target to disrupt tumor progression. Nat Commun, 2022, 13(1): 3606

[29] Ma J, Weng L, Jia Y, et al.. PTBP3 promotes malignancy and hypoxia-induced chemoresistance in pancreatic cancer cells by ATG12 up-regulation. J Cell Mol Med, 2020, 24(5): 2917-2930

[30] Xie P, Zhang Y, Chen R, et al.. PTBP3 promotes tumorigenesis of glioblastoma by stabilizing Twist1. Transl Oncol, 2022, 25: 101520