Investigating the Association between the Autophagy Markers LC3B, SQSTM1/p62, and DRAM and Autophagy-Related Genes in Glioma

Background: Glioma remains challenging due to high recurrence rates and resistance to treatment. Diagnosis and follow-up in resource-constrained regions often leads to significant patient attrition. Serum microRNA (miRNA) expression profiles, which have been shown to correlate with tissue expression profiles, are detectable in peripheral blood samples, providing a promising avenue for non-invasive and repeatable liquid biopsies. miR-21 shows promise in many populations; however, there is a dearth of data from our region. Methodology: We collected 90 tumor tissues, 42 pre- and post-operative serum samples from glioma patients, and included 10 normal tissue adjacent to the tumor (NATs) along with serum samples from 8 healthy individuals and analyzed for miR-21 expression through RT-qPCR. Shapiro Wilk test was applied to calculate data distribution, ANOVA, Fisher's exact and Wilcox test, along with pairwise Student's t-test, were applied to determine the differences in gene expression. The expression level of miR-21 was assessed for correlation with Kaplan-Meier survival curves and different molecular markers (IDH, Ki-67, ATRX and p53). The quantitative hazard ratio was determined using Cox regression analysis. Results: miR-21 expression in tissue increased with the median fold expression of 2.35 in grade 2, 7.49 in grade 3 and 26 in grade 4 for glioma patients. The expression level showed significant difference between control tissue and grade 4 patients along with significant inter-comparison between grade 1 and grade 4, as well as grade 2 and grade 4. A 30-fold elevated expression of miR-21 has been noticed in patients above 50 years of age. Similarly, in serum samples a significant decline in miR-21 expression was observed in post-operative samples as compared to pre-operative samples in grade 2 (11-fold), grade 3 15-fold) and grade 4 (13-fold). Furthermore, there was positive correlation of miR-21 expression with tumor volume. IDH-wildtype and high Ki-67 expression in gliomas showed significant upregulation of miR-21 compared to IDH-mutant and low Ki-67 respectively. Patients with low miR-21 expression had significantly longer overall survival (OS) than patients with high miR-21 expression. Quantitative hazard analysis indicates that patients in the high expression group have a 3.4 times higher risk of mortality (95% CI: 1.6- 7.1), in comparison to patients in the low expression group with AUC of 0.742 (all p <0.05). Conclusion: This study demonstrates the potential of microRNA 21 as a serum biomarker for early, cost-effective diagnosis of glioma. Furthermore, it may inform the development of targeted treatment strategies for various glioma grades, particularly in our population.

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Autophagy-associated biomarkers ULK2, UVRAG, and miRNAs miR-21, miR-126, and miR-374: Prognostic significance in glioma patients

Amin,

Enam,

Sufiyan

et al. 2024

PLoS ONE

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As the pioneering study from Pakistan, our research distinctly focuses on validating the roles of autophagy-associated genes and MicroRNAs (miRs) in the unique context of our population for glioma prognosis. The study delves into the nuanced interplay of autophagy within a miR-modulated environment, prompting an exploration of its potential impact on glioma development and survival. Employing real-time PCR (qPCR), we meticulously assessed the expression profiles of autophagy genes and miRs in glioma tissues, complemented by immunohistochemistry on Formalin-fixed paraffin-embedded tissues from the same patients. Our comprehensive statistical analyses, including the data normality hypothesis Shapiro-Wilk test, the Mann-Whitney U-test, Spearman correlation test, and Kaplan-Meier survival analysis, were tailored to unravel the intricate associations specific to low- and high-grade glioma within our population. Clinicopathological analysis revealed a predominance of male patients (66%) with a median age of 35 years. Glioblastoma (32%) and Astrocytoma (36%) were the most prevalent histopathological subtypes. Molecular analysis showed significant correlations between prognostic markers (Ki-67, IDH-1, p53) and clinicopathological factors, including age, histological type, radiotherapy, and chemotherapy. In high-grade glioma, increased expression of AKT and miR-21, coupled with reduced ULK2 and LC3 expression was distinctly observed. While correlation analysis identified a strong positive correlation between ULK2 and UVRAG, PTEN, miR-7, and miR-100 in low-grade glioma, unveiling distinctive molecular signatures unique to our study. Furthermore, a moderate positive correlation emerged between ULK2 and mTOR, miR-7, miR-30, miR-100, miR-204, and miR-374, also between miR-21 and miR-126. Similarly, a positive correlation appeared between ULK2 and AKT, LC3, PI3K, PTEN, ULK1, VPS34, mTOR, Beclin1, UVRAG, miR-7 and miR-374. AKT positively correlated with LC3, PI3K, PTEN, ULK1, VPS34, mTOR, Beclin1, UVRAG, miR-7, miR-30, miR-204, miR-374, miR-126 and miR-21 weakly correlated with AKT and miR-30 in high-grade glioma, providing further insights into the autophagy pathway within our population. The enrichment analysis for miR-21, miR-126, and miR-374 showed MAPK pathway as a common pathway along with Ras, PI3K, and mTOR pathway. The low ULK2, UVRAG, and miR-374 expression group exhibited significantly poor overall survival in glioma, while miR-21 over-expression indicated a poor prognosis in glioma patients, validating it in our population. This study provides comprehensive insights into the molecular landscape of gliomas, highlighting the dysregulation of autophagy genes ULK2, and UVRAG and the associated miR-21, miR-126 and miR-374 as potential prognostic biomarkers and emphasizing their unique significance in shaping survival outcomes in gliomas within the specific context of the Pakistani population.

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Investigating the Association between the Autophagy Markers LC3B, SQSTM1/p62, and DRAM and Autophagy-Related Genes in Glioma

Cited by 3 publications

References 53 publications

LC3B: A microtubule-associated protein influences disease progression and prognosis

LC3B: A microtubule-associated protein influences disease progression and prognosis

Clinical and Molecular Characterization of Glioma Patients: miR-21 Expression as a Prognostic Biomarker in Tissue and Serum

Autophagy-associated biomarkers ULK2, UVRAG, and miRNAs miR-21, miR-126, and miR-374: Prognostic significance in glioma patients

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