In silico, in vitro, and ex vivo analysis reveals miR-27a-3p and miR-155-5p as key microRNAs for glioblastoma progression: Insights into Th1 differentiation and apoptosis induction

• Published in: The FASEB journal 2024-12, Vol.38 (24), p.e70255
• Main Authors: Weber, Augusto Ferreira, Scholl, Juliete Nathali, Dias, Camila Kehl, Lima, Vinícius Pierdoná, Assmann, Taís Silveira, Anzolin, Eduardo, Kus, Willian Pegoraro, Worm, Paulo Valdeci, Battastini, Ana Maria Oliveira, Figueiró, Fabrício
• Format: Article
• Language: English
• Subjects: Apoptosis - genetics; Brain Neoplasms - genetics; Brain Neoplasms - metabolism; Brain Neoplasms - pathology; Cell Differentiation; Cell Line, Tumor; Computer Simulation; Disease Progression; Gene Expression Regulation, Neoplastic; Glioblastoma - genetics; Glioblastoma - metabolism; Glioblastoma - pathology; Humans; MicroRNAs - genetics; MicroRNAs - metabolism; Th1 Cells - immunology; Th1 Cells - metabolism
• ISSN: 1530-6860; 1530-6860
• DOI: 10.1096/fj.202401538R

We explored key microRNAs (miRNAs) related to tumorigenesis and immune modulation in glioblastoma (GBM), employing in silico, in vitro, and ex vivo analysis along with an assessment of the cellular impacts resulting from miRNA inhibition. GBM and T cells miRNA expression profiles from public datasets were used to evaluate differentially expressed miRNAs (DEmiRNAs). Some DEmiRNAs were chosen for validation in GBM cell lines, primary cell cultures, and brain tumor patient samples, using RT-qPCR. Target genes and pathways were identified with bioinformatic analyses. In silico functional enrichment analysis revealed that miR-27a-3p and miR-155-5p modulate immune, metabolic, and GBM-related pathways. A172 cells were transfected with miRNA inhibitors and the effects on cellular processes and immunomodulation were analyzed by co-culture assays and flow cytometry. Upon validation, miR-27a-3p and miR-155-5p miRNAs expressions were consistently increased. Inhibiting these two miRNAs reduced cell viability, but only the inhibition of miR-27a-3p led to apoptosis. Co-culture assays showed an increase in Th1 cells along with elevated Th1/Treg and Th17/Treg ratios, and an increase in Th17 cells exclusively with miR-155-5p inhibition. Immune cells' gene expression modulation induced an antitumor profile, concomitant with an increase in the expression of apoptotic genes in cancer cells after co-culture. This study unveils potential targets for immune and tumor regulation, highlighting overexpressed miRNAs modulation as a novel therapeutic approach for GBM.