Journal: Scientific Reports

Article Title: New microRNA-based therapies reveal common targets in paediatric medulloblastoma and adult glioblastoma

doi: 10.1038/s41598-025-05517-9

Figure Lengend Snippet: Predictive Sfold analysis illustrating the binding sites between miR-206 and CORO1C and miR-383-5p and SV2B. ( a ) 3′UTR of CORO1C and miR-206 binding at site position 1614–1639, ΔG hybrid = − 23.4 kcal/mol. ( b ) 3′UTR of CORO1C and miR-206 binding at site position 1655–1675, ΔG hybrid = − 20.6 kcal/mol. ( c ) 3′UTR of SV2B and miR-383-5p binding at site position 3239–3263, ΔG hybrid = − 20.3 kcal/mol.

Article Snippet: Following this, the PVDF membranes were incubated with TBST and the appropriate primary antibody antisera against CORO1C (dilution 1:500), SV2B (dilution 1:500), and β-actin (dilution 1:1000) overnight at 4 °C (ProteinTech, Manchester, UK).

Techniques: Binding Assay

Journal: Scientific Reports

Article Title: New microRNA-based therapies reveal common targets in paediatric medulloblastoma and adult glioblastoma

doi: 10.1038/s41598-025-05517-9

Figure Lengend Snippet: Comprehensive transcriptomics analysis of MB tissue profiles. ( a ) Heatmap of dysregulated genes (control group D (D); MB group C (C1-C9). ( b ) Volcano plot of gene expression revealed 1013 dysregulated genes: 415 downregulated and 598 upregulated. CORO1C and SV2B were identified as upregulated in the patient sample cohort. ( c ) Pathway enrichment analysis revealed that DEG were primarily associated with the regulation of the cell cycle, synaptic vesicle cycle, axon guidance, and miRNAs in cancer (red boxes). ( d ) The molecular function analysis showed that the vast majority of DEG were involved in processes such as ATP binding and nucleic acid binding (red boxes). ( e ) Biological process analysis showed that most dysregulated genes were involved cell division, cellular response to DNA damage, and mitotic cell cycle checkpoints (red boxes). Transcriptomic and sRNA-seq data were generated from the same patient cohort (n = 9). For transcriptomics analyses, p-values were adjusted using the Benjamini–Hochberg method to control the false discovery rate. Genes with an adjusted p-value < 0.05 were considered differentially expressed. A q-value < 0.005 and |log₂(fold change)|≥ 1 were used as thresholds to define significant differential expression, capturing genes with meaningful and potentially biologically relevant changes, even below the more stringent fold change cutoff of > 2.

Article Snippet: Following this, the PVDF membranes were incubated with TBST and the appropriate primary antibody antisera against CORO1C (dilution 1:500), SV2B (dilution 1:500), and β-actin (dilution 1:1000) overnight at 4 °C (ProteinTech, Manchester, UK).

Techniques: Control, Gene Expression, Binding Assay, Generated, Quantitative Proteomics

Journal: Scientific Reports

Article Title: New microRNA-based therapies reveal common targets in paediatric medulloblastoma and adult glioblastoma

doi: 10.1038/s41598-025-05517-9

Figure Lengend Snippet: RT-qPCR validations within MB and GB cell lines. ( a ) miR-206 was significantly downregulated in MB cells in comparison to normal human astrocytes (p < 0.0001). ( b ) miR-383 was significantly downregulated in MB cells in comparison to normal human astrocytes (p < 0.0096). ( c ) CORO1C was significantly upregulated in MB cells in comparison to normal human astrocytes (p < 0.0218). ( d ) SV2B was significantly upregulated in MB cells in comparison to normal human astrocytes (p < 0.0001). ( e ) miR-206 was significantly downregulated in U87MG and U251MG GB cells in comparison to normal human astrocytes (U87MG, p < 0.0033; U251MG, p < 0.0024). ( f ) miR-383 was significantly downregulated in GB cells in comparison to normal human astrocytes (U87MG, p < 0.0002; U251MG p < 0.0009). ( g ) CORO1C was significantly upregulated in GB cells in comparison to normal human astrocytes (U251MG, p < 0.0002; U87MG p < 0.0001). ( h ) SV2B was significantly upregulated in GB cells in comparison to normal human astrocytes (U251MG, p < 0.0001; U87MG p < 0.0001). Results are based on three independent experiments, n = 3. Statistical analyses were conducted using ordinary one-way ANOVA, followed by Dunnett’s multiple comparisons test. When only two groups were compared, unpaired t-tests were used to determine significant differences in expression. Error bars represent the standard deviation (SD) of the mean. ns non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Following this, the PVDF membranes were incubated with TBST and the appropriate primary antibody antisera against CORO1C (dilution 1:500), SV2B (dilution 1:500), and β-actin (dilution 1:1000) overnight at 4 °C (ProteinTech, Manchester, UK).

Techniques: Quantitative RT-PCR, Comparison, Expressing, Standard Deviation

Journal: Scientific Reports

Article Title: New microRNA-based therapies reveal common targets in paediatric medulloblastoma and adult glioblastoma

doi: 10.1038/s41598-025-05517-9

Figure Lengend Snippet: Immunofluorescence and western blot analysis of CORO1C and SV2B within HA-bs and MB cells, alongside CORO1C IF analysis within GB cells. ( a ) IF staining revealed that CORO1C, SV2B, and β-actin exhibited uniform staining across normal human astrocytes, with CORO1C showing nuclear localisation and SV2B showing cytoplasmic localisation. ( b ) IF staining within DAOY MB cells showed an increased fluorescence of SV2B in MB cells when compared to HA-bs cells, alongside a nucellar localisation of CORO1C similar to the one observed in HA-bs cells. ( c ) IF staining within GB cells revealed that there was increased fluorescence of CORO1C within both, U251MG and U87MG cells in comparison to HA-bs cells. ( d ) Chemiluminescent images showing the expression of β-actin, CORO1C, and SV2B within HA-bs, U251MG, U87MG, and DAOY cells. ( e ) CORO1C protein expression was significantly upregulated within all GB and MB cell lines in comparison to normal HA-bs cells (U251MG, p < 0.0001; U87MG, p < 0.003; DAOY, p < 0.0002). ( f ) SV2B protein expression was significantly upregulated within U251MG, U87MG, and DAOY cells in comparison to HA-bs cells (p < 0.037, p < 0.003, p < 0.0001, respectively). ( g ) β-actin protein expression revealed no significant difference between the normal astrocytes and GB and MB cells. Results are based on three independent experiments, n = 3. All statistical analyses were conducted using ordinary one-way ANOVA, followed by Dunnett’s multiple comparisons test to determine the significant differences in expression between normal human astrocytes, GB and MB cells. Error bars represent the SD of the mean. Original blots/gels are presented in Supplementary Fig. . ns non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Following this, the PVDF membranes were incubated with TBST and the appropriate primary antibody antisera against CORO1C (dilution 1:500), SV2B (dilution 1:500), and β-actin (dilution 1:1000) overnight at 4 °C (ProteinTech, Manchester, UK).

Techniques: Immunofluorescence, Western Blot, Staining, Fluorescence, Comparison, Expressing

Journal: Scientific Reports

Article Title: New microRNA-based therapies reveal common targets in paediatric medulloblastoma and adult glioblastoma

doi: 10.1038/s41598-025-05517-9

Figure Lengend Snippet: Immunohistochemistry analyses of CORO1C. ( a ) All age groups showed significantly increased levels of CORO1C in comparison to their normal counterparts. The age groups of 0–20 years showed the most significantly increased levels of CORO1C in comparison to normal and normal adjacent tissues controls (Normal Tissue versus Age 0–20, p < 0.0001; Adjacent brain tissue versus Age 0–20, p < 0.0001; Normal Tissue versus Age 21–40, p < 0.0009; Adjacent Brain Tissue versus Age 21–40, p < 0.0001; Normal Tissue versus Age 41–60, p < 0.0019; Adjacent Brain Tissue versus Age 41–60, p < 0.0001; Normal Tissue versus Age 61–80, p < 0.0360; Adjacent Brain Tissue versus Age 61–80, p < 0.0012). ( b ) Significantly higher levels of the CORO1C protein were observed within MB and GB, in comparison to the rest of the malignancies and their normal counterparts (Adjacent Brain Tissue versus Astrocytoma, p < 0.0014; Normal Tissue versus Ependymoma, p < 0.0027; Adjacent Brain Tissue versus Ependymoma, p < 0.0001; Adjacent Brain Tissue versus Oligodendroglioma, p < 0.0037; Normal Tissue versus MB, p < 0.0001; Adjacent Brain Tissue versus MB, p < 0.0001; Normal Tissue versus GB, p < 0.0001; Adjacent Tissue versus GB, p < 0.0001). ( c ) Significant differences between tumour grades 2, 3 and 4 in comparison to normal brain tissue were observed. Grade 1 tumours showed significantly higher expression in comparison to normal adjacent tissue. Significant differences between grades 3 and 4 and grades 1 and 2 (Adjacent Brain Tissue versus Grade 1, p < 0.0460; Normal Tissue versus Grade 2, p < 0.0395; Adjacent Brain Tissue versus Grade 2, p < 0.0009; Normal Tissue versus Grade 3, p < 0.0003; Adjacent Brain Tissue versus Grade 3, p < 0.0001; Normal Tissue versus Grade 4, p < 0.0001; Adjacent Brain Tissue versus Grade 4, p < 0.0001; Grade 1 versus Grade 3, p < 0.0012; Grade 1 versus Grade 4, p < 0.0001; Grade 2 versus Grade 4, p < 0.0001). ( d ) Microscopic images of CORO1C stained tissues sections illustrating the staining intensity within normal brain tissue, tumour adjacent tissue, MB and GB tissues at × 100 magnification. ( e ) Microscopic images of CORO1C stained tissues cores of normal brain tissue, tumour adjacent tissue, and different brain malignancies at × 40 magnification. Results are based on three independent scoring assessments, n = 3. All statistical analyses were conducted using ordinary one-way ANOVA, followed by Dunnett’s multiple comparisons test to determine the significant differences in CORO1C expression. Error bars represent the SD of the mean. ns non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, NT normal tissue, NAT normal adjacent tissue, AST astrocytoma, MB medulloblastoma, GB glioblastoma.

Article Snippet: Following this, the PVDF membranes were incubated with TBST and the appropriate primary antibody antisera against CORO1C (dilution 1:500), SV2B (dilution 1:500), and β-actin (dilution 1:1000) overnight at 4 °C (ProteinTech, Manchester, UK).

Techniques: Immunohistochemistry, Comparison, Expressing, Staining

Journal: Scientific Reports

Article Title: New microRNA-based therapies reveal common targets in paediatric medulloblastoma and adult glioblastoma

doi: 10.1038/s41598-025-05517-9

Figure Lengend Snippet: Transient transfections of MB cells. ( a, b ) miR-206 and miR-383 were significantly upregulated after transfections with miR-206 and miR-383 mimics (miR-206, p < 0.0001; miR-383, p < 0.0001). ( c ) CORO1C was significantly downregulated post transfection with miR-206 mimic (p < 0.0009). ( d ) SV2B was significantly downregulated post transfection with miR-383 mimic (p < 0004). ( e, f ) MB cell viability was significantly decreased post transfection with miR-206 and miR-383 mimics (miR-206, p < 0.0032; miR-383, p < 0.009). ( g, h ) MB cells lost their colony forming abilities post transfections with miR-206 and miR-383 mimics. ( i ) Microscopic images of CF within MB cells post exposure to miR-206 and miR-383 mimics (× 40 magnification). ( j ) Western blotting analysis showed that the protein levels of CORO1C were significantly decreased in comparison to the negative control condition post exposure to miR-206 mimic (p < 0.0001). The protein levels of SV2B were also significantly reduced in comparison to the negative control condition post exposure to miR-383 mimic (p < 0.0009). ( k ) Chemiluminescent images showing the expression of β-actin, CORO1C, and SV2B within negative control miRNA #1, positive control miR-1, and miR-206 and miR-383 mimics. Results are based on three independent experiments, n = 3. All statistical analyses were conducted using ordinary one-way ANOVA, followed by Dunnett’s multiple comparisons test. Error bars represent the SD of the mean. Original blots/gels are presented in Supplementary Fig. . ns non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Following this, the PVDF membranes were incubated with TBST and the appropriate primary antibody antisera against CORO1C (dilution 1:500), SV2B (dilution 1:500), and β-actin (dilution 1:1000) overnight at 4 °C (ProteinTech, Manchester, UK).

Techniques: Transfection, Western Blot, Comparison, Negative Control, Expressing, Positive Control

Journal: Scientific Reports

Article Title: New microRNA-based therapies reveal common targets in paediatric medulloblastoma and adult glioblastoma

doi: 10.1038/s41598-025-05517-9

Figure Lengend Snippet: Transient transfections of GB cells. ( a, b ) miR-206 and miR-383 were significantly upregulated after transfections with miR-206 and miR-383 mimics within both GB cell lines (U251MG miR-206, p < 0.0001; U87MG miR-206, p < 0.0034; U251MG and U87MG miR-383, p < 0.0001). ( c ) CORO1C was significantly downregulated post transfection with miR-206 mimic in U251MG and U87MG cells (U251MG, p < 0.0001; U87MG, P < 0.0001). ( d ) SV2B was significantly downregulated post transfection with miR-383 mimic with both GB cell lines (U251MG, p < 0.0001; U87MG, p < 0.0001). ( e, f ) GB cell viability was significantly decreased post transfection with miR-206 and miR-383 mimics in both, U251MG and U87MG cells (U251MG miR-206, p < 0.0039; U87MG miR-206, p < 0.04; U251MG miR-383, p < 0.002; U87MG miR-383, p < 0.001). ( g, h ) GB cells lost their colony forming abilities post transfections with miR-206 and miR-383 mimics (p < 0.0001). ( i ) Microscopic images of CF within GB cells post exposure to miR-206 and miR-383 mimics (× 40 magnification). ( j ) Western blotting analysis in U251MG showed that the protein levels of CORO1C were significantly decreased in comparison to the negative control condition post exposure to miR-206 mimic (p < 0.0001), whilst the levels of CORO1C were unaffected in the miR-383 mimic condition. The protein levels of SV2B were also significantly reduced in comparison to the negative control condition post exposure to miR-383 mimic (p < 0.0001). Additionally, the levels of SV2B were also downregulated in the miR-206 mimic condition (p < 0.0001). ( k ) Western blotting analysis in U87MG showed that the protein levels of CORO1C were significantly decreased in comparison to the negative control condition post exposure to miR-383 mimic p < 0.0002), whilst the levels of CORO1C were unaffected in the miR-206 mimic condition. The protein levels of SV2B were also significantly reduced in comparison to the negative control condition post exposure to miR-383 mimic(p < 0.0001). Additionally, the levels of SV2B were not affected in the miR-206 mimic condition. ( l, m ) Chemiluminescent images showing the expression of β-actin, CORO1C, and SV2B within U251MG and U87MG cells post exposure to negative control miRNA #1, positive control miR-1, and miR-206 and miR-383 mimics. Results are based on three independent experiments, n = 3. All statistical analyses were conducted using ordinary one-way ANOVA, followed by Dunnett’s multiple comparisons test. Error bars represent the SD of the mean. Original blots/gels are presented in Supplementary Fig. . ns non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Following this, the PVDF membranes were incubated with TBST and the appropriate primary antibody antisera against CORO1C (dilution 1:500), SV2B (dilution 1:500), and β-actin (dilution 1:1000) overnight at 4 °C (ProteinTech, Manchester, UK).

Techniques: Transfection, Western Blot, Comparison, Negative Control, Expressing, Positive Control