Journal: Journal of Neuro-Oncology

Article Title: MGMT promoter methylation status for glioblastoma: defining the clinically relevant cut-off value for pyrosequencing

doi: 10.1007/s11060-026-05633-0

Figure Lengend Snippet: Flowchart of patient selection. MGMTpm, MGMT promoter methylation; RT, radiotherapy; TMZ, Temozolomide. * Meningioma, ependymoma, diffuse midline glioma, pilocytic astrocytoma, ganglioglioma, fibrillary astrocytoma, oligodendroglioma IDHmut, gemistocytic astrocytoma IDHmut and germinoma ** Including patients not receiving postoperative treatment. *** preopPS, or extent of resection **** thereby absent from the SQR

Article Snippet: A third option is to establish a cut-off based on outcome, such as in the study by Quillien where the Pyromark MGMT kit investigating CpGs 74–78 in exon 1 of the MGMT gene by pyrosequencing was used and a cut-off of ≥ 9% for mMGMT was proposed [ ].

Techniques: Selection, Methylation

Journal: Journal of Neuro-Oncology

Article Title: MGMT promoter methylation status for glioblastoma: defining the clinically relevant cut-off value for pyrosequencing

doi: 10.1007/s11060-026-05633-0

Figure Lengend Snippet: MGMT promoter methylation status and cut-offs. ( a ) Distribution of mean MGMTpm status of all 451 patients. The solid green line representing the supervised cut-off ( b ) and the dashed black line representing the unsupervised cut-off ( c ). Distributions for each individual CpG are presented in Supplementary Fig. . ( b ) Cut-off supervised by OS and adjusted for prognostic factors: age, extent of resection and preoperative ECOG performance status (solid green line), determined through 5-fold cross-validation with the highest concordance index corresponding to the best performing AFT (accelerated failure time) model. ( c ) Bimodal normal mixture model, with unsupervised cut-off (dashed black line) determined as the intersection point of the two distributions. Blue bars represent patients with methylated MGMT and red bars represent patients with unmethylated MGMT, according to the supervised cut-off (solid green line, b )

Article Snippet: A third option is to establish a cut-off based on outcome, such as in the study by Quillien where the Pyromark MGMT kit investigating CpGs 74–78 in exon 1 of the MGMT gene by pyrosequencing was used and a cut-off of ≥ 9% for mMGMT was proposed [ ].

Techniques: Methylation, Biomarker Discovery

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Progesterone boosts abiraterone-driven target and NK cell therapies against glioblastoma

doi: 10.1186/s13046-024-03144-2

Figure Lengend Snippet: Effect of Prog on MGMT expression. A , B After treatment with Prog in the absence and presence of Abi, T98G cells were harvested for Western blotting (Left) and RT-qPCR (Right). C Left: After treatment for 48 h, T98G cells were treated with 20 µM cycloheximide (CHX) for the indicated time interval, and cell lysates were collected for Western blotting. Right: Quantitative results. D After treatment with Prog, gDNA of T98G and U87MG-R cells F was purified, and subjected to pyrosequencing analysis. The percentage of methylation in 5 CpG residues in T98G and U87MG-R cells were quantified. Experiments were performed independently three times. (* P < 0.05, ** P < 0.01)

Article Snippet: CpGs methylation are tested with the PyroMark Q24 CpG MGMT kit provided by Qiagen [ , ].

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Purification, Methylation

Journal: Cancers

Article Title: ‘The Reports of My Death Are Greatly Exaggerated’—Evaluating the Effect of Necrosis on MGMT Promoter Methylation Testing in High-Grade Glioma

doi: 10.3390/cancers16101906

Figure Lengend Snippet: Overview of intratumoral cellularity, necrosis and MGMT p methylation percentages across 52 primary HGG cases with repeated MGMT p testing. Horizontal red lines indicate the borderline MGMT p zone (9–13%).

Article Snippet: Pyrosequencing of the MGMT p region was undertaken using a PyroMark therascreen MGMT kit (Qiagen) and the PyroMark Q24 system (Qiagen), which detect four CpG sites located in exon 1.

Techniques: Methylation

Journal: Cancers

Article Title: ‘The Reports of My Death Are Greatly Exaggerated’—Evaluating the Effect of Necrosis on MGMT Promoter Methylation Testing in High-Grade Glioma

doi: 10.3390/cancers16101906

Figure Lengend Snippet: Summary of primary and recurrent presentations of HGGs with maintenance of MGMT p status.

Article Snippet: Pyrosequencing of the MGMT p region was undertaken using a PyroMark therascreen MGMT kit (Qiagen) and the PyroMark Q24 system (Qiagen), which detect four CpG sites located in exon 1.

Techniques: Blocking Assay, Methylation

Journal: Cancers

Article Title: ‘The Reports of My Death Are Greatly Exaggerated’—Evaluating the Effect of Necrosis on MGMT Promoter Methylation Testing in High-Grade Glioma

doi: 10.3390/cancers16101906

Figure Lengend Snippet: Overview of intratumoral cellularity, necrosis and MGMT p methylation percentages in 23 cases of recurrent HGG. Horizontal red lines indicate the borderline MGMT p zone (9–13%).

Article Snippet: Pyrosequencing of the MGMT p region was undertaken using a PyroMark therascreen MGMT kit (Qiagen) and the PyroMark Q24 system (Qiagen), which detect four CpG sites located in exon 1.

Techniques: Methylation

Journal: Cancers

Article Title: ‘The Reports of My Death Are Greatly Exaggerated’—Evaluating the Effect of Necrosis on MGMT Promoter Methylation Testing in High-Grade Glioma

doi: 10.3390/cancers16101906

Figure Lengend Snippet: Correlation analyses demonstrate the reproducibility of MGMT p methylation testing in HGG sections with varying degrees of tumor necrosis. Repeated measures correlation analysis shows no significant association between MGMT p status and tumor cellularity ( A ) or necrosis ( B ). Each participant, plotted in a different color, contributes two paired data points representing repeated MGMT p tests on different tissue blocks. The corresponding lines depict the repeated measures correlation model. The relationship of (black) line-of-best-fit between MGMT p methylation percentages (data points depicted by red triangles) at thresholds of 30% necrosis and 60% tumor cellularity was determined by Spearman rank correlation ( C ).

Article Snippet: Pyrosequencing of the MGMT p region was undertaken using a PyroMark therascreen MGMT kit (Qiagen) and the PyroMark Q24 system (Qiagen), which detect four CpG sites located in exon 1.

Techniques: Methylation

Journal: Cancers

Article Title: ‘The Reports of My Death Are Greatly Exaggerated’—Evaluating the Effect of Necrosis on MGMT Promoter Methylation Testing in High-Grade Glioma

doi: 10.3390/cancers16101906

Figure Lengend Snippet: Changes in MGMT p status by degree of necrosis in NT block.

Article Snippet: Pyrosequencing of the MGMT p region was undertaken using a PyroMark therascreen MGMT kit (Qiagen) and the PyroMark Q24 system (Qiagen), which detect four CpG sites located in exon 1.

Techniques: Blocking Assay

Journal: Cancers

Article Title: ‘The Reports of My Death Are Greatly Exaggerated’—Evaluating the Effect of Necrosis on MGMT Promoter Methylation Testing in High-Grade Glioma

doi: 10.3390/cancers16101906

Figure Lengend Snippet: Overview of tumoral cellularity, necrosis and MGMT p methylation percentages in 19 cases of recurrent HGG (16 recurrences; 3 re-resected recurrences). Horizontal red lines indicate the borderline MGMT p zone (9–13%).

Article Snippet: Pyrosequencing of the MGMT p region was undertaken using a PyroMark therascreen MGMT kit (Qiagen) and the PyroMark Q24 system (Qiagen), which detect four CpG sites located in exon 1.

Techniques: Methylation