scholarly journals PATH-08. CHALLENGES IN OPTIMIZATION OF MGMT PROMOTER METHYLATION ASSAYS FOR DEVELOPMENT OF COMPANION DIAGNOSIS IN GLIOBLASTOMA

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi144-vi144
Author(s):  
Motoo Nagane ◽  
Kuniaki Saito ◽  
Keiichi Kobayashi ◽  
Saki Shimizu ◽  
Nobuyoshi Sasaki ◽  
...  
Keyword(s):  
Methylation Status ◽  
Mgmt Promoter Methylation ◽  
Targeted Drugs ◽  
Newly Diagnosed ◽  
Companion Diagnostics ◽  
Bisulfite Treatment ◽  
Cpg Sites ◽  
Multiple Issues ◽  
High Concordance

Abstract BACKGROUNDS Despite recent limelight of precision medicine in oncology, effective targeted drugs and diagnostics for molecular markers have not been developed or approved for glioblastoma (GBM) yet. Notably, there remain multiple issues in MGMT testing, including determination of optimal assays and cutoff for methylation status, and its value as “companion diagnostics.” We compared two of MGMT assays, qualitative standard MSP and pyrosequencing, in light of compatibility, predictability, and cutoff points. METHODS Sixty-five newly diagnosed GBMs with two MGMT assays performed since 2006. Survival was analyzed for 48 GBMs with wildtype IDH and initial temozolomide use. Extracted DNA was subjected to bisulfite treatment, thereby MGMT status was determined. RESULTS Among 65 GBMs, MGMT was methylated (M) in 34, and unmethylated (U) in 31, by MSP. Median methylation rate at CpG sites by Pyro was 15.9%, with average 23.0% for M and 9.5% for U (Mann-Whitney-U, P< 0.001). MGMT status by MSP significantly correlated with that by Pyro with cutoff between 10 – 16% (Fisher, P< 0.001). Patients with MGMT M by MSP showed significantly longer PFS and OS than with U (PFS 13.5 m vs. 5.6 m, P = 0.005; OS 37.7 m vs. 15.8 m, P = 0.001). In contrast, by Pyro (cutoff 14%), only PFS was significantly better in M (11.7 m vs 5.6 m, P = 0.014), but not OS (27.9 m vs. 15.8 m, P = 0.249). No substantial gain for PFS was noted in MGMT M GBMs beyond cutoff 12% by Pyro. Multivariate analysis revealed MGMT M and KPS 70 or higher as independent factors for better PFS and OS using MSP, but only for PFS with Pyro. CONCLUSIONS In this study, qualitative MSP exhibited rather better discrimination of prognosis than quantitative Pyro, although showing a high concordance for MGMT status, necessitating further confirmation among assays.

P04.22 Tumor treating fields in high-grade glioma patients: A retrospective single-center study

2021 ◽  
Vol 23 (Supplement_2) ◽  
pp. ii23-ii23
Author(s):  
R Lucaciu ◽  
B Suchorska ◽  
M Wettig ◽  
S Jung ◽  
M Scholz
Keyword(s):  
Promoter Methylation ◽  
Daily Life ◽  
Methylation Status ◽  
Mgmt Promoter Methylation ◽  
Therapeutic Modality ◽  
High Grade ◽  
Newly Diagnosed ◽  
Non Invasive ◽  
Tumor Treating Fields ◽  
Mgmt Promoter

Abstract BACKGROUND Tumor-treating fields (TTFields) are a modern anti-mitotic, non-invasive therapy for the treatment of patients with recurrent and newly diagnosed glioblastoma multiforme (GBM). In Europe, Optune® recieved in 2015 the CE certification. TTFields are a low-intensity (1–3 V/cm) approved therapeutic modality using a non-invasive application of intermediate frequency (200 kHz) alternating electric fields through four transducer arrays directly applied to the skin. The EF-14 study has shown that the addition of TTFields to temozolomide chemotherapy in patients with newly diagnosed GBM significantly improved overall survival (OS) and progression-free survival (PFS) without additional adverse events, apart from mild to moderate skin irritations (Stupp et al., JAMA 2017). MATERIAL We retrospectively analyzed data from TTFields-treated patients (2015–2020) that were treated at our department. Patient characteristics such as MGMT promoter methylation status, age, and diagnosis, as well as treatment duration and TTFields therapy usage, were evaluated for this study. RESULTS 29 patients were treated with TTFields therapy between 2015 and 2020 at our hospital. Most patients received TTFields as primary treatment together with temozolomide maintenance therapy. In detail, 48% of patients were diagnosed with newly diagnosed GBM, 41% received TTFields therapy after tumor recurrence and 10% were diagnosed with other high-grade gliomas. In summary, patients could integrate TTFields therapy into their daily life and showed high adherence to the therapy.Particularly, one of our patients (with MGMT-promoter methylation positive) receives TTFields therapy now for almost 1229 days (approx. 41 months) and is still on therapy. Additionally, this patient shows a high usage rate of 86% indicating well integration of the therapy into daily life. CONCLUSION Taken together, our data provided the outcomes of using TTFields together with chemotherapy in the treatment of recurrent and newly diagnosed GBM in our department. Therapy with TTFields has been showing to provide significant clinical benefit for GBM patients.

Correlation of the quantitative level of MGMT promoter methylation and overall survival in primary diagnosed glioblastomas using the quantitative MethyQESD method

2019 ◽  
Vol 73 (2) ◽  
pp. 112-115 ◽  
Author(s):  
Charlotte von Rosenstiel ◽  
Benedikt Wiestler ◽  
Bernhard Haller ◽  
Friederike Schmidt-Graf ◽  
Jens Gempt ◽  
...  
Keyword(s):  
Overall Survival ◽  
Promoter Methylation ◽  
Promoter Region ◽  
Dna Methyltransferase ◽  
Methylation Status ◽  
Mgmt Promoter Methylation ◽  
Newly Diagnosed ◽  
Quantitative Level ◽  
Mgmt Promoter ◽  
The Impact

AimsO(6)-methylguanine-DNA-methyltransferase (MGMT) promoter methylation is a high predictive factor for therapy results of temozolomide in patients with glioma. The objective of this work was to analyse the impact of MGMT promoter methylation in patients with primary diagnosed glioblastoma (GBM) relating to survival using a quantitative method (methylation quantification of endonuclease-resistant DNA, MethyQESD) by verifying a cut-off point for MGMT methylation provided by the literature (</≥10%) and calculating an optimal cut-off.Methods67 patients aged 70 years or younger, operated between January 2013 and December 2015, with newly diagnosed IDH wild-type GBM and clinical follow-up were retrospectively investigated in this study. A known MGMT promoter methylation status was the inclusion criteria.ResultsMedian overall survival (OS) was 16.9 months. Patients who had a methylated MGMT promoter region of ≥10% had an improved OS compared with patients with a methylated promoter region of <10% (p=0.002). Optimal cut-off point for MGMT promoter methylation was 11.7% (p=0.012).ConclusionThe results confirm that the quantitative level of MGMT promoter methylation is a positive prognostic factor in newly diagnosed patients with GBM. The cut-off provided by the literature (</≥10%) and the calculated optimal cut-off value of 11.7% give a statistically significant separation. Hence, MethyQESD is a reliable method to calculate MGMT promoter methylation in GBM.

The clinical significance of telomerase reverse transcriptase (TERT) promoter mutations, telomere length and O6-methylguanine DNA methyltransferase (MGMT) promoter methylation status in newly diagnosed and recurrent IDH-wildtype glioblastoma (GBM) patients (PTS): A large mono-institutional study.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. 2053-2053
Author(s):  
Giuseppe Lombardi ◽  
Silvia Giunco ◽  
Francesco Cavallin ◽  
Chiara Angelini ◽  
Mario Caccese ◽  
...  
Keyword(s):  
Telomere Length ◽  
Methylation Status ◽  
Mgmt Promoter Methylation ◽  
Newly Diagnosed ◽  
Mgmt Methylation ◽  
Tert Promoter ◽  
Tert Promoter Mutations ◽  
Mgmt Promoter ◽  
Promoter Mutations ◽  
Mgmt Promoter Methylation Status

2053 Background: the clinical significance of TERT promoter mutations, telomere length and their interactions with MGMT promoter methylation status in patients with IDH-wildtype GBM patients remain unclear. We performed a large mono-institutional study to better investigate their impact and their interaction on clinical outcomes Methods: TERT promoter mutations (C228T and C250T), relative telomere length (RTL) and MGMT methylation status were assessed in 278 newly diagnosed and in 65 recurrent IDH-wildtype GBM PTS which were treated at Veneto Institute of Oncology (Padua, Italy) from Dec 2016 to Jan 2020. We have retrospectively explored association between gene characteristics and neuroradiological response (RANO criteria), progression-free survival (PFS), overall survival (OS). Telomere length was measured by monochrome multiplex PCR and RTL values were calculated as a telomere/single-copy gene ratio Results: characteristics of newly diagnosed GBM PTS were: median age 63 ys, ECOG PS 0-1 in 71% of PTS, radical surgery in 38%, 78% received radiation therapy plus TMZ, MGMT was methylated in 53%, TERT promoter was mutated in 80% (75% C228T, 25% C250T), median RTL was 1.57 (range 0.4-11.37). Objective response rate was reported in 15% of PTS, median OS was 15ms (95% CI 13-18ms), median PFS was 8ms (95% CI 7-9ms). At multivariable analysis, TERT promoter mutations and RTL were not associated with clinical outcomes; about OS, TERT promoter mutations and RTL reported a HR of 1.05 (95% CI 0.64-1.64) and 0.99 (95% CI 0.89-1.10), respectively; MGMT methylated tumors showed significant improved PFS and OS with a HR of 0.54 (95% CI 0.40-0.71) and 0.47 (95% CI 0.34-0.64), respectively. All interactions among MGMT status, TERT mutation status and RTL were not statistically significant. Characteristics of recurrent GBM PTS were: median age 55 ys, ECOG PS 0-1 in 60% of PTS, MGMTmet in 37%, TERT promoter mutations in 75% (75% C228T, 25% C250T), RTL was 1.67 (range 0.68-8.87). At multivariable analysis, only MGMT methylated tumors resulted significantly associated to prolonged OS (HR 0.16; 95% CI 0.07-0.40). No gene interaction was significant. Conclusions: for the first time worldwide, we analyzed the impact of TERT promoter mutations, RTL and MGMT methylation status in both newly diagnosed and recurrent IDH-wildtype GBM PTS. TERT promoter status and RTL were not associated with clinical outcomes at both diagnosis and relapse. MGMT promoter methylation status was the only prognostic factor in both cases. No significant interaction was demonstrated between TERT promoter mutations, RTL and MGMT methylation status.

Hyper-Methylation DAZAP2 May Suppress Its Expression in Specific Subtypes of Myeloma.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4421-4421
Author(s):  
Wei-Xin Hu ◽  
Qiang Qu ◽  
Jiang Li ◽  
Wei Ren
Keyword(s):  
Transcriptional Activity ◽  
Plasma Cells ◽  
Conflicts Of Interest ◽  
Cpg Island ◽  
Methylation Status ◽  
Cpg Islands ◽  
Myeloma Cell ◽  
Positive Control ◽  
Newly Diagnosed ◽  
Cpg Sites

Abstract Abstract 4421 Most malignant features of cancer cells are triggered by activated oncogenes and the loss of tumor suppressors due to mutation or epigenetic inactivation. We previously reported that DAZAP2 was down-regulated in newly diagnosed myeloma (MM) and this may influence MM cell growth and survival. This study was undertaken to evaluate the mechanism of down-regulation DAZAP2 and determine the methylation status and loci of its promoter by using bisulphite genomic-sequencing (BGS) method in KM3 myeloma cell line. Two islands with rich GC box in the promoter of DAZAP2 gene were identified and amplified by using bisulfite-sequencing PCR (BSP). Island 1 spans -472 to -247 including 9 CpG sites (CpGs) and island 2 covers -226 to -13 including 23 CpG sites. The ratio of methylated CpGs in two CpG islands was 46.25%. The above sequences were demethylated and inserted into a pGL2-basic vector. COS-7 cells were transfected with recombinant plasmids and the activity of luciferase was evaluated. The results showed that the CpG island 1 had a weakly transcriptional activity, whereas the CpG island 2 had a strong transcriptional activity (2.38 folds compared with the positive control). The other sequence which covered CpG island 1 and 2 showed a remarkable activity (15.1 folds compared with the positive control). These data indicated that CpG island 2 of DAZAP2 gene may be hypermethylated and suppressed its expression in MM cells. We further correlated DAZAP2 expression with normal plasma cells and malignant myeloma cells, as well as the molecular subtypes which the dataset includes 8 genetic subtypes (MY, PR, LB, MS, HP, CD-1, CD-2, and MF) from 351 newly diagnosed myeloma cases (Zhan, 2006). Interestingly, we extended and confirmed our previous discovery that DAZAP2 was significantly down-regulated in MM cells by using a large uniform dataset (P = 0.004). The low expression of DAZAP2 was especially significant in the subgroups of MY, PR, LB, HP, and CD-1. This study warrants further investigation of DAZAP2 and its potential role in myeloma. Disclosures: No relevant conflicts of interest to declare.

scholarly journals RARE-39. COMBINATION OF TUMOR TREATING FIELDS (TTFIELDS) WITH LOMUSTINE (CCNU) AND TEMOZOLOMIDE (TMZ) IN NEWLY DIAGNOSED GLIOBLASTOMA (GBM) PATIENTS - A BI-CENTRIC ANALYSIS

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi229-vi230
Author(s):  
Lazaros Lazaridis ◽  
Niklas Schäfer ◽  
Teresa Schmidt ◽  
Johannes Weller ◽  
Theophilos-Dimitrios Tzaridis ◽  
...  
Keyword(s):  
Promoter Methylation ◽  
Methylation Status ◽  
Case Series ◽  
Mgmt Promoter Methylation ◽  
Low Grade ◽  
Newly Diagnosed ◽  
Skin Reactions ◽  
Term Survival ◽  
Mgmt Promoter ◽  
Newly Diagnosed Glioblastoma

Abstract BACKGROUND TTFields combined with TMZ chemotherapy demonstrated significantly improved PFS, OS and long-term survival in newly diagnosed glioblastoma (ndGBM) patients in the EF-14 trial; independent of MGMT-promoter methylation-status, age, grade of resection, and performance status. Recently, improved efficacy of lomustine/TMZ compared to TMZ monotherapy in ndGBM patients with MGMT-promotor methylation was reported in the CeTeG trial. As TTFields demonstrated a favorable safety profile as well as a high potential for being combined with other modalities and the encouraging results for methylated MGMT-promoter GBM patients in the CeTeG trial, there is a strong rationale for combining these treatment regimens. Here, we present a case series of patients receiving a combination of TTFields and lomustine/TMZ. METHODS Patients with ndGBM and MGMT-promoter methylation underwent combined therapy of TTFields plus lomustine/TMZ after surgery and radiochemotherapy. MGMT-promoter status was measured by pyrosequencing. Safety, feasibility, and first efficacy results are reported at data cut-off (April 26, 2019). RESULTS Sixteen patients with MGMT-promoter methylated ndGBM (median, range: age 50, 27–70; KPS 90, 60–100) have been treated with a combination of TTFields plus lomustine/TMZ. The analysis included patients with complete resection (n=7), partial resection (n=8), as well as biopsy (n=1). CTCAE grade 3 hematotoxicity was observed in seven patients (44%) but was unlikely related to the addition of TTFields to lomustine/TMZ. Medical device site reactions (low-grade skin reactions) were detected in six patients (38%). At data cut-off, the analyzed patient population demonstrated a median PFS of 20 months; the median OS was not yet reached. CONCLUSION The results of this analysis indicate that the combination of TTFields/lomustine/TMZ is safe and feasible. Moreover, the observed survival outcomes point to preliminary beneficial effects of the triple combination. Additional follow-up and increased sample size are required and planned for further safety and efficacy assessment of this treatment regimen.

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