Scholarlink

Background <p>Glioblastoma is the commonest malignant brain tumor and has a very poor prognosis. Reduced expression of the <i>MGMT</i> gene (10q26.3), influenced primarily by the methylation of two differentially methylated regions (DMR1 and DMR2), is associated with a good response to temozolomide treatment. However, suitable methods for detecting the methylation of the <i>MGMT</i> gene promoter and setting appropriate cutoff values are debated.</p> Results <p>A cohort of 108 patients with histologically and genetically defined glioblastoma was retrospectively examined with methylation-specific Sanger sequencing (sSeq) and methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) methods. The DMR2 region was methylated in 29% of samples, whereas DMR1 was methylated in 12% of samples. Methylation detected with the MS-MLPA method using probes MGMT_215, MGMT_190, and MGMT_124 from the ME012-A1 kit (located in DMR1 and DMR2) correlated with the methylation of the corresponding CpG dinucleotides detected with sSeq (<i>p</i> = 0.005 for probe MGMT_215; <i>p</i> &lt; 0.001 for probe MGMT_190; <i>p</i> = 0.016 for probe MGMT_124). The threshold for methylation detection with the MS-MLPA method was calculated with a ROC curve analysis and principal components analysis of the data obtained with the MS-MLPA and sSeq methods, yielding a weighted value of 0.362. Thus, methylation of the <i>MGMT</i> gene promoter was confirmed in 36% of samples. These patients had statistically significantly better overall survival (<i>p</i> = 0.003).</p> Conclusions <p>Our results show that the threshold for methylation detection with the MS-MLPA method determined here is useful from a diagnostic perspective because it allows the stratification of patients who will benefit from specific treatment protocols, including temozolomide. Detailed analysis of the <i>MGMT</i> gene promoter enables the more-precise and personalized treatment of patients with glioblastoma.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Validating a clinically based MS-MLPA threshold through comparison with Sanger sequencing in glioblastoma patients

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年1月29日

查看全文
  • Halka Lhotska,
  • Karolina Janeckova,
  • Hana Cechova,
  • Jaromir Macoun,
  • Tatiana Aghova,
  • Libuse Lizcova,
  • Karla Svobodova,
  • Lucie Hodanova,
  • Dora Konecna,
  • Jiri Soukup,
  • Filip Kramar,
  • David Netuka,
  • Zuzana Zemanova

摘要

Background

Glioblastoma is the commonest malignant brain tumor and has a very poor prognosis. Reduced expression of the MGMT gene (10q26.3), influenced primarily by the methylation of two differentially methylated regions (DMR1 and DMR2), is associated with a good response to temozolomide treatment. However, suitable methods for detecting the methylation of the MGMT gene promoter and setting appropriate cutoff values are debated.

Results

A cohort of 108 patients with histologically and genetically defined glioblastoma was retrospectively examined with methylation-specific Sanger sequencing (sSeq) and methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) methods. The DMR2 region was methylated in 29% of samples, whereas DMR1 was methylated in 12% of samples. Methylation detected with the MS-MLPA method using probes MGMT_215, MGMT_190, and MGMT_124 from the ME012-A1 kit (located in DMR1 and DMR2) correlated with the methylation of the corresponding CpG dinucleotides detected with sSeq (p = 0.005 for probe MGMT_215; p < 0.001 for probe MGMT_190; p = 0.016 for probe MGMT_124). The threshold for methylation detection with the MS-MLPA method was calculated with a ROC curve analysis and principal components analysis of the data obtained with the MS-MLPA and sSeq methods, yielding a weighted value of 0.362. Thus, methylation of the MGMT gene promoter was confirmed in 36% of samples. These patients had statistically significantly better overall survival (p = 0.003).

Conclusions

Our results show that the threshold for methylation detection with the MS-MLPA method determined here is useful from a diagnostic perspective because it allows the stratification of patients who will benefit from specific treatment protocols, including temozolomide. Detailed analysis of the MGMT gene promoter enables the more-precise and personalized treatment of patients with glioblastoma.