PURPOSE: The diagnosis of leptomeningeal metastases is usually confirmed by the finding of malignant cells by cytologic examination in the cerebrospinal fluid (CSF). More sensitive and specific cancer biomarkers may improve the detection of tumor cells in the CSF. Promoter methylation of the human telomerase reverse transcriptase (hTERT) gene characterizes most cancer cells. The aim of this study was to develop a sensitive method to detect hTERT methylation and to explore its use as a cancer biomarker in CSF. EXPERIMENTAL DESIGN: In 77 CSF specimens from 67 patients, hTERT promoter methylation was evaluated using real-time methylation-sensitive high-resolution melting (MS-HRM) and real-time TaqMan PCR and MS-HRM in a single-tube assay. RESULTS: Real-time MS-HRM assay was able to detect down to 1% hTERT-methylated DNA in a background of unmethylated DNA. PCR products were obtained from 90% (69/77) of CSF samples. No false positive hTERT was detected in the 21 non-neoplastic control cases, given to the method a specificity of 100%. The sensitivity of the real-time MS-HRM compared with the cytologic gold standard analysis was of 92% (11/12). Twenty-six CSFs from 22 patients with an hTERT-methylated primary tumor showed cytologic results suspicious for malignancy; in 17 (65%) of them, a diagnosis of leptomeningeal metastases could be confirmed by the hTERT methylation test. CONCLUSION: The hTERT real-time MS-HRM approach is fast, specific, sensitive, and could therefore become a valuable tool for diagnosis of leptomeningeal metastases as an adjunct to the traditional examination of CSF.
PURPOSE: The diagnosis of leptomeningeal metastases is usually confirmed by the finding of malignant cells by cytologic examination in the cerebrospinal fluid (CSF). More sensitive and specific cancer biomarkers may improve the detection of tumor cells in the CSF. Promoter methylation of the humantelomerase reverse transcriptase (hTERT) gene characterizes most cancer cells. The aim of this study was to develop a sensitive method to detect hTERT methylation and to explore its use as a cancer biomarker in CSF. EXPERIMENTAL DESIGN: In 77 CSF specimens from 67 patients, hTERT promoter methylation was evaluated using real-time methylation-sensitive high-resolution melting (MS-HRM) and real-time TaqMan PCR and MS-HRM in a single-tube assay. RESULTS: Real-time MS-HRM assay was able to detect down to 1% hTERT-methylated DNA in a background of unmethylated DNA. PCR products were obtained from 90% (69/77) of CSF samples. No false positive hTERT was detected in the 21 non-neoplastic control cases, given to the method a specificity of 100%. The sensitivity of the real-time MS-HRM compared with the cytologic gold standard analysis was of 92% (11/12). Twenty-six CSFs from 22 patients with an hTERT-methylated primary tumor showed cytologic results suspicious for malignancy; in 17 (65%) of them, a diagnosis of leptomeningeal metastases could be confirmed by the hTERT methylation test. CONCLUSION: The hTERT real-time MS-HRM approach is fast, specific, sensitive, and could therefore become a valuable tool for diagnosis of leptomeningeal metastases as an adjunct to the traditional examination of CSF.
Authors: Lindsay Angus; John W M Martens; Martin J van den Bent; Peter A E Sillevis Smitt; Stefan Sleijfer; Agnes Jager Journal: Neuro Oncol Date: 2019-03-18 Impact factor: 12.300
Authors: Yuxuan Wang; Simeon Springer; Ming Zhang; K Wyatt McMahon; Isaac Kinde; Lisa Dobbyn; Janine Ptak; Henry Brem; Kaisorn Chaichana; Gary L Gallia; Ziya L Gokaslan; Mari L Groves; George I Jallo; Michael Lim; Alessandro Olivi; Alfredo Quinones-Hinojosa; Daniele Rigamonti; Greg J Riggins; Daniel M Sciubba; Jon D Weingart; Jean-Paul Wolinsky; Xiaobu Ye; Sueli Mieko Oba-Shinjo; Suely K N Marie; Matthias Holdhoff; Nishant Agrawal; Luis A Diaz; Nickolas Papadopoulos; Kenneth W Kinzler; Bert Vogelstein; Chetan Bettegowda Journal: Proc Natl Acad Sci U S A Date: 2015-07-20 Impact factor: 11.205
Authors: Emilie Le Rhun; Andrew Kramar; Sophie Salingue; Marie Girot; Isabelle Rodrigues; Audrey Mailliez; Fahed Zairi; Edgar Bakhache; Yves Marie Robin; Sophie Taillibert; François Dubois; Jacques Bonneterre; Marc C Chamberlain Journal: J Neurooncol Date: 2014-01-28 Impact factor: 4.130