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GENE-41. LIQUID BIOPSY USING CELL FREE DNA FROM THE CEREBROSPINAL FLUID (CSF) IN GLIOMA

GENE-41. LIQUID BIOPSY USING CELL FREE DNA FROM THE CEREBROSPINAL FLUID (CSF) IN GLIOMA

Authors :
Tomonari Suzuki
Ryo Nishikawa
Kazuhiko Mishima
Jun-ichi Adachi
Publication Year :
2018
Publisher :
Oxford University Press, 2018.

Abstract

Mutation in the IDH1gene is a genetic defect exclusively found in glioma, and has been established as a prognostic factor for WHO Grade 2–4 glioma. Therefore, detecting this mutation is clinically important. In this study, we report the results of analyzing defects in IDH1 mutation by detecting circulating cell free (ccf) DNA in the CSF derived from the tumor tissue of glioma patients. Moreover, MGMTpromoter methylation, H3F3A and BRAFmutation were evaluated for the CSF samples in a subset of glioma patients. Lumbar puncture was performed to obtain CSF from 7 patients with glioma. ccfDNA was extracted from 1 ml of CSF using the Maxwell rapid sample concentrator system. Subsequently, the presence of point mutation of theIDH1gene was screened by real-time PCR/high-resolution melting (HRM) curve analysis, and the mutation was confirmed on the basis of DNA sequencing results. Status in the IDH1gene was also analyzed using the same assay technique for the DNA extracted from the excised fresh tumor tissue, to compare the results with that of CSF-derived ccfDNA. In addition, ccfDNA was also extracted from the plasma in 3 patients to additionally examine the presence of the IDH1gene mutation. MGMTpromoter methylation, H3F3A and BRAFmutation were also performed using the HRM method. CSF-derived ccfDNA was successfully extracted from all patients and analyzed. IDH1gene mutation was detected in 3 of the 7 glioma patients. The results of the IDH1gene analysis of CSF-derived ccfDNA and that of the DNA extracted from the surgically excised tumor tissue were consisted in all patients. Three MGMTpromoter methylation, two H3F3Amutations, and three BRAFmutations were detected, respectively. Gene analysis of ccfDNA from the CSF enabled the evaluation of IDH1 mutation in glioma patients less invasively, without directly obtaining any tumor tissue. Moreover, this technique can be applied to analyze MGMTgene promotor methylation, H3F3A and BRAFmutation.

Details

Language :
English
Database :
OpenAIRE
Accession number :
edsair.doi.dedup.....5df64f72824af9e4d6c302fadc6df711