PURPOSE: Conventional genetic analyzers require surgically obtained tumor tissues to confirm the molecular diagnosis of diffuse glioma. Recent technical breakthroughs have enabled increased utilization of cell-free tumor DNA (ctDNA) in body fluids as a reliable resource for molecular diagnosis in various cancers. Here, we tested the application of a chip-based digital PCR system for the less invasive diagnosis (i.e., liquid biopsy) of diffuse glioma using the cerebrospinal fluid (CSF). METHODS: CSF samples from 34 patients with diffuse glioma were collected from the surgical field during craniotomy. Preoperative lumbar CSF collection was also performed in 11 patients. Extracted ctDNA was used to analyze diagnostic point mutations in IDH1 R132H, TERT promoter (C228T and C250T), and H3F3A (K27M) on the QuantStudio® 3D Digital PCR System. These results were compared with their corresponding tumor DNA samples. RESULTS: We detected either of the diagnostic mutations in tumor DNA samples from 28 of 34 patients. Among them, we achieved precise molecular diagnoses using intracranial CSF in 20 (71%). Univariate analyses revealed that the World Health Organization (WHO) grade (p = 0.0034), radiographic enhancement (p = 0.0006), and Mib1 index (p = 0.01) were significant predictors of precise CSF-based molecular diagnosis. We precisely diagnosed WHO grade III or IV diffuse gliomas using lumbar CSF obtained from 6 (87%) of 7 patients with tumors harboring any mutation. CONCLUSION: We established a novel, non-invasive molecular diagnostic method using a chip-based digital PCR system targeting ctDNA derived from CSF with high sensitivity and specificity, especially for high-grade gliomas.
PURPOSE: Conventional genetic analyzers require surgically obtained tumor tissues to confirm the molecular diagnosis of diffuse glioma. Recent technical breakthroughs have enabled increased utilization of cell-free tumor DNA (ctDNA) in body fluids as a reliable resource for molecular diagnosis in various cancers. Here, we tested the application of a chip-based digital PCR system for the less invasive diagnosis (i.e., liquid biopsy) of diffuse glioma using the cerebrospinal fluid (CSF). METHODS: CSF samples from 34 patients with diffuse glioma were collected from the surgical field during craniotomy. Preoperative lumbar CSF collection was also performed in 11 patients. Extracted ctDNA was used to analyze diagnostic point mutations in IDH1R132H, TERT promoter (C228T and C250T), and H3F3A (K27M) on the QuantStudio® 3D Digital PCR System. These results were compared with their corresponding tumor DNA samples. RESULTS: We detected either of the diagnostic mutations in tumor DNA samples from 28 of 34 patients. Among them, we achieved precise molecular diagnoses using intracranial CSF in 20 (71%). Univariate analyses revealed that the World Health Organization (WHO) grade (p = 0.0034), radiographic enhancement (p = 0.0006), and Mib1 index (p = 0.01) were significant predictors of precise CSF-based molecular diagnosis. We precisely diagnosed WHO grade III or IV diffuse gliomas using lumbar CSF obtained from 6 (87%) of 7 patients with tumors harboring any mutation. CONCLUSION: We established a novel, non-invasive molecular diagnostic method using a chip-based digital PCR system targeting ctDNA derived from CSF with high sensitivity and specificity, especially for high-grade gliomas.
Authors: Tareq A Juratli; Sebastian Stasik; Dietmar Krex; Christian Thiede; Amir Zolal; Caroline Schuster; Sven Richter; Dirk Daubner; Mazen A Juratli; Rachel Thowe; Silke Hennig; Meriem Makina; Matthias Meinhardt; Tim Lautenschlaeger; Gabriele Schackert Journal: Clin Cancer Res Date: 2018-06-25 Impact factor: 12.531
Authors: David N Louis; Pieter Wesseling; Werner Paulus; Caterina Giannini; Tracy T Batchelor; J Gregory Cairncross; David Capper; Dominique Figarella-Branger; M Beatriz Lopes; Wolfgang Wick; Martin van den Bent Journal: Acta Neuropathol Date: 2018-01-25 Impact factor: 17.088
Authors: Daniel J Brat; Kenneth Aldape; Howard Colman; Eric C Holland; David N Louis; Robert B Jenkins; B K Kleinschmidt-DeMasters; Arie Perry; Guido Reifenberger; Roger Stupp; Andreas von Deimling; Michael Weller Journal: Acta Neuropathol Date: 2018-09-26 Impact factor: 17.088
Authors: Melike Pekmezci; Terri Rice; Annette M Molinaro; Kyle M Walsh; Paul A Decker; Helen Hansen; Hugues Sicotte; Thomas M Kollmeyer; Lucie S McCoy; Gobinda Sarkar; Arie Perry; Caterina Giannini; Tarik Tihan; Mitchel S Berger; Joseph L Wiemels; Paige M Bracci; Jeanette E Eckel-Passow; Daniel H Lachance; Jennifer Clarke; Jennie W Taylor; Tracy Luks; John K Wiencke; Robert B Jenkins; Margaret R Wrensch Journal: Acta Neuropathol Date: 2017-03-02 Impact factor: 15.887
Authors: Florent Mouliere; Richard Mair; Dineika Chandrananda; Francesco Marass; Christopher G Smith; Jing Su; James Morris; Colin Watts; Kevin M Brindle; Nitzan Rosenfeld Journal: EMBO Mol Med Date: 2018-12 Impact factor: 12.137
Authors: Riccardo Soffietti; Chetan Bettegowda; Ingo K Mellinghoff; Katherine E Warren; Manmeet S Ahluwalia; John F De Groot; Evanthia Galanis; Mark R Gilbert; Kurt A Jaeckle; Emilie Le Rhun; Roberta Rudà; Joan Seoane; Niklas Thon; Yoshie Umemura; Michael Weller; Martin J van den Bent; Michael A Vogelbaum; Susan M Chang; Patrick Y Wen Journal: Neuro Oncol Date: 2022-06-01 Impact factor: 13.029