Rapid detection of IDH mutations in gliomas by intraoperative mass spectrometry

The development and performance of two mass spectrometry (MS) workflows for the intraoperative diagnosis of isocitrate dehydrogenase (IDH) mutations in glioma is implemented by independent teams at Mayo Clinic, Jacksonville, and Huashan Hospital, Shanghai. The infiltrative nature of gliomas makes rapid diagnosis necessary to guide the extent of surgical resection of central nervous system (CNS) tumors. The combination of tissue biopsy and MS analysis used here satisfies this requirement. The key feature of both described methods is the use of tandem MS to measure the oncometabolite 2-hydroxyglutarate (2HG) relative to endogenous glutamate (Glu) to characterize the presence of mutant tumor. The experiments i) provide IDH mutation status for individual patients and ii) demonstrate a strong correlation of 2HG signals with tumor infiltration. The measured ratio of 2HG to Glu correlates with IDH-mutant (IDH-mut) glioma (P < 0.0001) in the tumor core data of both teams. Despite using different ionization methods and different mass spectrometers, comparable performance in determining IDH mutations from core tumor biopsies was achieved with sensitivities, specificities, and accuracies all at 100%. None of the 31 patients at Mayo Clinic or the 74 patients at Huashan Hospital were misclassified when analyzing tumor core biopsies. Robustness of the methodology was evaluated by postoperative re-examination of samples. Both teams noted the presence of high concentrations of 2HG at surgical margins, supporting future use of intraoperative MS to monitor for clean surgical margins. The power of MS diagnostics is shown in resolving contradictory clinical features, e.g., in distinguishing gliosis from IDH-mut glioma.

With the arrival of genomics, molecular diagnostics has grown in importance as a complement to microscopic observations. In brain cancer, some gliomas have mutations in an enzyme associated with the isocitrate biochemical pathway which result in the accumulation of a distinctive metabolite, 2HG. Measurement of brain tissue during surgery using a mass spectrometer (MS) provides information on isocitrate dehydrogenase (IDH)-mutation status not available prior to or during surgery. The data suggest that MS could be used to identify IDH-mut tissue margins at locations of interest selected by the surgeon intraoperatively and rapidly within 2 min. Thus, this molecular measurement should assist the surgeon in achieving optimal surgical resection of infiltrated tissue in those cases where the tumor core is IDH-mut positive.