Clint M. Alfaro, Valentina Pirro, Michael F. Keating, Eyas M. Hattab, R. Graham Cooks and Aaron A. Cohen-Gadol
The authors describe a rapid intraoperative ambient ionization mass spectrometry (MS) method for determining isocitrate dehydrogenase (IDH) mutation status from glioma tissue biopsies. This method offers new glioma management options and may impact extent of resection goals. Assessment of the IDH mutation is key for accurate glioma diagnosis, particularly for differentiating diffuse glioma from other neoplastic and reactive inflammatory conditions, a challenge for the standard intraoperative diagnostic consultation that relies solely on morphology.
Banked glioma specimens (n = 37) were analyzed by desorption electrospray ionization–MS (DESI-MS) to develop a diagnostic method to detect the known altered oncometabolite in IDH-mutant gliomas, 2-hydroxyglutarate (2HG). The method was used intraoperatively to analyze tissue smears obtained from glioma patients undergoing resection and to rapidly diagnose IDH mutation status (< 5 minutes). Fifty-one tumor core biopsies from 25 patients (14 wild type [WT] and 11 mutant) were examined and data were analyzed using analysis of variance and receiver operating characteristic curve analysis.
The optimized DESI-MS method discriminated between IDH-WT and IDH-mutant gliomas, with an average sensitivity and specificity of 100%. The average normalized DESI-MS 2HG signal was an order of magnitude higher in IDH-mutant glioma than in IDH-WT glioma. The DESI 2HG signal intensities correlated with independently measured 2HG concentrations (R2 = 0.98). In 1 case, an IDH1 R132H–mutant glioma was misdiagnosed as a demyelinating condition by frozen section histology during the intraoperative consultation, and no resection was performed pending the final pathology report. A second craniotomy and tumor resection was performed after the final pathology provided a diagnosis most consistent with an IDH-mutant glioblastoma. During the second craniotomy, high levels of 2HG in the tumor core biopsies were detected.
This study demonstrates the capability to differentiate rapidly between IDH-mutant gliomas and IDH-WT conditions by DESI-MS during tumor resection. DESI-MS analysis of tissue smears is simple and can be easily integrated into the standard intraoperative pathology consultation. This approach may aid in solving differential diagnosis problems associated with low-grade gliomas and could influence intraoperative decisions regarding extent of resection, ultimately improving patient outcome. Research is ongoing to expand the patient cohort, systematically validate the DESI-MS method, and investigate the relationships between 2HG and tumor heterogeneity.
Roberto Jose Diaz, Roberto Rey Dios, Eyas M. Hattab, Kelly Burrell, Patricia Rakopoulos, Nesrin Sabha, Cynthia Hawkins, Gelareh Zadeh, James T. Rutka and Aaron A. Cohen-Gadol
Intravenous fluorescein sodium has been used during resection of high-grade gliomas to help the surgeon visualize tumor margins. Several studies have reported improved rates of gross-total resection (GTR) using high doses of fluorescein sodium under white light. The recent introduction of a fluorescein-specific camera that allows for high-quality intraoperative imaging and use of very low dose fluorescein has drawn new attention to this fluorophore. However, the ability of fluorescein to specifically stain glioma cells is not yet well understood.
The authors designed an in vitro model to assess fluorescein uptake in normal human astrocytes and U251 malignant glioma cells. An in vivo experiment was also subsequently designed to study fluorescein uptake by intracranial U87 malignant glioma xenografts in male nonobese diabetic/severe combined immunodeficient mice. A genetically induced mouse glioma model was used to adjust for the possible confounding effect of an inflammatory response in the xenograft model. To assess the intraoperative application of this technology, the authors prospectively enrolled 12 patients who underwent fluorescein-guided resection of their high-grade gliomas using low-dose intravenous fluorescein and a microscope-integrated fluorescence module. Intraoperative fluorescent and nonfluorescent specimens at the tumor margins were randomly analyzed for histopathological correlation.
The in vitro and in vivo models suggest that fluorescein demarcation of glioma-invaded brain is the result of distribution of fluorescein into the extracellular space, most likely as a result of an abnormal blood-brain barrier. Glioblastoma tumor cell–specific uptake of fluorescein was not observed, and tumor cells appeared to mostly exclude fluorescein. For the 12 patients who underwent resection of their high-grade gliomas, the histopathological analysis of the resected specimens at the tumor margin confirmed the intraoperative fluorescent findings. Fluorescein fluorescence was highly specific (up to 90.9%) while its sensitivity was 82.2%. False negatives occurred due to lack of fluorescence in areas of diffuse, low-density cellular infiltration. Margins of contrast enhancement based on intraoperative MRI–guided StealthStation neuronavigation correlated well with fluorescent tumor margins. GTR of the contrast-enhancing area as guided by the fluorescent signal was achieved in 100% of cases based on postoperative MRI.
Fluorescein sodium does not appear to selectively accumulate in astrocytoma cells but in extracellular tumor cell-rich locations, suggesting that fluorescein is a marker for areas of compromised blood-brain barrier within high-grade astrocytoma. Fluorescein fluorescence appears to correlate intraoperatively with the areas of MR enhancement, thus representing a practical tool to help the surgeon achieve GTR of the enhancing tumor regions.
R. Shane Tubbs, Anand N. Bosmia, Marios Loukas, Eyas M. Hattab and Aaron A. Cohen-Gadol
Although it is often visualized surgically, details regarding the inferior medullary velum are lacking in the literature. The present study is intended to better elucidate this neuroanatomical structure using microsurgical and immunohistochemical analyses.
To study the inferior medullary velum, the authors performed microdissection in 15 adult cadavers. Following gross study, specimens were examined histologically.
The inferior medullary velum extended from the flocculus to the middle cerebellar peduncle and stretched between the inferior cerebellar peduncle and the nodule and pyramid. The average thickness of the velum was found to be 0.5 mm (range 0.35–0.8 mm) and the average length was found to be 6 mm (range 5.5–7.2 mm). Arterial branches were identified in all specimens that arose from medullary branches of the posterior inferior cerebellar artery and supplied the inferior medullary velum. Histologically and from internal to external, a choroid plexus epithelium as a single cell layer was adjacent to a cuboidal layer of ependymal cells with no visible cilia. The next layer contained scattered glia in single cells or small clusters. The most external layer was composed of flat spindle cells resembling fibroblasts. No neurons of any type were identified. Only rare axons traversed the thin hypocellular zone that disappeared toward the midline.
Based on this cadaveric study, the authors conclude that division of the inferior medullary velum should be relatively harmless as no neuronal cells were identified in this structure, which appears to be a vestigial bridge of tissue between the left and right sides of the cerebellum.
R. Shane Tubbs, Eyas M. Hattab, Marios Loukas, Joshua J. Chern, Melissa Wellons, John C. Wellons III, Bermans J. Iskandar and Aaron A. Cohen-Gadol
Endocrine dysfunction following endoscopic third ventriculostomy (ETV) is rare, but it has been reported. In the present study the authors sought to determine the histological nature of the floor of the third ventricle in hydrocephalic brains to better elucidate this potential association.
Five adult cadaveric brains with hydrocephalus were examined. Specifically, the floors of the third ventricle of these specimens were studied histologically. Age-matched controls without hydrocephalus were used for comparison.
Although it was thinned in the hydrocephalic brains, the floor of the third ventricle had no significant difference between the numbers of neuronal cell bodies versus nonhydrocephalic brains.
Although uncommon following ETV, endocrine dysfunction has been reported. Based on the present study, this is most likely to be due to the injury of normal neuronal cell bodies found in this location, even in very thinned-out tissue.
R. Shane Tubbs, Mohammadali M. Shoja, Marios Loukas, Jeffrey Lancaster, Martin M. Mortazavi, Eyas M. Hattab and Aaron A. Cohen-Gadol
There is conflicting and often anecdotal evidence regarding the potential motor innervation of the trapezius muscle by cervical nerves, with most authors attributing such fibers to proprioception. As knowledge of such potential motor innervations may prove useful to the neurosurgeon, the present study aimed to elucidate this anatomy further.
Fifteen adult cadavers (30 sides) underwent dissection of the posterior triangle of the neck and harvesting of cervical nerve fibers found to enter the trapezius muscle. Random fibers were evaluated histologically to determine fiber type (that is, motor vs sensory axons).
In addition to an innervation from the spinal accessory nerve, the authors also identified cervical nerve innervations of all trapezius muscles. For these innervations, 3 sides were found to have fibers derived from C-2 to C-4, 2 sides had fibers derived from C-2 to C-3, and 25 sides had fibers derived from C-3 to C-4. Fibers derived from C-2 to C-4 were classified as a Type I innervation, those from C-2 to C-3 were classified as a Type II innervation, and those from C-3 to C-4 were classified as a Type III innervation. Immunohistochemical analysis of fibers from each of these types confirmed the presence of motor axons.
Based on the authors' study, cervical nerves innervate the trapezius muscle with motor fibers. These findings support surgical and clinical experiences in which partial or complete trapezius function is maintained after injury to the spinal accessory nerve. The degree to which these nerves innervate this muscle, however, necessitates further study. Such information may be useful following nerve transfer procedures, denervation techniques for cervical dystonia, or sacrifice of the spinal accessory nerve due to pathological entities.