Metabolic-imaging of human glioblastoma live tumors: A new precision-medicine approach to predict tumor treatment response early.

Background: Glioblastoma (GB) is the most severe form of brain cancer, with a 12-15 month median survival. Surgical resection, temozolomide (TMZ) treatment, and radiotherapy remain the primary therapeutic options for GB, and no new therapies have been introduced in recent years. This therapeutic standstill is primarily due to preclinical approaches that do not fully respect the complexity of GB cell biology and fail to test efficiently anti-cancer treatments. Therefore, better treatment screening approaches are needed. In this study, we have developed a novel functional precision medicine approach to test the response to anticancer treatments in organoids derived from the resected tumors of glioblastoma patients. Methods: GB organoids were grown for a short period of time to prevent any genetic and morphological evolution and divergence from the tumor of origin. We chose metabolic imaging by NAD(P)H fluorescence lifetime imaging microscopy (FLIM) to predict early and non-invasively ex-vivo anti-cancer treatment responses of GB organoids. TMZ was used as the benchmark drug to validate the approach. Whole-transcriptome and whole-exome analyses were performed to characterize tumor cases stratification. Results: Our functional precision medicine approach was completed within one week after surgery and two groups of TMZ Responder and Non-Responder tumors were identified. FLIM-based metabolic tumor stratification was well reflected at the molecular level, confirming the validity of our approach, highlighting also new target genes associated with TMZ treatment and identifying a new 17-gene molecular signature associated with survival. The number of MGMT gene promoter methylated tumors was higher in the responsive group, as expected, however, some non-methylated tumor cases turned out to be nevertheless responsive to TMZ, suggesting that our procedure could be synergistic with the classical MGMT methylation biomarker. Conclusions: For the first time, FLIM-based metabolic imaging was used on live glioblastoma organoids. Unlike other approaches, ex-vivo patient-tailored drug response is performed at an early stage of tumor culturing with no animal involvement and with minimal tampering with the original tumor cytoarchitecture. This functional precision medicine approach can be exploited in a range of clinical and laboratory settings to improve the clinical management of GB patients and implemented on other cancers as well.

Single-Cell Molecular Characterization to Partition the Human Glioblastoma Tumor Microenvironment Genetic Background.

BACKGROUND: Glioblastoma (GB) is a devastating primary brain malignancy. The recurrence of GB is inevitable despite the standard treatment of surgery, chemotherapy, and radiation, and the median survival is limited to around 15 months. The barriers to treatment include the complex interactions among the different cellular components inhabiting the tumor microenvironment. The complex heterogeneous nature of GB cells is helped by the local inflammatory tumor microenvironment, which mostly induces tumor aggressiveness and drug resistance. METHODS: By using fluorescent multiple labeling and a DEPArray cell separator, we recovered several single cells or groups of single cells from populations of different origins from IDH-WT GB samples. From each GB sample, we collected astrocytes-like (GFAP+), microglia-like (IBA1+), stem-like cells (CD133+), and endothelial-like cells (CD105+) and performed Copy Number Aberration (CNA) analysis with a low sequencing depth. The same tumors were subjected to a bulk CNA analysis. RESULTS: The tumor partition in its single components allowed single-cell molecular subtyping which revealed new aspects of the GB altered genetic background. CONCLUSIONS: Nowadays, single-cell approaches are leading to a new understanding of GB physiology and disease. Moreover, single-cell CNAs resource will permit new insights into genome heterogeneity, mutational processes, and clonal evolution in malignant tissues.

Type II Fractures in Older Adults: Can They Be Treated Conservatively?: A Single-Center Experience and Review of the Literature.

BACKGROUND: Odontoid fractures are the most common acute cervical spinal fractures in the geriatric population. Their rate is increasing along with the rising age of the elderly population. Whereas conservative management with external immobilization is reported as the treatment of choice for type I and III odontoid fractures, there are no clear indications concerning the best treatment for type II fractures. In younger patients surgical management is considered the best choice, but in older adults the rate of good outcomes worsens and operative risk because of comorbidities increases. METHODS: We report our retrospective single-center experience with conservative treatment of type II odontoid fractures in an elderly population, focusing on both radiologic and functional outcomes to compare our results with the recent literature. RESULTS: Among the 21 selected subjects with a minimum follow-up of 18 months, 19 (90.5%) showed a satisfactory clinical outcome, with an adequate bony healing in 10 cases and nonsymptomatic pseudarthrosis in 9 patients. All these patients were satisfied with the conservative results and could stop use of the collar. Two patients (9.5%) did not show any improvement and had to keep the collar indefinitely. CONCLUSIONS: Our study was limited because it was a retrospective review, with a limited number of patients. Nevertheless, the clinical and radiologic outcomes of our patients differ from the results of other studies, suggesting that conservative management of these fractures in this population does not necessarily lead to a bad clinical outcome or delayed surgery.

A new quantitative method to assess disproportionately enlarged subarachnoid space (DESH) in patients with possible idiopathic normal pressure hydrocephalus: The SILVER index.

OBJECTIVES: Preoperative diagnosis of idiopathic normal-pressure hydrocephalus (iNPH) remains challenging. Recently, the presence of disproportionally enlarged subarachnoid spaces and hydrocephalus (DESH) on diagnostic images has been linked to clinical improvement after ventriculoperitoneal (VP) shunt placement. In this study we describe a new quantitative method to assess DESH on CT scans and to evaluate its prognostic value. PATIENTS AND METHODS: A multiplanar reconstruction software was used to retrospectively evaluate prospectively collected radiological data (CT scans) of 26 controls and 29 consecutive patients that underwent VP shunt placement for possible iNPH. The ratio between the areas of the sylvian fissure and the subarachnoid space at the vertex was calculated (SILVER index). The diagnostic accuracy of the SILVER index and the estimate of the best cut-point were assessed using ROC analysis. RESULTS: The mean value of the SILVER index was 11.52±14.27 in the study group and 1.68±0.98 in the control group (p-value<0.0001). The area under the ROC curve for the SILVER index was 0.903 (95% CI 0.813-0.994). A cut-off value for the SILVER index of 3.75 was extrapolated with a sensitivity and specificity of 0.828 and 0.962 respectively. CONCLUSIONS: The SILVER index is a reliable tool to easily quantify DESH on CT scans of patients with suspected iNPH. Its high sensitivity and specificity should encourage further investigations in order to confirm its clinical utility.

Use of near-infrared indocyanine videoangiography and Flow 800 in the resectioning of a spinal cord haemangioblastoma.

Haemangioblastomas are hypervascularized tumours. Their surgical management requires a complete resectioning and a prompt handling of the vascular inlets and outlets. The use of intraoperative indocyanine green video angiography (ICG-VAG) depicts the precise vascular pattern for the surgeon. Its use is safe and easy, and the procedure can be repeated during the operation. Here we present a case of spinal haemangioblastoma treated with the aid of intraoperative ICG-VAG and the Flow 800 software. The use of the Flow 800 allowed the surgeon to detect, at a glance, minimal changes in the vascular supply during the dissection. The colour-coded images generated by the Flow 800 increase the ICG-CAG sensitivity, improving the capability to detect changes in vascular patterns.