Background supratotal resection is advocated in lower-grade-gliomas (LGGs) based on theoretical advantages, but with limited verification of functional risk and data on oncological outcomes. We assessed the association of supratotal resection in molecular-defined LGGs with oncological outcomes. Methods 460 presumptive LGGs included; 404 resected; 347 were LGGs, 319 IDH-mutated, 28 wildtype. All patients had clinical, imaging, molecular data. Resection aimed at supratotal resection without any patient or tumor a-priori selection. The association of Extent-of-Resection (EOR), categorized on volumetric-FLAIR-images as residual-tumor-volume, along with post-surgical-management with Progression-free-survival (PFS), malignant-progression-free-survival (MPFS), and Overall-Survival (OS) assessed by univariate, multivariate, propensity-score-analysis. The study mainly focused on IDH-mutated-LGGs, the “typical LGGs”. Results Median follow-up:6.8 years(IQR:5-8). Out of 319 IDH-mutated-LGGs, 190 (59.6%) progressed, median PFS:4.7 years(95%CI:4–5.3). Total and supratotal resection obtained in 39% and 35% of patients of IDH1-mutated tumors. In IDH-mutated, most patients in partial/subtotal group progressed, 82.4% in total, only 6 (5.4%) in supratotal. Median PFS was 29 months(95%CI:25-36) in subtotal, 46 months(95%CI:38-48) in total, while at 92 months, PFS in supratotal was 94.0%. There was no association with molecular-subtypes and grade. At random-forest-analysis, PFS strongly associated with EOR,RT, previous treatment. In the propensity-score analysis, EOR associated with PFS (HR,0.03;95%CI,0.01-0.13). MPFS occurred in 32.1% of subtotal-total groups; 1 event in supratotal. EOR, grade-III, previous treatment correlated to MPFS. At random-forest analysis, OS associated with EOR as well. Conclusions Supratotal resection strongly associated with PFS, MPFS and OS in LGGs, regardless of molecular subtypes and grade, right from the beginning of clinical presentation.

MGMT (O⁶-methylguanine-DNA methyltransferase) promoter hypermethylation is a helpful prognostic marker for chemotherapy of gliomas, although with some controversy for low-grade tumors. The objective of this study was to retrospectively investigate MGMT promoter hypermethylation status for a series of 350 human brain tumors, including 275 gliomas of different malignancy grade, 21 glioblastoma multiforme (GBM) cell lines, and 75 non-glial tumors. The analysis was performed by methylation-specific PCR and capillary electrophoresis. MGMT expression at the protein level was also evaluated by both immunohistochemistry (IHC) and western blotting analysis. Associations of MGMT hypermethylation with IDH1/IDH2 mutations, EGFR amplification, TP53 mutations, and 1p/19q co-deletion, and the prognostic significance of these, were investigated for the gliomas. MGMT promoter hypermethylation was identified in 37.8% of gliomas, but was not present in non-glial tumors, with the exception of one primitive neuroectodermal tumor (PNET). The frequency was similar for all the astrocytic gliomas, with no correlation with histological grade. Significantly higher values were obtained for oligodendrogliomas. MGMT promoter hypermethylation was significantly associated with IDH1/IDH2 mutations (P = 0.0207) in grade II–III tumors, whereas it had a borderline association with 1p deletion (P = 0.0538) in oligodendrogliomas. No other association was found. Significant correlation of MGMT hypermethylation with MGMT protein expression was identified by IHC in GBMs and oligodendrogliomas (P = 0.0001), but not by western blotting. A positive correlation between MGMT protein expression, as detected by either IHC or western blotting, was also observed. The latter was consistent with MGMT promoter hypermethylation status in GBM cell lines. In low-grade gliomas, MGMT hypermethylation, but not MGMT protein expression, was associated with a trend, only, toward better survival, in contrast with GBMs, for which it had favorable prognostic significance.

It is already well known that hypermethylation of the O6-methylguanine DNA methyltransferase (MGMT) gene promoter is a predictive biomarker of response to temozolomide treatment and of favorable outcomes in terms of overall survival (OS) and progression-free survival (PFS) in glioblastoma (GBM) patients. Nevertheless, MGMT methylation status has not currently been introduced into routine clinical practice, as the choice of the ideal technique and tissue sample specimen is still controversial. The aim of this study was to compare 2 analytical methods, methylation-specific polymerase chain reaction (MSP) and pyrosequencing (PSQ), and their use on 2 different tissue type samples, snap-frozen and formalin-fixed paraffin-embedded (FFPE), obtained from a single-center and uniformly treated cohort of 46 GBM patients. We obtained methylation data from all frozen tissues, while no results were obtained for 5 FFPE samples. The highest concordance for methylation was found on frozen tissues (88.5%, 23/26 samples), using PSQ (76.7%, 23/30 samples). Moreover, we confirmed that OS and PFS for patients carrying methylation of the MGMT promoter were longer than for patients with an unmethylated promoter. In conclusion, we considered MSP a limited technique for FFPE tissues due to the high risk of false-positive results; in contrast, our data indicated PSQ as the most powerful method to stratify methylated/unmethylated patients as it allows reaching quantitative results with high sensitivity and specificity. Furthermore, frozen tumor tissues were shown to be the best specimens for MGMT methylation analysis, due to the low DNA degradation and homogeneity in methylation throughout the tumor.

Abstract:Patients with Alzheimer's disease (AD) are characterized by an altered sensitivity to cortisol-mediated modulation of circulating lymphocytes. Longitudinal studies are needed to address the clinical applicability of these abnormalities as prognostic factors. Therefore, we designed a longitudinal study to address the clinical applicability of physiologic modulation of Natural Killer (NK) cell activity as a prognostic factor in AD. NK activity was assessed as baseline measurement and in response to modulation by cortisol at 10 -6 M. To verify the immunophysiological integrity of the NK cell population, we tested augmentation of NK cytotoxicity by human recombinant interleukin (IL)-2 (100 IU/ml) as control. The response to modulation by cortisol or by IL-2 was significantly greater in patients with AD. Based on change in the Mini-Mental State score at entry and at 18 months, patients with AD could be assigned to a "fast progression" (Δ > 2 points) or to a "slow progression" group (Δ < 2 points). The change in the response of NK cytotoxic activity to cortisol, and the strength of the association of this parameter with circulating activated T cells in time was greater in patients with Fast Progression vs. Slow Progression AD. These results suggest that changes in the response of NK cells to negative (e.g., cortisol) or positive modifiers (e.g., IL-2) follow progression of AD.

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