Subventricular zone involvement is associated with worse outcome in glioma WHO grade 2 depending on molecular markers

Neural stem cells within the subventricular zone were identified as cells of origin driving growth of high-grade gliomas, and anatomical involvement of the subventricular zone has been associated with an inferior clinical outcome. Whether the association between poor outcome and subventricular zone involvement also applies to glioma of lower grades is unclear. We therefore analysed a retrospective cohort of 182 patients with glioma grade 2 (according to the WHO 2016 classification) including 78 individuals (43%) with subventricular zone involvement. Patients with and without subventricular zone involvement did not differ in regard to demographics, histopathology, and molecular markers. Notably, subventricular zone involvement was a negative prognostic marker for malignant progression and overall survival on uni- and multivariate analysis. When patients were stratified according to the cIMPACT-NOW update 6, subventricular zone involvement was negatively associated with outcome in IDH-wildtype astrocytomas and 1p19q-codeleted oligodendrogliomas but not in IDH-mutant astrocytomas. Collectively, subventricular zone involvement may represent a risk factor for worse outcome in glioma WHO grade 2 depending on the molecular tumor signature. The present data confirm the relevance of molecular glioma classifications as proposed by the cIMPACT-NOW update 6. These findings warrant evaluation in prospective cohorts

Bright polar mesospheric clouds formed by main engine exhaust from the space shuttle's final launch

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/94747/1/jgrd18009.pd

Deciphering sources of PET signals in the tumor microenvironment of glioblastoma at cellular resolution

Various cellular sources hamper interpretation of positron emission tomography (PET) biomarkers in the tumor microenvironment (TME). We developed an approach of immunomagnetic cell sorting after in vivo radiotracer injection (scRadiotracing) with three-dimensional (3D) histology to dissect the cellular allocation of PET signals in the TME. In mice with implanted glioblastoma, translocator protein (TSPO) radiotracer uptake per tumor cell was higher compared to tumor-associated microglia/macrophages (TAMs), validated by protein levels. Translation of in vitro scRadiotracing to patients with glioma immediately after tumor resection confirmed higher single-cell TSPO tracer uptake of tumor cells compared to immune cells. Across species, cellular radiotracer uptake explained the heterogeneity of individual TSPO-PET signals. In consideration of cellular tracer uptake and cell type abundance, tumor cells were the main contributor to TSPO enrichment in glioblastoma;however, proteomics identified potential PET targets highly specific for TAMs. Combining cellular tracer uptake measures with 3D histology facilitates precise allocation of PET signals and serves to validate emerging novel TAM-specific radioligands

DNA methylation profiling to predict recurrence risk in meningioma: development and validation of a nomogram to optimize clinical management

Abstract Background Variability in standard-of-care classifications precludes accurate predictions of early tumor recurrence for individual patients with meningioma, limiting the appropriate selection of patients who would benefit from adjuvant radiotherapy to delay recurrence. We aimed to develop an individualized prediction model of early recurrence risk combining clinical and molecular factors in meningioma. Methods DNA methylation profiles of clinically annotated tumor samples across multiple institutions were used to develop a methylome model of 5-year recurrence-free survival (RFS). Subsequently, a 5-year meningioma recurrence score was generated using a nomogram that integrated the methylome model with established prognostic clinical factors. Performance of both models was evaluated and compared with standard-of-care models using multiple independent cohorts. Results The methylome-based predictor of 5-year RFS performed favorably compared with a grade-based predictor when tested using the 3 validation cohorts (ΔAUC = 0.10, 95% CI: 0.03–0.018) and was independently associated with RFS after adjusting for histopathologic grade, extent of resection, and burden of copy number alterations (hazard ratio 3.6, 95% CI: 1.8–7.2, P &lt; 0.001). A nomogram combining the methylome predictor with clinical factors demonstrated greater discrimination than a nomogram using clinical factors alone in 2 independent validation cohorts (ΔAUC = 0.25, 95% CI: 0.22–0.27) and resulted in 2 groups with distinct recurrence patterns (hazard ratio 7.7, 95% CI: 5.3–11.1, P &lt; 0.001) with clinical implications. Conclusions The models developed and validated in this study provide important prognostic information not captured by previously established clinical and molecular factors which could be used to individualize decisions regarding postoperative therapeutic interventions, in particular whether to treat patients with adjuvant radiotherapy versus observation alone. </jats:sec

Beta2-adrenoceptor stimulation suppresses TLR9-dependent IFNA1 secretion in human peripheral blood mononuclear cells.

INTRODUCTION: IFNA1 (interferon alpha) is a key cytokine regulating the activity of numerous immune cells. Plasmacytoid dendritic cells (pDCs) as natural interferon-producing cells play critical roles as sensors of pathogens and link innate to adaptive immunity. CpG motifs within DNA sequences activating toll-like receptor 9 (TLR9) are the main stimuli eliciting IFNA1 secretion from pDCs. Adrenergic substances are capable of differentially modulating the response from various immune cells. Hence, the aim of this study was to examine how adrenoceptor stimulation influences TLR9-induced IFNA1 secretion from human pDCs. METHODS: PBMCs generated from human whole blood and pDCs enriched from buffy coats were stimulated with LPS and CpG-ODN 2336 in the presence or absence of epinephrine and different adrenoceptor antagonists. Secretion of TNF and IFNA1 was measured by ELISA. Flow cytometry was used to determine efficacy of pDC enrichment and adrenoceptor expression of PBMC subsets. The influence of modified IFNA1 secretion on NK cell activity was evaluated using a colorimetric tumor cell lysis assay. RESULTS: TLR9-induced IFNA1 secretion as well as TLR4-induced TNF secretion from PBMCs was dose-dependently attenuated by coincubation with epinephrine. Combination with different specific adrenoceptor antagonists revealed that this effect was mediated by the adrenoceptor β2 (ADRB2). Since flow cytometric analysis could exclude the presence of ADRB2 on pDCs, highly enriched pDCs lacked any visible impact of adrenoceptor stimulation on TLR9-induced IFNA1 release. Combination of pDCs with PBMCs restored the effect, even when they were separated by a permeable membrane. Suppression of TLR9-mediated IFNA1 secretion from PBMCs by adrenoceptor stimulation reduced the lytic activity of NK cells on K562 tumor cells. CONCLUSION: We provide insights into the underlying mechanisms of the interrelation between immune responses and pharmacological agents widely used in clinical practice. Our results have implications for the future treatment of human patients, in which the endogenous immune response plays a pivotal role, such as during viral infections, inflammatory diseases and cancers

Expression of ADRB2 in human PBMCs, enrichement of pDCs.

<p>(A, B) PBMCs were generated from freshly-drawn blood from healthy human donors. After staining with antibodies against CD123, CD304, CD14 and ADRB2 or an isotype control, cells were incubated with FITC-labeled secondary anti-rabbit-IgG antibody, and ADRB2 expression was assessed by flow cytometry. (A) Histograms showing the fluorescence signal for ADRB2-FITC in PBMCs and PBMC subsets. B) Percentage of ADRB2⁺ cells within pDC- or monocyte-subpopulations within PBMC. (<b>C</b>, <b>D</b>) PBMCs were isolated from freshly prepared buffy coats from healthy human donors and pDCs were subsequently enriched by MACS technique. (<b>C</b>) Cells were labeled with anti-human-CD303 antibody, and the frequency of CD303⁺ cells was determined by flow cytometry. Dot plots show percentage of CD303⁺ cells before (upper panel) and after MACS procedure (lower panel) (gated on live cells). Right: Histogram showing the fluorescence signal for CD303 before and after MACS procedure. (<b>D</b>) Following MACS procedure, enriched pDCs and the pDC-depleted PBMCs were stimulated with PBS (vehicle) or CpG ODN 2336 (2.5 or 5.0 µg/ml). After 24 hours, IFNA1 release into the supernatant was measured by ELISA; p<0.005 for enriched pDCs vs. pDC-depleted PBMCs. CpG = CpG ODN 2336; SSC = side scatter. *** p<0.005.</p

Interaction of enriched pDCs and PBMCs.

<p>PBMCs were generated from freshly-drawn blood from healthy human donors. PDCs were enriched by MACS technique from PBMCs derived from freshly prepared buffy coats from healthy human donors. (A) Enriched pDCs (white bars) or pDCs supplemented with PBMCs (pDC∶PBMC ratio 1∶10, grey bars) were stimulated with PBS (vehicle), CpG ODN 2336 (1.25 µg/ml) or CpG ODN in the presence of epinephrine (10⁻⁶ mol/l). After 24 hours, IFNA1 release into the supernatant was measured by ELISA. Data is presented as percentage of CpG ODN-induced IFNA1 secretion. (B) In the same experiment, enriched pDCs (white bars) were compared to enriched pDCs seeded into the upper compartment of a two-chamber transwell system containing PBMCs in the lower compartment (pDC∶PBMC ratio 1∶10, hatched bars). Cells were stimulated with PBS (vehicle), CpG ODN 2336 (1.25 µg/ml) or CpG ODN in the presence of epinephrine (10⁻⁶ mol/l). After 24 hours, IFNA1 release into the supernatant was measured by ELISA. Data is presented as percentage of CpG ODN-induced IFNA1 secretion. (C) Absolute IFNA1 levels of corresponding conditions in (A). CpG = CpG ODN 2336; epi = epinephrine. * p<0.05; *** p<0.005; statistical comparisons are indicated by brackets.</p