Expression of the progenitor marker NG2/CSPG4 predicts poor survival and resistance to ionising radiation in glioblastoma

Glioblastoma (GBM) is a highly aggressive brain tumour, where patients respond poorly to radiotherapy and exhibit dismal survival outcomes. The mechanisms of radioresistance are not completely understood. However, cancer cells with an immature stem-like phenotype are hypothesised to play a role in radioresistance. Since the progenitor marker neuron-glial-2 (NG2) has been shown to regulate several aspects of GBM progression in experimental systems, we hypothesised that its expression would influence the survival of GBM patients. Quantification of NG2 expression in 74 GBM biopsies from newly diagnosed and untreated patients revealed that 50% express high NG2 levels on tumour cells and associated vessels, being associated with significantly shorter survival. This effect was independent of age at diagnosis, treatment received and hypermethylation of the O6-methylguanine methyltransferase (MGMT) DNA repair gene promoter. NG2 was frequently co-expressed with nestin and vimentin but rarely with CD133 and the NG2 positive tumour cells harboured genetic aberrations typical for GBM. 2D proteomics of 11 randomly selected biopsies revealed upregulation of an antioxidant, peroxiredoxin-1 (PRDX-1), in the shortest surviving patients. Expression of PRDX-1 was associated with significantly reduced products of oxidative stress. Furthermore, NG2 expressing GBM cells showed resistance to ionising radiation (IR), rapidly recognised DNA damage and effectuated cell cycle checkpoint signalling. PRDX-1 knockdown transiently slowed tumour growth rates and sensitised them to IR in vivo. Our data establish NG2 as an important prognostic factor for GBM patient survival, by mediating resistance to radiotherapy through induction of ROS scavenging enzymes and preferential DNA damage signalling

Thymic stromal lymphopoietin does not activate human basophils.

International audienc

Longitudinal T1-weighted images and tumour growth.

<p>(<b>A</b>) Longitudinal axial post-contrast T1-weighted images of nude rats bearing U87MG tumours treated with combination NK+mAb9.2.27, NK cell monotherapy, mAb9.2.27 monotherapy, and vehicle controls, showing the same animal after 7 days, 17 days and 3 months post NK+mAb9.2.27 treatment. (<b>B</b>) Tumour volumes (#voxels) quantified on post-contrast T1-weighted images, before and after 7 and 17 days treatment. Data represents mean ±SEM of all animals treated.</p

Immunohistochemical staining and cell proliferation.

<p>(<b>A</b>, top panel) haematoxylin and eosin staining showing leucocyte packed necrosis in U87MG tumours treated with NK+mAb9.2.27 and mAb9.2.27 monotherapy, (arrows), Magnification 200X; Scale bar 100 µm. Cellular dense tumours treated with NK cell monotherapy and haemorrhaging control, untreated tumours. (<b>A</b>, middle panel) Ki67 staining of proliferating tumour cells (<b>A</b>, bottom panel). Magnification 200X; scale bar 100 µm. Tunel stained apoptotic cells, Magnification 200X; Scale bar 100 µm. (<b>B</b>) Quantified Ki67 labelling index, data represents mean ±SEM of all animals treated. (<b>C</b>) Quantified Tunel labelling index, data represents mean ±SEM of all animals treated,*p<0.05; **p<0.001, ***p<0.0001. D Ratio of proliferation: apoptosis index.</p

Expression of glial cell markers and NK cell ligands on U87MG tumour cells.

<p>(<b>A</b>) Mean Fluorescence Intensity (MFI) histograms and % cells expressing glial markers (GFAP,Nestin, Vimentin, and A2B5), <b>(B)</b> MFI histograms and % cells expressing class I HLA ligands (HLA-A,-B,-C), non-classical HLA-G and HLA-E, as well as HLA-DR,DP,DQ. <b>(C)</b> MFI histograms and % cells expressing ULBP 1, 3, 2/5/6, MICA and MICB activating ligands. Dark histograms represent negative control, light histograms represent stained cells.</p

Perfusion parameters and maps.

<p>(<b>A</b>) Elevated extravascular extracellular volume fraction v_e, in NK+mAb9.2.27 compared to monotherapy animals from cohort 1 (received 1 million NK cells), *p<0.05, **p<0.001 ***p<0.0001. (<b>A left panels</b>) Parametric maps visualising intratumoral heterogeneity in v_e in representative control, monotherapy and NK+mAb9.2.27 treated animals at 7 days. Intensity scale shows minimum (blue voxels) and maximum (red voxels) intensity levels. (<b>B</b>) Increased interstitial extracellular volume fraction, v_e, elevated in NK+mAb9.2.27 compared to NK cells and mAb9.2.27 monotherapy animals, and decreased v_e in NK cell monotherapy compared to untreated control from cohort 2 (treated with 2 million NK cells), *p<0.05, **p<0.001, ***p<0.0001. (<b>B, left panels</b>) Parametric maps visualising intratumoural heterogeneity in v_e in control, monotherapy and NK+mAb9.2.27 treated animals at 17 days. (<b>C</b>) Significant association between ADC and v_e in the NK+mAb9.2.27 (R² = 0.798, p = 0.041) and NK cell monotherapy animals (R² = 0.993, p = 0.004). Graphs in A and B represent estimated marginal mean±95% confidence intervals.</p

Longitudinal T2-weighted images.

<p>Longitudinal corresponding axial T2-weighted images of same animals as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108414#pone-0108414-g002" target="_blank">Figure 2</a>. Nude rats bearing U87MG tumours treated with combination NK+mAb9.2.27, NK cell monotherapy, mAb9.2.27 monotherapy, and vehicle controls, showing the same animal after 7 days, 17 days and 3 months post NK+mAb9.2.27 treatment.</p

AllergoOncology: Microbiota in allergy and cancer-A European Academy for Allergy and Clinical Immunology position paper

The microbiota can play important roles in the development of human immunity and the establishment of immune homeostasis. Lifestyle factors including diet, hygiene, and exposure to viruses or bacteria, and medical interventions with antibiotics or anti-ulcer medications, regulate phylogenetic variability and the quality of cross talk between innate and adaptive immune cells via mucosal and skin epithelia. More recently, microbiota and their composition have been linked to protective effects for health. Imbalance, however, has been linked to immune-related diseases such as allergy and cancer, characterized by impaired, or exaggerated immune tolerance, respectively. In this AllergoOncology position paper, we focus on the increasing evidence defining the microbiota composition as a key determinant of immunity and immune tolerance, linked to the risk for the development of allergic and malignant diseases. We discuss novel insights into the role of microbiota in disease and patient responses to treatments in cancer and in allergy. These may highlight opportunities to improve patient outcomes with medical interventions supported through a restored microbiome

Inborn errors of OAS–RNase L in SARS-CoV-2–related multisystem inflammatory syndrome in children

International audienceMultisystem inflammatory syndrome in children (MIS-C) is a rare and severe condition that follows benign COVID-19. We report autosomal recessive deficiencies of OAS1 , OAS2 , or RNASEL in five unrelated children with MIS-C. The cytosolic double-stranded RNA (dsRNA)–sensing OAS1 and OAS2 generate 2′-5′-linked oligoadenylates (2-5A) that activate the single-stranded RNA–degrading ribonuclease L (RNase L). Monocytic cell lines and primary myeloid cells with OAS1, OAS2, or RNase L deficiencies produce excessive amounts of inflammatory cytokines upon dsRNA or severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) stimulation. Exogenous 2-5A suppresses cytokine production in OAS1-deficient but not RNase L–deficient cells. Cytokine production in RNase L–deficient cells is impaired by MDA5 or RIG-I deficiency and abolished by mitochondrial antiviral-signaling protein (MAVS) deficiency. Recessive OAS–RNase L deficiencies in these patients unleash the production of SARS-CoV-2–triggered, MAVS-mediated inflammatory cytokines by mononuclear phagocytes, thereby underlying MIS-C