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Molecular glioblastomas (i.e. without the histological but with the molecular characteristics of IDH-wild-type glioblastoma) frequently lack contrast enhancement, which can wrongly lead to suspect a lower-grade glioma. Herein, we aimed to assess the diagnostic value of gyriform infiltration as an imaging marker for molecular glioblastomas.Two independent investigators reviewed the MRI scans from patients with newly diagnosed gliomas for the presence of a gyriform infiltration defined as an elective cortical hypersignal on MRI FLAIR sequence. Diagnostic test performance of this sign for the diagnosis of molecular glioblastoma were calculated.A total of 426 patients were included, corresponding to 31 molecular glioblastoma, 294 IDH-wild-type glioblastoma, 50 IDH-mutant astrocytoma, and 51 IDH-mutant 1p19q-codeleted oligodendroglioma. A gyriform infiltration was observed in 16/31 (52%) molecular glioblastoma, 40/294 (14%) IDH-wild-type glioblastoma, and none of the IDH-mutant glioma. All the 56 gyriform-infiltration-positive tumors were IDH-wild-type and all but two had a TERT promoter mutation. The inter-rater agreement was good (κ = 0.69, p 0.001). The sensitivity, specificity, positive predictive value and negative predictive value of the presence of a gyriform infiltration for the diagnosis of molecular glioblastoma were 52%, 90%, 29%, 96%, respectively. The median overall survival was better for gyriform-infiltration-negative patients compared to gyriform-infiltration-positive patients in the whole series and in patients with non-enhancing lesions (n = 95) (25.6 vs 16.9 months, p = 0.005 and 20.2 months vs not reached, p 0.001).Gyriform infiltration is a specific imaging marker of molecular glioblastomas that can help distinguishing these tumors from IDH-mutant lower-grade gliomas.

International audience; Adult tumors diagnosed as cerebellar glioblastoma (cGBM) are rare and their optimal classification remains to be determined. The aim of this study was to identify subgroups of cGBM based on targeted molecular analysis. cGBM diagnosed between 2003 and 2017 were identified from the French Brain Tumor Database and reviewed according to the WHO 2021 classification. The following molecular alterations were studied: IDH1/2, H3F3A, FGFR1, BRAF, TERT promoter mutations, EGFR amplification, MGMT promoter methylation, and alternative lengthening of telomere status. DNA methylation profile was assessed in a subset of cases. Eighty-three cGBM were included and could be classified into 6 mutually exclusive subgroups associated with median age at diagnosis (MA) and prognosis: TERT-mutant and/or EGFR-amplified tumors (n=22, 26.5%, MA=62 y, median overall survival [OS]=4 mo), H3K27M-mutant tumors (n=15, 18.1%, MA=48 y, median OS=8 mo), mitogen-activated protein kinases (MAPK) pathway-activated tumors (FGFR1, BRAF mutation, or occurring in neurofibromatosis type I patients, n=15, 18.1%, MA=48 y, median OS=57 mo), radiation-associated tumors (n=5, 6%, MA=47 y, median OS=5 mo), IDH-mutant tumors (n=1), and unclassified tumors (n=25, 30.1%, MA=63 y, median OS=17 mo). Most MAPK pathway-activated tumors corresponded to high-grade astrocytomas with piloid features based on DNA methylation profiling. In multivariate analysis, MAPK pathway-activating alterations, ATRX loss of expression, and alternative lengthening of telomere positivity were independently associated with a better outcome and TERT/EGFR alterations with a worse outcome. cGBM display an important intertumoral heterogeneity. Targeted molecular analysis enables to classify the majority of tumors diagnosed as cGBM into mutually exclusive and clinically relevant subgroups. The presence of MAPK pathway alterations is associated with a much better prognosis.

Pleomorphic xanthoastrocytoma (PXA) is classified as an astrocytic glioma occurring most often in children or young adults. Molecular alterations in PXA are not fully known, especially those associated with tumor progression. We describe a patient with several relapses of a PXA. The tumor showed an acquired ATRX loss through tumor evolution. We tested alternative lengthening of telomeres (ALT) with the C-circle test. While the test was negative in the first tumor, a high circle activity was detected in the last relapse, suggesting an acquired ALT phenotype. Our data not only confirm previous findings of the possible occurrence of ATRX mutations in PXA but also suggest that this alteration is linked to PXA progression. In small biopsies, tumors with ATRX loss, without IDH or histone mutation, pathologists should consider the diagnosis of PXA, especially if associated with BRAF V600E mutation, CDKN2A deletion, and ALT.

BACKGROUND Biopsies in patients with a suspected glioma are occasionally nondiagnostic. OBJECTIVE To explore the utility of molecular testing in this setting by determining whether IDH1 and TERT promoter (pTERT) mutations could be detected in nondiagnostic biopsies from glioma patients. METHODS Using SNaPshot polymerase chain reaction, we retrospectively assessed IDH1 and pTERT mutation status in nondiagnostic biopsies from 28 glioma patients. RESULTS The nondiagnostic biopsy (needle biopsy n = 25, open or endoscopic biopsy n = 3) consisted of slight glial cell hypercellularity, hemorrhage, and/or necrosis. After another biopsy (n = 23) or a subsequent surgical resection (n = 5) the diagnosis was an IDH1-wildtype (WT) pTERT-mutant glioma (glioblastoma n = 16, astrocytoma n = 4), an IDH1-mutant pTERT-mutant oligodendroglioma (n = 1), an IDH1-mutant pTERT-WT astrocytoma (n = 1), and an IDH1-WT pTERT-WT glioblastoma (n = 6). An IDH1 mutation was identified in the nondiagnostic biopsies of the 2 IDH-mutant gliomas, and a pTERT mutation in the nondiagnostic biopsies of 16 out of the 21 of pTERT mutant-gliomas (76%). Overall, an IDH1 and/or a pTERT mutation were detected in 17 out of 28 (61%) of nondiagnostic biopsies. Retrospective analysis of the nondiagnostic biopsies based on these results and on imaging characteristics suggested that a new biopsy could have been avoided in 6 patients in whom a diagnosis of "molecular glioblastoma" could have been done with a high level of confidence. CONCLUSION In the present series, IDH1 and pTERT mutations could be detected in a high proportion of nondiagnostic biopsies from glioma patients. Molecular testing may facilitate the interpretation of nondiagnostic biopsies in patients with a suspected glioma.

Background In glioma, TERT promoter mutation and loss of ATRX (ATRX loss) are associated with reactivation of telomerase or alternative lengthening of telomeres (ALT), respectively, i.e. the two telomere maintenance mechanisms (TMM). Strangely, 25% of gliomas have been reported to display neither or both of these alterations. Materials and methods The C-circle (CC) assay was adapted to tumor (formalin-fixed paraffin-embedded and frozen) and blood samples to investigate the TMM. Results We constructed a CC-based algorithm able to identify the TMM and reported a sensitivity of 100% and a specificity of 97.3% (n = 284 gliomas). By combining the TMM, the mutational status of the isocitrate dehydrogenase 1/2 (IDH) gene (IDHmt), and the histological grading, we propose a new classification tool: TeloDIAG. This classification defined five subtypes: tOD, tLGA, tGBM_IDHmt, tGBM, and tAIV, corresponding to oligodendroglioma, IDHmt low-grade astrocytoma, IDHmt glioblastoma, and IDHwt glioblastoma (GBM), respectively; the last class gathers ALT+ IDHwt gliomas that tend to be related to longer survival (21.2 months) than tGBM (16.5 months). The TeloDIAG was 99% concordant with the World Health Organization classification (n = 312), and further modified the classification of 55 of 144 (38%) gliomas with atypical molecular characteristics. As an example, 14 of 69 (20%) of TERTwt, ATRXwt, and IDHwt GBM were actually tAIV. Outstandingly, CC in blood sampled from IDHmt astrocytoma patients was detected with a sensitivity of 56% and a specificity of 97% (n = 206 gliomas and 30 healthy donors). Conclusion The TeloDIAG is a new, simple, and effective tool helping in glioma diagnosis and a promising option for liquid biopsy.

BACKGROUND Diffuse astrocytic gliomas, IDH wildtype, with molecular features of glioblastoma (molecular glioblastomas) are associated with a poor prognosis. We previously found that these tumors frequently display gyriform infiltration, defined as areas of elective cortical hypersignal on MRI FLAIR sequence. The objective of the present study was to assess the diagnostic value of gyriform infiltration as an imaging marker for these tumors. MATERIAL AND METHODS MRI scans from 430 patients with newly diagnosed glioma (molecular glioblastoma n = 31, IDH wildtype glioblastoma n = 298, IDH-mutant astrocytoma n = 50, IDH-mutant and 1p19q codeleted oligodendroglioma n= 51) were evaluated for the presence of a gyriform infiltration by 2 independent reviewers. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated to assess the performance of the presence of a gyriform infiltration for identifying molecular glioblastoma. RESULTS A gyriform infiltration was observed in 16/31 (52%) patients with a molecular glioblastoma, 40/298 (13%) patients with an IDH-wildtype glioblastoma but in none of the patients with an IDH-mutant astrocytomas or an IDH-mutant and 1p19q codeleted oligodendroglioma. Among the 56 patients with a gyriform infiltration, 54 patients had an IDH wildtype pTERT mutant glioma and 2 an IDH wildtype pTERT wildtype glioma. Interrater agreement was good (κ= 0.68, P < 0.001). Specificity, sensitivity, PPV and NPV of the presence of a gyriform infiltration for the diagnosis of molecular glioblastoma were 90%, 29%, 52% and 96% and for the diagnosis of an IDHwt pTERT mutant glioma were 97%, 15%, 96% and 20%. The presence of a gyriform infiltration was associated with a worse prognosis in the entire cohort (13.6 months vs 29.3 months, P = .001). CONCLUSION Gyriform infiltration is a specific imaging marker of molecular glioblastomas and IDH wildtype pTERT mutant diffuse gliomas.

International audience; The Bcl-xL apoptosis inhibitor plays a major role in vertebrate development. In addition to its effect on apoptosis, Bcl-xL is also involved in cell migration and mitochondrial metabolism. These effects may favour the onset and dissemination of metastasis. However, the underlying molecular mechanisms remain to be fully understood. Here we focus on the control of cell migration by Bcl-xL in the context of breast cancer cells. We show that Bcl-xL silencing led to migration defects in Hs578T and MDA-MB231 cells. These defects were rescued by re-expressing mitochondria-addressed, but not endoplasmic reticulum-addressed, Bcl-xL. The use of BH3 mimetics, such as ABT-737 and WEHI-539 confirmed that the effect of Bcl-xL on migration did not depend on interactions with BH3-containing death accelerators such as Bax or BH3-only proteins. In contrast, the use of a BH4 peptide that disrupts the Bcl-xL/VDAC1 complex supports that Bcl-xL by acting on VDAC1 permeability contributes to cell migration through the promotion of reactive oxygen species production by the electron transport chain. Collectively our data highlight the key role of Bcl-xL at the interface between cell metabolism, cell death, and cell migration, thus exposing the VDAC1/Bcl-xL interaction as a promising target for anti-tumour therapy in the context of metastatic breast cancer.

Adult IDH-wildtype astrocytomas with TERT promoter mutations (TERTp) are associated with a poor prognosis.To analyze the radiological presentation and natural history of adult IDH-wildtype astrocytomas with TERTp.We retrospectively reviewed the characteristics of 40 IDH-wildtype TERTp-mutant astrocytomas (grade II n = 19, grade III n = 21) and compared them to those of 114 IDH-mutant lower grade gliomas (LGG), of 92 IDH-wildtype TERTp-mutant glioblastomas, and of 15 IDH-wildtype TERTp-wildtype astrocytomas.Most cases of IDH-wildtype TERTp-mutant astrocytomas occurred in patients aged50 yr (88%) and presented as infiltrative lesions without contrast enhancement (73%) that were localized in the temporal and/or insular lobes (37.5%) or corresponded to a gliomatosis cerebri (43%). Thalamic involvement (33%) and extension to the brainstem (27%) were frequently observed, as was gyriform infiltration (33%). This radiological presentation was different from that of IDH-mutant LGG, IDH-wildtype TERTp-mutant glioblastomas, and IDH-wildtype TERTp-wildtype astrocytomas. Tumor evolution before treatment initiation was assessable in 17 cases. Ten cases demonstrated a rapid growth characterized by the apparition of a ring-like contrast enhancement and/or a median velocity of diametric expansion (VDE) ≥8 mm/yr but 7 cases displayed a slow growth (VDE8 mm/yr) that could last several years before anaplastic transformation. Median overall survival of IDH-wildtype TERTp-mutant astrocytomas was 27 mo.IDH-wildtype TERTp-mutant astrocytomas typically present as nonenhancing temporo-insular infiltrative lesions or as gliomatosis cerebri in patients aged50 yr. In the absence of treatment, although rapid tumor growth is frequent, an initial falsely reassuring, slow growth can be observed.

BACKGROUND Adult cerebellar glioblastomas (cGBM) are very rare and recent studies have shown that they constitute a heterogeneous group of gliomas. The aim of the present study was to characterize the prevalence and prognostic significance of major driver molecular alterations in a large series of cGBM. MATERIAL AND METHODS Adults with histologically proven cGBM diagnosed between 2003 and 2017 were identified from the French brain tumor database and the Club de Neuro-Oncologie of the Société Française de Neurochirurgie. Tumors were reviewed and reclassified according to WHO 2016. Targeted sequencing was performed, including determination of H3F3A, TERTp, IDH1/2, FGFR1, BRAF and EGFR status. RESULTS A total of 83 adult patients (median age 57 years) with cGBM (hemispheric n= 47, vermian n=14 or both n=22) were identified. Median overall survival was 10 months. Main molecular alterations observed were TERT promoter, H3F3A K27M, hotspot FGFR1 (N546 and K656), BRAF mutations, EGFR amplification and ATRX loss of expression in 19.2%, 18.8%, 10.9%, 2.6%, 19.5% and 22.7% of patients, respectively. cGBM could be classified into 6 mutually exclusive subgroups associated with age at diagnosis and prognosis: pTERT and/or EGFR amplified tumors (n=22, 26.5%, median age = 62 years, median OS = 4 months), H3K27M-mutant tumors (n=15, 18.1%, median age = 48 years, median OS =8 months), tumors with MAPK pathway activating mutations (FGFR1, BRAF) or occurring in NF1 patients (n=15, 18.1%, median age = 41 years, median OS = 57 months), radiation-induced tumors (n=5, 6%, median age = 47 years, median OS = 5 months), IDH-mutant tumors (n=1) and unclassified tumors (n=25, 30.1%, median age = 63 years, median OS = 17 months). In multivariate analysis, MAPK activating mutations and ATRX loss of expression were independently associated with a better outcome and pTERT/EGFR alterations with a worse outcome. CONCLUSION About 18% of tumors diagnosed as cGBM harbor actionable MAPK activating genetic alterations. Targeted sequencing enables to classify these tumors into clinically relevant subgroups.

All cancer cells need to maintain functional telomeres to sustain continuous cell division and proliferation. In human diffuse gliomas, functional telomeres are maintained due either to reactivation of telomerase expression, the main pathway in most cancer types, or to activation of a mechanism called the alternative lengthening of telomeres (ALT). The presence of IDH1/2 mutations (IDH-mutant) together with loss of ATRX expression (ATRX-lost) are frequently associated with ALT in diffuse gliomas. However, detection of ALT, anda fortioriits quantification, are rarely, if ever, measured in neuropathology laboratories. We measured the level of ALT activity using the previously described quantitative “C-circle” assay and analyzed it in a well characterized cohort of 104 IDH-mutant and ATRX-lost adult diffuse gliomas. We report that in IDH-mutant ATRX-lost anaplastic astrocytomas, the intensity of ALT was inversely correlated with age (p

BACKGROUND We previously showed that TERT promotor (pTERT) mutations can be detected in a high proportion of non-diagnostic biopsies from glioma patients using SNaPshot PCR. The aim of the present study was to assess the diagnostic value of digital droplet PCR (ddPCR) in this setting. MATERIAL AND METHODS pTERT mutations were retrospectively assessed using SNaPshot PCR and ddPCR in the non-diagnostic biopsies of 14 adult patients who underwent an initial non-diagnostic needle biopsy that led to perform another biopsy that finally demonstrated a pTERT-mutant glioma. RESULTS Median age at diagnosis was 69 years and the median interval between the initial and final biopsy was 22 days. The initial biopsy consisted of unspecific infiltrating glial cells (n=10), hemorrhage, necrosis and/or fibrosis (n=4). Using SNaPshot PCR, the final biopsy found an IDH-WT pTERT-mutant glioma in 12 cases (glioblastoma n=12), an IDH-mutant pTERT-mutant oligodendroglioma in one case and an IDH-WT pTERT-WT glioblastoma in one case. Retrospectively, a pTERT mutation was identified by both ddPCR and SNaPshot in the non-diagnostic biopsies in 85% (11 /13) of pTERT mutant-gliomas. No mutation was detected in the non-diagnostic biopsies of IDH-WT pTERT-WT gliomas using either technique. Overall, a pTERT mutation were detected in 79% (11/14) of non-diagnostic biopsies. CONCLUSION Digital Droplet PCR, as SNaPshot PCR, enables the identification of pTERT mutations in a high rate of non-diagnostic biopsies from glioma patients. The sensitivity of both techniques seems similar in this setting.

BACKGROUND H3 G34-mutant high-grade gliomas occur primarily in children but can also be encountered in adults. The aim of the present study was to describe the characteristics of H3 G34-mutant high-grade gliomas in adults. MATERIAL AND METHODS We analysed the characteristics of 17 adult H3 G34-mutant high-grade gliomas and compared them with those of adult grade IV gliomas with H3 K27M mutation (n=21), IDH mutation (n=42) and TERT promoter (pTERT) mutation (n=88). RESULTS Median age at diagnosis in H3 G34-mutant gliomas was 25 years (range: 19–33 y). All tumors had a hemispheric location. The radiological presentation was suggestive of a high-grade glioma in 6 patients but in 10 patients it initially suggested another diagnosis due to absent or faint contrast enhancement (n=9) or the presence of an initial intratumoral haemorrhage (n=1). In non- or faint contrast-enhancing cases, diffusion-weighted imaging (DWI) was more helpful to suspect an aggressive tumor than MR spectroscopy and perfusion MRI. All H3 G34-mutant gliomas were classified as IDH wildtype glioblastomas. PNET-like foci were observed in 8 cases and most cases were immunonegative for ATRX (92%) and Olig2 (81%) and demonstrated MGMT promoter methylation (82%). The clinical, radiological and histological presentation of H3 G34-mutant gliomas was different from that of midline H3 K27M-mutant diffuse gliomas and from that of IDH-mutant and IDH-wildtype pTERT-mutant glioblastomas. Median overall survival of G34-mutant gliomas was 23.9 months compared to 19.6 months (p=0.16), 13.7 months (p=0.45) and 58 months (p=2.3x10-10) in H3 K27M-mutant diffuse gliomas, IDHwt pTERT-mutant and IDH-mutant glioblastomas, respectively. CONCLUSION Adults H3 G34-mutant high-grade gliomas are associated with distinct characteristics and a poor prognosis. DWI seems helpful to identify cases that present as non-contrast enhancing lesions and can wrongly suggest another diagnosis in these young patients.

Genomic duplications and fusion involving BRAF and KIAA1549 that create fusion proteins with constitutive B-RAF kinase activity are a hallmark of pilocytic astrocytomas (PAs). The detection of KIAA1549–BRAF fusion transcripts is of paramount importance to classify these tumors and to identify patients who could benefit from BRAF inhibitors. In a clinical setting, the available material for molecular analysis from these pediatric tumors is often limited to formalin-fixed paraffin-embedded (FFPE) tissue. The aim of the present study was to develop a new method to detect the three most frequent KIAA1549–BRAF fusion transcripts, 15–9, 16–11, and 16–9, where numbers refer to the exons fused together, using a FFPE-compatible multiplex quantitative reverse transcription polymerase chain reaction (qRT-PCR). We compared performance of the assay to a reference singleplex method on a collection of 46 FFPE PAs. The results showed that both methods are comparable. The multiplex method had an overall 97% sensitivity and 100% specificity compared to the singleplex method, and agreement between the two techniques was almost perfect (Cohen’s kappa: 0.97). There was no evidence of a significant difference between the qRT-PCR efficiencies of the multiplex technique and of the singleplex assay for all fusion transcripts and for GAPDH, the latter used as a reference gene. The multiplex method consumed four times less complementary DNA (cDNA), cost less, and required half the hands-on technical time. The results show that it could be beneficial to implement the multiplex method in a clinical setting, where samples presenting low quantity of degraded RNA are not unusual.

The Bcl-xL apoptosis inhibitor plays a major role in vertebrate development. In addition to its effect on apoptosis, Bcl-xL is also involved in cell migration and mitochondrial metabolism. These effects may favour the onset and dissemination of metastasis. However, the underlying molecular mechanisms remain to be fully understood. Here we focus on the control of cell migration by Bcl-xL in the context of breast cancer cells. We show that Bcl-xL silencing led to migration defects in Hs578T and MDA-MB231 cells. These defects were rescued by re-expressing mitochondria-addressed, but not endoplasmic reticulum-addressed, Bcl-xL. The use of BH3 mimetics, such as ABT-737 and WEHI-539 confirmed that the effect of Bcl-xL on migration did not depend on interactions with BH3-containing death accelerators such as Bax or BH3-only proteins. In contrast, the use of a BH4 peptide that disrupts the Bcl-xL/VDAC1 complex supports that Bcl-xL by acting on VDAC1 permeability contributes to cell migration through the promotion of reactive oxygen species production by the electron transport chain. Collectively our data highlight the key role of Bcl-xL at the interface between cell metabolism, cell death, and cell migration, thus exposing the VDAC1/Bcl-xL interaction as a promising target for anti-tumour therapy in the context of metastatic breast cancer.

BACKGROUND: Non-diagnostic biopsies are a recurrent issue in patients with a suspected brain tumor. Herein, in order to explore the utility of molecular testing in this setting, we determined whether IDH and TERT promoter (pTERT) mutations can be detected in non-diagnostic biopsies from glioma patients. MATERIAL AND METHODS: Using Snapshot PCR, we retrospectively assessed IDH and pTERT mutation status in 28 adult glioma patients in whom a first non-diagnostic biopsy had led to perform a second biopsy. RESULTS: Median age at diagnosis was 65 years and median delay between the first and second biopsy 21 days. The first biopsy consisted of non characterizable infiltrated glial cells (n=19), hemorrhage (n=4), necrosis (n=2) or normal tissue (n=3). The second biopsy demonstrated an IDH-wildtype glioblastoma (n=22), an IDH-wildtype astrocytoma (n=4), an IDH-mutant oligodendroglioma (n=1) and an IDH-mutant astrocytoma (n=1). A pTERT mutation was present in 21 cases. Retrospectively, the same IDH and pTERT mutations were identified in the non-diagnostic biopsies of the 2 patients with an IDH-mutant glioma and of 12 out of 21 patients with a pTERT-mutant glioma (57%). Overall an IDH and/or a pTERT mutation were detected in the non-diagnostic biopsies of 13 out of the 22 IDH-mutant and/or pTERT-mutant gliomas (59%) and in 13 out of the 28 cases included in the present series (46%). CONCLUSION: IDH and pTERT mutations can be detected in a high percentage of non-diagnostic biopsies from glioma patients supporting a role for molecular testing in the interpretation of non-diagnostic biopsies from patients with a suspected brain tumor

BACKGROUND: Adult IDH-wildtype astrocytomas with TERT promoter mutations (TERTp) are associated with a poor prognosis. The aim of the present study was to analyze their radiological presentation and natural history. MATERIAL AND METHODS: We retrospectively reviewed the characteristics of 40 IDH-wildtype TERTp-mutant astrocytomas (grade II n=19, grade III n=21) and compared them to those of 114 IDH-mutant lower grade gliomas (LGG), of 92 IDH-wildtype TERTp-mutant glioblastomas and of 15 IDH-wildtype TERTp-wildtype astrocytomas. RESULTS: Most cases of IDH-wildtype TERTp-mutant astrocytomas occurred in patients aged >50 years (88%) and presented as infiltrative lesions without contrast enhancement (73%) that were localized in the temporal and/or insular lobes (37.5%) or corresponded to a gliomatosis cerebri (43%). Thalamic involvement (33%) and extension to the brainstem (27%) were frequently observed, as was gyriform infiltration (33%). This radiological presentation was different from that of IDH-mutant LGG, IDH-wildtype TERTp-mutant glioblastomas, and IDH-wildtype TERTp-wildtype astrocytomas. Tumor evolution before treatment initiation was assessable in 17 cases. Ten cases demonstrated a rapid growth characterized by the apparition of a ring-like contrast enhancement and/or a median velocity of diametric expansion (VDE) ≥ 8 mm/year but 7 cases displayed a slow growth (VDE < 8 mm/year) that could last several years before anaplastic transformation. Median overall survival of IDH-wildtype TERTp-mutant astrocytomas was 27 months. CONCLUSION: IDH-wildtype TERTp-mutant astrocytomas typically present as non-enhancing temporo-insular infiltrative lesions or as gliomatosis cerebri in patients aged >50 years. In the absence of treatment, although rapid tumor growth is frequent, an initial falsely reassuring, slow growth can be observed.

Adult cerebellar glioblastomas (cGBM) are rare and their characteristics remain to be fully described. We analyzed the characteristics of 17 adult patients with cGBM and compared them to a series of 103 patients presenting a supra-tentorial glioblastoma (stGBM). The mean age at GBMc diagnosis was 53.4 years (range 28–77). A history of neurofibromatosis type I was noted in 3 patients. cGBM were hemispheric in 10 patients (58.8%), only vermian in 4 patients (23.5%), and both vermian and hemispheric in 3 patients (17.7%). A H3 K27M mutation was identified in 3/14 patients, a TERT promoter mutation in 3/14 patients and a methylated MGMT promoter in 3/14 patients. None of the patients (0/14) harbored an EGFR amplification, an IDH or a BRAF mutation. Association with neurofibromatosis type I and H3K27M mutations were mutually exclusive. Compared with stGBM, cGBM occurred in younger patients (53.4 vs. 63.2, p = 0.02), were more frequently associated with neurofibromatosis type I (18 vs. 1%, p = 0.009) and with a H3 K27M mutation (21 vs. 3%, p = 0.02). They also tended to have a more frequent multifocal presentation at diagnosis (21 vs. 4.3%, p = 0.06), more frequently resulted in leptomeningeal or intra-axial metastasis (44.5 vs. 5%, p = 0.002) and were associated with a shorter median overall survival (5.9 vs. 14.2 months, p = 0.004). The present study suggests that adult cGBM differ from their supra-tentorial counterpart and constitute a heterogeneous group of IDH wild-type gliomas with at least two subgroups, one associated with H3K27M mutations and the other with neurofibromatosis type I.

Nowadays, head and neck squamous cell carcinoma (HNSCC) treatment failure is mostly explained by locoregional progression or intrinsic radioresistance. Radiotherapy (RT) has recently evolved with the emergence of heavy ion radiations or new fractionation schemes of photon therapy, which modify the dose rate of treatment delivery. The aim of the present study was then to evaluate the in vitro influence of a dose rate variation during conventional RT or carbon ion hadrontherapy treatment in order to improve the therapeutic care of patient. In this regard, two HNSCC cell lines were irradiated with photons or 72 MeV/n carbon ions at a dose rate of 0.5, 2, or 10 Gy/min. For both radiosensitive and radioresistant cells, the change in dose rate significantly affected cell survival in response to photon exposure. This variation of radiosensitivity was associated with the number of initial and residual DNA double-strand breaks (DSBs). By contrast, the dose rate change did not affect neither cell survival nor the residual DNA DSBs after carbon ion irradiation. As a result, the relative biological efficiency at 10% survival increased when the dose rate decreased. In conclusion, in the RT treatment of HNSCC, it is advised to remain very careful when modifying the classical schemes toward altered fractionation. At the opposite, as the dose rate does not seem to have any effects after carbon ion exposure, there is less need to adapt hadrontherapy treatment planning during active system irradiation.

International audience; Cancer cells protect their telomere ends from erosion through reactivation of telomerase or by using the Alternative Lengthening of Telomere (ALT) mechanism that depends on homologous recombination. Chronic lymphocytic leukaemia (CLL) B cells are characterized by almost no telomerase activity, shelterin deregulation and telomere fusions. To characterize telomeric maintenance mechanisms in B-CLL patients, we measured their telomere length, telomerase expression and the main hallmarks of the ALT activity i.e. C-circle concentration, an extra-chromosomal telomere repeat (ECTR), and the level of telomeric sister chromatid exchange (T-SCE) rate. Patients showed relative homogenous telomere length although almost no TERT transcript and nearly no C-circle were evidenced. Nevertheless, compared with normal B cells, B-CLL cells showed an increase in T-SCE rate that was correlated with a strong down-regulation of the topoisomerase III alpha (TOP3A) expression, involved in the dissolution of Holliday Junctions (HJ), together with an increased expression of SLX1A, SLX4, MUS81 and GEN1, involved in the resolution of HJ. Altogether, our results suggest that the telomere maintenance mechanism of B-CLL cells do not preferentially use telomerase or ALT. Rather, the rupture of the dissolvasome/resolvasome balance may increase telomere shuffling that could homogenize telomere length, slowing telomere erosion in this disease

At the time of diagnosis, 60% of patients with head and neck squamous cell carcinoma (HNSCC) present tumors in an advanced stage (III-IV) of disease and 80% will relapse within the first two years post-treatment, due to their frequent radio(chemo)resistance. To identify new molecular targets and companion biomarkers, we have investigated the miRNome of 75 stage III-IV oropharynx tumors without relapse (R) or with loco-regional relapse (non-responder, NR) within two years post-treatment. Interestingly, miR-422a was significantly downregulated in NR tumors, in agreement with the increase in cell proliferation and adhesion induced by miR-422a inhibition in vitro. Furthermore, we identified CD73/NT5E oncogene as target of miR-422a. Indeed, modulation of the endogenous level of miR-422a inversely influences the expression and the enzymatic activity of CD73. Moreover, knocking down CD73 mimics the effects of miR-422a upregulation. Importantly, in tumors, miR-422a and CD73 expression levels are inversely correlated, and both are predictive of relapse free survival - especially considering loco(regional) recurrence - in vitro two independent cohorts of advanced oropharynx or HNSCC (N=255) tumors. In all, we reported, for the first time, that MiR-422a and its target CD73 are involved in early loco(regional) recurrence of HNSCC tumors and are new targets for personalized medicine.

International audience; The higher biological effect of Carbon ions hadrontherapy (C+) is explained by the nature of the DNA damages. It is known that cell response to γ-irradiation (γ-IR), but not to C+, is correlated with telomere length in different type of cancer cells. Here, we propose that this " telomeric effect " must result from an effect of ROS in γ-IR compared to C+.

BACKGROUND: Non-diagnostic biopsies are a recurrent issue in patients with a suspected brain tumor. Herein, in order to explore the utility of molecular testing in this setting, we determined whether IDH and TERT promoter (pTERT) mutations can be detected in non-diagnostic biopsies from glioma patients. METHODS: Using Snapshot PCR, we retrospectively assessed IDH and pTERT mutation status in 28 adult glioma patients in whom a first non-diagnostic biopsy had led to perform a second biopsy. RESULTS: Median age at diagnosis was 65 years and median delay between the first and second biopsy 21 days. The first biopsy consisted of not characterizable infiltrated glial cells (n=19), hemorrhage (n=4), necrosis (n=2) or normal tissue (n=3). The second biopsy demonstrated an IDH-wildtype glioblastoma (n=22), an IDH-wildtype astrocytoma (n=4), an IDH-mutant oligodendroglioma (n=1) and an IDH-mutant astrocytoma (n=1). A pTERT mutation was present in 21 cases. Retrospectively, the same IDH and pTERT mutations were identified in the non-diagnostic biopsies of the 2 patients with an IDH-mutant glioma and of 12 out of 21 patients with a pTERT-mutant glioma (57%). Overall an IDH and/or a pTERT mutation were detected in the non-diagnostic biopsies of 13 out of the 22 IDH-mutant and/or pTERT-mutant gliomas (59%) and in 13 out of the 28 cases included in the present series (46%). CONCLUSION: IDH and pTERT mutations can be detected in a high percentage of non-diagnostic biopsies from glioma patients supporting a role for molecular testing in the interpretation of non-diagnostic biopsies from patients with a suspected brain tumor.

International audience; The multiform glioblastoma (GBM) is a heterogeneous and highly invasive entity, making it the most aggressive brain tumor. The standard-of-care for glioblastoma consists of surgical resection, radiotherapy and chemotherapy, but despite the recent improvement of therapeutic protocols, the recurrence seems inevitable. Due to a better dose localization in the tumor volume and a greater Relative Biological Efficiency (RBE), carbon ion therapy seems to be a promising alternative to conventional radiotherapy. However, to optimize individual treatment by hadrontherapy the exact carbon equivalent dose needs to be determined from data obtained after photon irradiation. Therefore, molecular and cellular investigations of GBM are required in order to improve the prediction and treatment of brain tumors. In this study, we have examined the radiobiological features of 11 human glioma cell lines displaying gradual radiosensitivity, following photon- or carbon-therapy in order to optimize and secure antiglioma strategies. Independent of p53 or O6-methylguanine-DNA methyltransferase (MGMT) status, all cell lines respond to both types of radiation by a G2/M phase arrest followed by the appearance of mitotic catastrophe, which is concluded by a ceramide-dependent-apoptotic cell death. Statistical analyses demonstrate that: (i) the surviving fraction at 2Gy (SF2) and the dose for 10% survival (D10) photon values are correlated with that obtained in response to carbon ions; (ii) regardless of p53, MGMT status, and radiosensitivity, the release of ceramide is associated with the induction of late apoptosis; (iii) the appearance of polyploid cells after photon irradiation could predict the RBE to carbon ions.The present study clearly provides a consistent database of the cellular and molecular response of glioblastoma cell lines to photon irradiation, and to the best of our knowledge represents the largest archive for carbon ion hadrontherapy response of glioblastoma cells. This type of archive is intended to customize the treatment of patients by allowing the development of new predictive mathematical models for the response of tumors to radiation, and ultimately could improve hadrontherapy treatment plans.

Background Glioblastoma (GBM) is the most frequent and aggressive type of adult brain tumor. Most GBMs express telomerase; a high level of intra-tumoral telomerase activity (TA) is predictive of poor prognosis. Thus, telomerase inhibitors are promising options to treat GBM. These inhibitors increase the response to radiotherapy (RT), in vitro as well as in vivo. Since typical treatments for GBM include RT, our objective was to evaluate the efficiency of Imetelstat (TA inhibitor) combined with RT. Findings We used a murine orthotopic model of human GBM (N = 8 to11 mice per group) and μMRI imaging to evaluate the efficacy of Imetelstat (delivered by intra-peritoneal injection) alone and combined with RT. Using a clinically established protocol, we demonstrated that Imetelstat significantly: (i) inhibited the TA in the very center of the tumor, (ii) reduced tumor volume as a proportion of TA inhibition, and (iii) increased the response to RT, in terms of tumor volume regression and survival increase. Conclusions Imetelstat is currently evaluated in refractory brain tumors in young patients (without RT). Our results support its clinical evaluation combined with RT to treat GBM. Electronic supplementary material The online version of this article (doi:10.1186/s12943-015-0376-3) contains supplementary material, which is available to authorized users.

La télomérase, composée de différentes sous unité catalytique (TERT), ribonucléique (TERC) et protéique (Dyskérine). Il s’agit d’une enzyme vitale pour maintien de la taille des extrémités des chromosomes ou télomères. En effet, elle permet la synthèse de nouvelles répétitions télomériques lors du processus de réplication de l’ADN. Cette capacité d’allongement des télomères est détournée dans des phénomènes pathologiques comme le cancer et en physiologie, une taille trop faible (limite d’Hayflick) de télomères entraîne l’arrêt du cycle cellulaire c’est ce que l’on appelle la sénescence réplicative. Pour échapper à ce processus, les cellules tumorales ré-exprime sa sous unité catalytique TERT et acquièrent ainsi une immortalité réplicative. Il est important de noter que physiologiquement un raccourcissement des télomères est associé à un vieillissement cellulaire. Dans cette thèse, nous explorons aussi les autres fonctions biologiques de la télomères et démontrons les raisons pour lesquelles le domaine de la thérapeutique est en expansion concernant les thérapies antitélomérase. Nous recenserons les différentes stratégies ciblant cette enzyme avec les dérivés naturels, les inhibiteurs pharmacologiques de la télomérase (les oligonucléotides anti-sens), mais aussi les ligands des Gquadruplexe composants les télomères ou encore l’essors de thérapie immunologique anti-TERT. Ainsi nous mettons en lumière le fort intérêt de cette enzyme tant pour la prise en charge des patients atteints de cancer mais aussi ceux atteints de maladies métaboliques que l’on peut mettre en lien avec le processus de vieillissement ellulaire