bims-malgli 2021-04-18 papers

bims-malgli

Biomed News

on Biology of malignant gliomas

Issue of 2021–04–18
twenty-one papers selected by
Oltea Sampetrean, Keio University

TNFα secreted by glioma associated macrophages promotes endothelial activation and resistance against anti-angiogenic therapy.
The white matter is a pro-differentiative niche for glioblastoma.
Sex Differences in Glioblastoma Immunotherapy Response.
Kinomic profile in patient-derived glioma cells during hypoxia reveals c-MET-PI3K dependency for adaptation.
Multiscale modeling of glioma pseudopalisades: contributions from the tumor microenvironment.
Meclofenamate causes loss of cellular tethering and decoupling of functional networks in glioblastoma.
Cannabidiol converts NFκB into a tumor suppressor in glioblastoma with defined antioxidative properties.
De novo purine biosynthesis is a major driver of chemoresistance in glioblastoma.
Elevated Asparagine Biosynthesis Drives Brain Tumor Stem Cell Metabolic Plasticity and Resistance to Oxidative Stress.
Tumor immune landscape of paediatric high-grade gliomas.
PI3Kγ inhibition suppresses microglia/TAM accumulation in glioblastoma microenvironment to promote exceptional temozolomide response.
Brd4-bound enhancers drive cell-intrinsic sex differences in glioblastoma.
Novel targetable FGFR2 and FGFR3 alterations in glioblastoma associate with aggressive phenotype and distinct gene expression programs.
Modified RANO (mRANO), iRANO, and standard RANO response to convection-enhanced delivery of IL4R-targeted immunotoxin MDNA55 in recurrent glioblastoma.
Dual targeting of the epigenome via FACT complex and histone deacetylase is a potent treatment strategy for DIPG.
Metabolic imaging detects elevated glucose flux through the pentose phosphate pathway associated with TERT expression in low-grade gliomas.
Turning strength into weakness: protein degradation and autophagy as therapeutic targets in Glioblastoma?
Global phosphoproteomics reveals DYRK1A regulates CDK1 activity in glioblastoma cells.
Glioblastomas acquire myeloid-affiliated transcriptional programs via epigenetic immunoediting to elicit immune evasion.
Long-term survivors of glioblastoma are a unique group of patients lacking universal characteristic features.
Association of preoperative seizures with tumor metabolites quantified by magnetic resonance spectroscopy in gliomas.

Acta Neuropathol Commun. 2021 Apr 14. 9(1): 67

TNFα secreted by glioma associated macrophages promotes endothelial activation and resistance against anti-angiogenic therapy.

Qingxia Wei, Olivia Singh, Can Ekinci, Jaspreet Gill, Mira Li, Yasin Mamatjan, Shirin Karimi, Severa Bunda, Sheila Mansouri, Kenneth Aldape, Gelareh Zadeh.

  One of the most prominent features of glioblastoma (GBM) is hyper-vascularization. Bone marrow-derived macrophages are actively recruited to the tumor and referred to as glioma-associated macrophages (GAMs) which are thought to provide a critical role in tumor neo-vascularization. However, the mechanisms by which GAMs regulate endothelial cells (ECs) in the process of tumor vascularization and response to anti-angiogenic therapy (AATx) is not well-understood. Here we show that GBM cells secrete IL-8 and CCL2 which stimulate GAMs to produce TNFα. Subsequently, TNFα induces a distinct gene expression signature of activated ECs including VCAM-1, ICAM-1, CXCL5, and CXCL10. Inhibition of TNFα blocks GAM-induced EC activation both in vitro and in vivo and improve survival in mouse glioma models. Importantly we show that high TNFα expression predicts worse response to Bevacizumab in GBM patients. We further demonstrated in mouse model that treatment with B20.4.1.1, the mouse analog of Bevacizumab, increased macrophage recruitment to the tumor area and correlated with upregulated TNFα expression in GAMs and increased EC activation, which may be responsible for the failure of AATx in GBMs. These results suggest TNFα is a novel therapeutic that may reverse resistance to AATx. Future clinical studies should be aimed at inhibiting TNFα as a concurrent therapy in GBMs.

Keywords:  Anti-angiogenic therapy; Endothelial cells activation; Glioblastoma; Glioma-associated macrophages; TNFα

DOI:  https://doi.org/10.1186/s40478-021-01163-0
Nat Commun. 2021 04 12. 12(1): 2184

The white matter is a pro-differentiative niche for glioblastoma.

Lucy J Brooks, Melanie P Clements, Jemima J Burden, Daniela Kocher, Luca Richards, Sara Castro Devesa, Leila Zakka, Megan Woodberry, Michael Ellis, Zane Jaunmuktane, Sebastian Brandner, Gillian Morrison, Steven M Pollard, Peter B Dirks, Samuel Marguerat, Simona Parrinello.

Glioblastomas are hierarchically organised tumours driven by glioma stem cells that retain partial differentiation potential. Glioma stem cells are maintained in specialised microenvironments, but whether, or how, they undergo lineage progression outside of these niches remains unclear. Here we identify the white matter as a differentiative niche for glioblastomas with oligodendrocyte lineage competency. Tumour cells in contact with white matter acquire pre-oligodendrocyte fate, resulting in decreased proliferation and invasion. Differentiation is a response to white matter injury, which is caused by tumour infiltration itself in a tumoursuppressive feedback loop. Mechanistically, tumour cell differentiation is driven by selective white matter upregulation of SOX10, a master regulator of normal oligodendrogenesis. SOX10 overexpression or treatment with myelination-promoting agents that upregulate endogenous SOX10, mimic this response, leading to niche-independent pre-oligodendrocyte differentiation and tumour suppression in vivo. Thus, glioblastoma recapitulates an injury response and exploiting this latent programme may offer treatment opportunities for a subset of patients.

DOI: https://doi.org/10.1038/s41467-021-22225-w
Neuromolecular Med. 2021 Apr 17.

Sex Differences in Glioblastoma Immunotherapy Response.

Juyeun Lee, Kristen Kay, Katie Troike, Manmeet S Ahluwalia, Justin D Lathia.

  Glioblastoma (GBM), the most common primary malignant brain tumor, remains difficult to treat and shares phenotypes, including an aberrant immune response, with other neurological disorders. Understanding the cellular and molecular mechanisms underlying this pathological immune response remains a priority, particularly as standard of care for advanced cancers evolves to include immunotherapies, which have yet to show strong clinical efficacy in GBM. Epidemiological evidence supports a sex difference in GBM, with increased prevalence in males, and recent studies identified differences between males and females ranging from genetic aberrations to cellular programs. Sex differences have also been identified in immune response, and in this mini-review, we present these differences to highlight potential sex-specific cellular and molecular mechanisms that underly GBM growth and response to immunotherapies. These sex differences offer an opportunity to understand GBM pathogenesis and extend beyond GBM to other tumors and neurological disorders to inform the development of next-generation therapies.

Keywords:  Glioma; Immune system; Immunotherapy; Sex differences

DOI:  https://doi.org/10.1007/s12017-021-08659-x
Theranostics. 2021 ;11(11): 5127-5142

Kinomic profile in patient-derived glioma cells during hypoxia reveals c-MET-PI3K dependency for adaptation.

Hong Sheng Cheng, Charlie Marvalim, Pengcheng Zhu, Cheng Lui Daniel Law, Zhi Yan Jeremy Low, Yuk Kien Chong, Beng Ti Ang, Carol Tang, Nguan Soon Tan.

  Hypoxic microenvironment is a hallmark of solid tumors, especially glioblastoma. The strong reliance of glioma-propagating cells (GPCs) on hypoxia-induced survival advantages is potentially exploitable for drug development. Methods: To identify key signaling pathways for hypoxia adaptation by patient-derived GPCs, we performed a kinase inhibitor profiling by screening 188 small molecule inhibitors against 130 different kinases in normoxia and hypoxia. Potential kinase candidates were prioritized for in vitro and in vivo investigations using a ranking algorithm that integrated information from the kinome connectivity network and estimated patients' survival based on expression status. Results: Hypoxic drug screen highlighted extensive modifications of kinomic landscape and a crucial functionality of c-MET-PI3K. c-MET inhibitors diminished phosphorylation of c-MET and PI3K in GPCs subjected to hypoxia, suggesting its role in the hypoxic adaptation of GPCs. Mechanistically, the inhibition of c-MET and PI3K impaired antioxidant defense, leading to oxidative catastrophe and apoptosis. Repurposed c-MET inhibitors PF04217903 and tivantinib exhibited hypoxic-dependent drug synergism with temozolomide, resulting in reduced tumor load and growth of GPC xenografts. Detailed analysis of bulk and single-cell glioblastoma transcriptomes associates the cellular subpopulation over-expressing c-MET with inflamed, hypoxic, metastatic, and stem-like phenotypes. Conclusions: Thus, our "bench to bedside (the use of patient-derived GPCs and xenografts for basic research) and back (validation with independent glioblastoma transcriptome databases)" analysis unravels the novel therapeutic indications of c-MET and PI3K/Akt inhibitors for the treatment of glioblastoma, and potentially other cancers, in the hypoxic tumor microenvironment.

Keywords:  HIF-1α; PI3K; glioblastoma; oxidative stress; tumor microenvironment

DOI:  https://doi.org/10.7150/thno.54741
J Math Biol. 2021 Apr 12. 82(6): 49

Multiscale modeling of glioma pseudopalisades: contributions from the tumor microenvironment.

Pawan Kumar, Jing Li, Christina Surulescu.

  Gliomas are primary brain tumors with a high invasive potential and infiltrative spread. Among them, glioblastoma multiforme (GBM) exhibits microvascular hyperplasia and pronounced necrosis triggered by hypoxia. Histological samples showing garland-like hypercellular structures (so-called pseudopalisades) centered around the occlusion site of a capillary are typical for GBM and hint on poor prognosis of patient survival. We propose a multiscale modeling approach in the kinetic theory of active particles framework and deduce by an upscaling process a reaction-diffusion model with repellent pH-taxis. We prove existence of a unique global bounded classical solution for a version of the obtained macroscopic system and investigate the asymptotic behavior of the solution. Moreover, we study two different types of scaling and compare the behavior of the obtained macroscopic PDEs by way of simulations. These show that patterns (not necessarily of Turing type), including pseudopalisades, can be formed for some parameter ranges, in accordance with the tumor grade. This is true when the PDEs are obtained via parabolic scaling (undirected tissue), while no such patterns are observed for the PDEs arising by a hyperbolic limit (directed tissue). This suggests that brain tissue might be undirected - at least as far as glioma migration is concerned. We also investigate two different ways of including cell level descriptions of response to hypoxia and the way they are related .

Keywords:  Directed/undirected tissue; Glioblastoma; Global existence; Hypoxia-induced tumor behavior; Kinetic transport equations; Long time behavior; Multiscale modeling; Pseudopalisade patterns; Reaction-diffusion-taxis equations; Uniqueness; Upscaling

DOI:  https://doi.org/10.1007/s00285-021-01599-x
Neuro Oncol. 2021 Apr 17. pii: noab092. [Epub ahead of print]

Meclofenamate causes loss of cellular tethering and decoupling of functional networks in glioblastoma.

Matthias Schneider, Lea Vollmer, Anna-Laura Potthoff, Vidhya M Ravi, Bernd O Evert, Mohummad A Rahman, Shahin Sarowar, Jan Kueckelhaus, Paulina Will, David Zurhorst, Kevin Joseph, Julian P Maier, Nicolas Neidert, Paolo d'Errico, Melanie Meyer-Luehmann, Ulrich G Hofmann, Andreas Dolf, Paolo Salomoni, Erdem Güresir, Per Ø Enger, Martha Chekenya, Torsten Pietsch, Patrick Schuss, Oliver Schnell, Mike-Andrew Westhoff, Jürgen Beck, Hartmut Vatter, Andreas Waha, Ulrich Herrlinger, Dieter H Heiland.

   BACKGROUND: Glioblastoma cells assemble to a syncytial communicating network based on tumor microtubes (TMs) as ultra-long membrane protrusions. The relationship between network architecture and transcriptional profile remains poorly investigated. Drugs that interfere within this syncytial connectivity such as meclofenamate (MFA) may be highly attractive for glioblastoma therapy.
METHODS: In a human neocortical slice model using glioblastoma cell populations of different transcriptional signatures, three-dimensional tumor networks were reconstructed and TM-based intercellular connectivity was mapped on the base of two-photon imaging data. MFA was used to modulate morphological and functional connectivity; downstream effects of MFA treatment were investigated by RNA sequencing and fluorescence-activated cell sorting (FACS) analysis.
RESULTS: TM-based network morphology strongly differed between the transcriptional cellular subtypes of glioblastoma and was dependent on axon guidance molecule expression. MFA revealed both a functional and morphological demolishment of glioblastoma network architectures which was reflected by a reduction of TM-mediated intercellular cytosolic traffic as well as a breakdown of TM length. RNA sequencing confirmed a downregulation of NCAM and axon guidance molecule signaling upon MFA treatment. Loss of glioblastoma communicating networks was accompanied by a failure in the upregulation of genes that are required for DNA repair in response to TMZ-treatment and culminated in profound treatment response to TMZ-mediated toxicity.
CONCLUSION: The capacity of TM formation reflects transcriptional cellular heterogeneity. MFA effectively demolishes functional and morphological TM-based syncytial network architectures. These findings might pave the way to a clinical implementation of MFA as a TM-targeted therapeutic approach.

Keywords:  Glioblastoma; intercellular network architecture; meclofenamate; tumor microtubes

DOI:  https://doi.org/10.1093/neuonc/noab092
Neuro Oncol. 2021 Apr 17. pii: noab095. [Epub ahead of print]

Cannabidiol converts NFκB into a tumor suppressor in glioblastoma with defined antioxidative properties.

Marie N M Volmar, Jiying Cheng, Haitham Alenezi, Sven Richter, Alisha Haug, Zonera Hassan, Maria Goldberg, Yuping Li, Mengzhuo Hou, Christel Herold-Mende, Cecile L Maire, Katrin Lamszus, Charlotte Flüh, Janka Held-Feindt, Gaetano Gargiulo, Geoffrey J Topping, Franz Schilling, Dieter Saur, Günter Schneider, Michael Synowitz, Joel A Schick, Roland E Kälin, Rainer Glass.

   BACKGROUND: The transcription factor NFκB drives neoplastic progression of many cancers including primary brain tumors (glioblastoma; GBM). Precise therapeutic modulation of NFκB activity can suppress central oncogenic signalling pathways in GBM, but clinically applicable compounds to achieve this goal have remained elusive.
METHODS: In a pharmacogenomics study with a panel of transgenic glioma cells we observed that NFκB can be converted into a tumor suppressor by the non-psychotropic cannabinoid Cannabidiol (CBD). Subsequently, we investigated the anti-tumor effects of CBD, which is used as an anticonvulsive drug (Epidiolex) in pediatric neurology, in a larger set of human primary GBM stem-like cells (hGSC). For this study we performed pharmacological assays, gene expression profiling, biochemical and cell-biological experiments. We validated our findings using orthotopic in vivo models and bioinformatics analysis of human GBM-datasets.
RESULTS: We found that CBD promotes DNA binding of the NFκB subunit RELA and simultaneously prevents RELA-phosphorylation on serine-311, a key residue which permits genetic transactivation. Strikingly, sustained DNA binding by RELA lacking phospho-serine 311 was found to mediate hGSC cytotoxicity. Widespread sensitivity to CBD was observed in a cohort of hGSC defined by low levels of reactive oxygen-species (ROS), while high ROS-content in other tumors blocked CBD induced hGSC death. Consequently, ROS levels served as predictive biomarker for CBD-sensitive tumors.
CONCLUSIONS: This evidence demonstrates how a clinically approved drug can convert NFκB into a tumor suppressor and suggests a promising repurposing option for GBM-therapy.

Keywords:  GBM therapy; NFB (Nuclear factor kappa-light-chain-enhancer of activated B cells); RELA (v-rel avian reticuloendotheliosis viral oncogene homolog A; also designated p65 or NFKB3); preclinical study; stem-like GBM cells

DOI:  https://doi.org/10.1093/neuonc/noab095
Brain. 2021 Apr 14. pii: awab020. [Epub ahead of print]

De novo purine biosynthesis is a major driver of chemoresistance in glioblastoma.

Jack M Shireman, Fatemeh Atashi, Gina Lee, Eunus S Ali, Miranda R Saathoff, Cheol H Park, Sol Savchuk, Shivani Baisiwala, Jason Miska, Maciej S Lesniak, C David James, Roger Stupp, Priya Kumthekar, Craig M Horbinski, Issam Ben-Sahra, Atique U Ahmed.

  Glioblastoma is a primary brain cancer with a near 100% recurrence rate. Upon recurrence, the tumour is resistant to all conventional therapies, and because of this, 5-year survival is dismal. One of the major drivers of this high recurrence rate is the ability of glioblastoma cells to adapt to complex changes within the tumour microenvironment. To elucidate this adaptation's molecular mechanisms, specifically during temozolomide chemotherapy, we used chromatin immunoprecipitation followed by sequencing and gene expression analysis. We identified a molecular circuit in which the expression of ciliary protein ADP-ribosylation factor-like protein 13B (ARL13B) is epigenetically regulated to promote adaptation to chemotherapy. Immuno-precipitation combined with liquid chromatography-mass spectrometry binding partner analysis revealed that that ARL13B interacts with the purine biosynthetic enzyme inosine-5'-monophosphate dehydrogenase 2 (IMPDH2). Further, radioisotope tracing revealed that this interaction functions as a negative regulator for purine salvaging. Inhibition of the ARL13B-IMPDH2 interaction enhances temozolomide-induced DNA damage by forcing glioblastoma cells to rely on the purine salvage pathway. Targeting the ARLI3B-IMPDH2 circuit can be achieved using the Food and Drug Administration-approved drug, mycophenolate mofetil, which can block IMPDH2 activity and enhance the therapeutic efficacy of temozolomide. Our results suggest and support clinical evaluation of MMF in combination with temozolomide treatment in glioma patients.

Keywords:  cellular plasticity; chemoresistance; glioblastoma; purine biosynthesis

DOI:  https://doi.org/10.1093/brain/awab020
Mol Cancer Res. 2021 Apr 16. pii: molcanres.0086.2020. [Epub ahead of print]

Elevated Asparagine Biosynthesis Drives Brain Tumor Stem Cell Metabolic Plasticity and Resistance to Oxidative Stress.

Tom M Thomas, Ken Miyaguchi, Lincoln A Edwards, Hongqiang Wang, Hassen Wollebo, Li Aiguo, Ramachandran Murali, Yizhou Wang, Daniel Braas, Justin S Michael, Allen Andres, Miqin Zhang, Kamel Khalili, Roberta A Gottlieb, J Manuel Perez, John S Yu.

Asparagine synthetase (ASNS) is a gene on the long arm of chromosome 7 that is copy number amplified in the majority of glioblastomas. ASNS copy number amplification is associated with a significantly decreased survival. Using patient-derived glioma stem cells (GSCs), we showed significant metabolic alterations occur in gliomas when perturbing the expression of asparagine synthetase, which is not merely restricted to amino acid homeostasis. ASNS-high GSCs maintained a slower basal metabolic profile yet readily shifted to a greatly increased capacity for glycolysis and oxidative phosphorylation when needed. This led ASNS-high cells to a greater ability to proliferate and spread into brain tissue. Finally, we demonstrate that these changes confer resistance to cellular stress, notably oxidative stress, through adaptive redox homeostasis which led to radiation resistance. Furthermore, ASNS overexpression led to modifications of the one-carbon metabolism to promote a more antioxidant tumor environment revealing a metabolic vulnerability that may be therapeutically exploited. Implications: This study reveals a new role for ASNS in metabolic control and redox homeostasis in glioma stem cells and proposes a new treatment strategy that attempts to exploit one vulnerable metabolic node within the larger multilayered tumor network.

DOI: https://doi.org/10.1158/1541-7786.MCR-20-0086
Brain. 2021 Apr 15. pii: awab155. [Epub ahead of print]

Tumor immune landscape of paediatric high-grade gliomas.

James L Ross, Jose Velazquez Vega, Ashley Plant, Tobey J MacDonald, Oren J Becher, Dolores Hambardzumyan.

  Over the last decade, remarkable progress has been made towards elucidating the origin and genomic landscape of childhood high-grade brain tumors. It has become evident that pediatric high-grade gliomas (pHGGs) differ from adult HGGs with respect to multiple defining aspects including: DNA copy number, gene expression profiles, tumor locations within the central nervous system, and genetic alterations such as somatic histone mutations. Despite these advances, clinical trials for children with glioma have historically been based on ineffective adult regimens that fail to take into consideration the fundamental biological differences between the two. Additionally, although our knowledge of the intrinsic cellular mechanisms driving tumor progression has considerably expanded, little is known concerning the dynamic tumor immune microenvironment (TIME) in pHGGs. In this review, we explore the genetic and epigenetic landscape of pHGGs and how this drives the creation of specific tumor sub-groups with meaningful survival outcomes. Further, we provide a comprehensive analysis of the pHGG TIME and discuss emerging therapeutic efforts aimed at exploiting the immune functions of these tumors.

Keywords:  Pediatric glioma; diffuse intrinsic pontine glioma; histone; inflammation; tumor associated macrophage

DOI:  https://doi.org/10.1093/brain/awab155
Proc Natl Acad Sci U S A. 2021 Apr 20. pii: e2009290118. [Epub ahead of print]118(16):

PI3Kγ inhibition suppresses microglia/TAM accumulation in glioblastoma microenvironment to promote exceptional temozolomide response.

Jie Li, Megan M Kaneda, Jun Ma, Ming Li, Ryan M Shepard, Kunal Patel, Tomoyuki Koga, Aaron Sarver, Frank Furnari, Beibei Xu, Sanjay Dhawan, Jianfang Ning, Hua Zhu, Anhua Wu, Gan You, Tao Jiang, Andrew S Venteicher, Jeremy N Rich, Christopher K Glass, Judith A Varner, Clark C Chen.

  Precision medicine in oncology leverages clinical observations of exceptional response. Toward an understanding of the molecular features that define this response, we applied an integrated, multiplatform analysis of RNA profiles derived from clinically annotated glioblastoma samples. This analysis suggested that specimens from exceptional responders are characterized by decreased accumulation of microglia/macrophages in the glioblastoma microenvironment. Glioblastoma-associated microglia/macrophages secreted interleukin 11 (IL11) to activate STAT3-MYC signaling in glioblastoma cells. This signaling induced stem cell states that confer enhanced tumorigenicity and resistance to the standard-of-care chemotherapy, temozolomide (TMZ). Targeting a myeloid cell restricted an isoform of phosphoinositide-3-kinase, phosphoinositide-3-kinase gamma isoform (PI3Kγ), by pharmacologic inhibition or genetic inactivation disrupted this signaling axis by reducing microglia/macrophage-associated IL11 secretion in the tumor microenvironment. Mirroring the clinical outcomes of exceptional responders, PI3Kγ inhibition synergistically enhanced the anti-neoplastic effects of TMZ in orthotopic murine glioblastoma models. Moreover, inhibition or genetic inactivation of PI3Kγ in murine glioblastoma models recapitulated expression profiles observed in clinical specimens isolated from exceptional responders. Our results suggest key contributions from tumor-associated microglia/macrophages in exceptional responses and highlight the translational potential for PI3Kγ inhibition as a glioblastoma therapy.

Keywords:  IL11; PI3Kγ; exceptional responders; glioblastoma; microglia/macrophages

DOI:  https://doi.org/10.1073/pnas.2009290118
Proc Natl Acad Sci U S A. 2021 Apr 20. pii: e2017148118. [Epub ahead of print]118(16):

Brd4-bound enhancers drive cell-intrinsic sex differences in glioblastoma.

Najla Kfoury, Zongtai Qi, Briana C Prager, Michael N Wilkinson, Lauren Broestl, Kristopher C Berrett, Arnav Moudgil, Sumithra Sankararaman, Xuhua Chen, Jason Gertz, Jeremy N Rich, Robi D Mitra, Joshua B Rubin.

  Sex can be an important determinant of cancer phenotype, and exploring sex-biased tumor biology holds promise for identifying novel therapeutic targets and new approaches to cancer treatment. In an established isogenic murine model of glioblastoma (GBM), we discovered correlated transcriptome-wide sex differences in gene expression, H3K27ac marks, large Brd4-bound enhancer usage, and Brd4 localization to Myc and p53 genomic binding sites. These sex-biased gene expression patterns were also evident in human glioblastoma stem cells (GSCs). These observations led us to hypothesize that Brd4-bound enhancers might underlie sex differences in stem cell function and tumorigenicity in GBM. We found that male and female GBM cells exhibited sex-specific responses to pharmacological or genetic inhibition of Brd4. Brd4 knockdown or pharmacologic inhibition decreased male GBM cell clonogenicity and in vivo tumorigenesis while increasing both in female GBM cells. These results were validated in male and female patient-derived GBM cell lines. Furthermore, analysis of the Cancer Therapeutic Response Portal of human GBM samples segregated by sex revealed that male GBM cells are significantly more sensitive to BET (bromodomain and extraterminal) inhibitors than are female cells. Thus, Brd4 activity is revealed to drive sex differences in stem cell and tumorigenic phenotypes, which can be abrogated by sex-specific responses to BET inhibition. This has important implications for the clinical evaluation and use of BET inhibitors.

Keywords:  BET inhibitors; Brd4-bound enhancers; glioblastoma; sex differences; sex-specific transcriptional programs

DOI:  https://doi.org/10.1073/pnas.2017148118
Acta Neuropathol Commun. 2021 Apr 14. 9(1): 69

Novel targetable FGFR2 and FGFR3 alterations in glioblastoma associate with aggressive phenotype and distinct gene expression programs.

Maria-Magdalena Georgescu, Mohammad Zahidul Islam, Yan Li, James Traylor, Anil Nanda.

  Prognostic molecular subgrouping of glioblastoma is an ongoing effort and the current classification includes IDH-wild-type and IDH-mutant entities, the latter showing significantly better prognosis. We performed a comparative integrated analysis of the FGFR glioblastoma subgroup consisting of 5 cases from a prospective 101-patient-cohort. FGFR alterations included FGFR2-TACC2 and FGFR2 amplifications arising in a multifocal IDH-mutant glioblastoma with unexpected 2.5-month patient survival, novel FGFR3 carboxy-terminal duplication and FGFR3-TLN1 fusion, and two previously described FGFR3-TACC3 fusions. The FGFR2 tumors showed additional mutations in SERPINE1/PAI-1 and MMP16, as part of extensive extracellular matrix remodeling programs. Whole transcriptomic analysis revealed common proliferation but distinct morphogenetic gene expression programs that correlated with tumor histology. The kinase program revealed EPHA3, LTK and ALK receptor tyrosine kinase overexpression in individual FGFR tumors. Paradoxically, all FGFR-fused glioblastomas shared strong PI3K and MAPK pathway suppression effected by SPRY, DUSP and AKAP12 inhibitors, whereas the FGFR2-TACC2 tumor elicited also EGFR suppression by ERRFI1 upregulation. This integrated analysis outlined the proliferation and morphogenetic expression programs in FGFR glioblastoma, and identified four novel, clinically targetable FGFR2 and FGFR3 alterations that confer aggressive phenotype and trigger canonical pathway feedback inhibition, with important therapeutic implications.

Keywords:  FGFR2 amplification; FGFR2-TACC2; FGFR3-TACC3; FGFR3-TLN1; Leptomeningeal gliomatosis; MMP; PAI-1; Proteomics; Transcriptomics

DOI:  https://doi.org/10.1186/s40478-021-01170-1
Clin Cancer Res. 2021 Apr 16. pii: clincanres.0446.2021. [Epub ahead of print]

Modified RANO (mRANO), iRANO, and standard RANO response to convection-enhanced delivery of IL4R-targeted immunotoxin MDNA55 in recurrent glioblastoma.

Benjamin M Ellingson, John H Sampson, Achal Singh Achrol, Manish K Aghi, Krystof Krystof Bankiewicz, Chencai Wang, Martin Bexon, Steven Brem, Andrew J Brenner, Sajeel Chowdhary, John R Floyd, Seunggu Jude Han, Santosh Kesari, Dina Randazzo, Michael A Vogelbaum, Frank D Vrionis, Miroslaw Zabek, Nicholas Butowski, Melissa Coello, Nina Merchant, Fahar Merchant.

PURPOSE: The current study compared the modified radiographic response assessment in neuro-oncology (mRANO), immunotherapy RANO (iRANO), and standard RANO criteria as well as quantified the association between progression-free (PFS) and overall survival (OS) in an immunotherapy trial in recurrent glioblastoma (rGBM).
EXPERIMENTAL DESIGN: 47 patients with rGBM were enrolled in a prospective phase II convection-enhanced delivery of an IL4R-targeted immunotoxin (MDNA55-05, NCT02858895). Bidirectional tumor measurements were created by local sites and centrally by an independent radiologic faculty (IRF), then standard RANO, iRANO, and mRANO criteria were applied.
RESULTS: 41 of 47 patients (mean age 56{plus minus}11.7) were evaluable for response. PFS was significantly shorter using standard RANO compared to iRANO (log-rank, P<0.0001; HR=0.3) and mRANO (P<0.0001;HR=0.3). In patients who died and had confirmed progression on standard RANO, no correlation was observed between PFS and OS (Local,P=0.47;Central,P=0.34). Using iRANO, a weak association was observed between confirmed PFS and OS via local site measurements (P=0.017), but not central measurements (P=0.18). 24 of 41 patients (59%) were censored using iRANO and because they lacked confirmation of progression 3 months after initial progression. A strong correlation was observed between mRANO PFS and OS for both local (R2 =0.66,P<0.0001) and centrally-determined reads (R2 =0.57,P=0.0007).
CONCLUSIONS: No correlation between radiographic PFS and OS was observed for standard RANO or iRANO, but a correlation was observed between PFS and OS using the mRANO criteria. Also, the iRANO criteria was difficult to implement due to need to confirm progression 3 months after initial progression, censoring more than half the patients.

DOI: https://doi.org/10.1158/1078-0432.CCR-21-0446
Cell Rep. 2021 Apr 13. pii: S2211-1247(21)00308-9. [Epub ahead of print]35(2): 108994

Dual targeting of the epigenome via FACT complex and histone deacetylase is a potent treatment strategy for DIPG.

Anahid Ehteda, Sandy Simon, Laura Franshaw, Federico M Giorgi, Jie Liu, Swapna Joshi, Jourdin R C Rouaen, Chi Nam Ignatius Pang, Ruby Pandher, Chelsea Mayoh, Yujie Tang, Aaminah Khan, Caitlin Ung, Ornella Tolhurst, Anne Kankean, Elisha Hayden, Rebecca Lehmann, Sylvie Shen, Anjana Gopalakrishnan, Peter Trebilcock, Katerina Gurova, Andrei V Gudkov, Murray D Norris, Michelle Haber, Orazio Vittorio, Maria Tsoli, David S Ziegler.

  Diffuse intrinsic pontine glioma (DIPG) is an aggressive and incurable childhood brain tumor for which new treatments are needed. CBL0137 is an anti-cancer compound developed from quinacrine that targets facilitates chromatin transcription (FACT), a chromatin remodeling complex involved in transcription, replication, and DNA repair. We show that CBL0137 displays profound cytotoxic activity against a panel of patient-derived DIPG cultures by restoring tumor suppressor TP53 and Rb activity. Moreover, in an orthotopic model of DIPG, treatment with CBL0137 significantly extends animal survival. The FACT subunit SPT16 is found to directly interact with H3.3K27M, and treatment with CBL0137 restores both histone H3 acetylation and trimethylation. Combined treatment of CBL0137 with the histone deacetylase inhibitor panobinostat leads to inhibition of the Rb/E2F1 pathway and induction of apoptosis. The combination of CBL0137 and panobinostat significantly prolongs the survival of mice bearing DIPG orthografts, suggesting a potential treatment strategy for DIPG.

Keywords:  DIPG; E2F1; EZH2; H3K27M; HDAC; anticancer therapy; brainstem glioma; facilitates chromatin transcription complex; pediatric cancer; xenograft model

DOI:  https://doi.org/10.1016/j.celrep.2021.108994
Neuro Oncol. 2021 Apr 17. pii: noab093. [Epub ahead of print]

Metabolic imaging detects elevated glucose flux through the pentose phosphate pathway associated with TERT expression in low-grade gliomas.

Pavithra Viswanath, Georgios Batsios, Vinay Ayyappan, Celiné Taglang, Anne Marie Gillespie, Peder E Z Larson, H Artee Luchman, Joseph F Costello, Russell O Pieper, Sabrina M Ronen.

   BACKGROUND: Telomerase reverse transcriptase (TERT) is essential for tumor proliferation, including in low-grade oligodendrogliomas (LGOGs). Since TERT is silenced in normal cells, it is also a therapeutic target. Therefore, non-invasive methods of imaging TERT are needed. Here, we examined the link between TERT expression and metabolism in LGOGs, with the goal of leveraging this information for non-invasive magnetic resonance spectroscopy (MRS)-based metabolic imaging of LGOGs.
METHODS: Immortalized normal human astrocytes with doxycycline-inducible TERT silencing, patient-derived LGOG cells, orthotopic tumors and LGOG patient biopsies were studied to determine the mechanistic link between TERT expression and glucose metabolism. The ability of hyperpolarized [U- 13C, U- 2H]-glucose to non-invasively assess TERT expression was tested in live cells and orthotopic tumors.
RESULTS: TERT expression was associated with elevated glucose flux through the pentose phosphate pathway (PPP), elevated NADPH, which is a major product of the PPP, and elevated GSH, which is maintained in a reduced state by NADPH. Importantly, hyperpolarized [U- 13C, U- 2H]-glucose metabolism via the PPP non-invasively reported on TERT expression and response to TERT inhibition in patient-derived LGOG cells and orthotopic tumors. Mechanistically, TERT acted via the sirtuin SIRT2 to upregulate the glucose transporter GLUT1 and the rate-limiting PPP enzyme glucose-6-phosphate dehydrogenase.
CONCLUSIONS: We have, for the first time, leveraged a mechanistic understanding of TERT-associated metabolic reprogramming for non-invasive imaging of LGOGs using hyperpolarized [U- 13C, U- 2H]-glucose. Our findings provide a novel way of imaging a hallmark of tumor immortality and have the potential to improve diagnosis and treatment response assessment for LGOG patients.

Keywords:  Telomerase; gliomas; glucose metabolism; hyperpolarized 13C; magnetic resonance spectroscopy

DOI:  https://doi.org/10.1093/neuonc/noab093
Neuro Oncol. 2021 Apr 17. pii: noab099. [Epub ahead of print]

Turning strength into weakness: protein degradation and autophagy as therapeutic targets in Glioblastoma?

Simone P Niclou, Anna Golebiewska.

DOI: https://doi.org/10.1093/neuonc/noab099
Cell Death Discov. 2021 Apr 16. 7(1): 81

Global phosphoproteomics reveals DYRK1A regulates CDK1 activity in glioblastoma cells.

Ariadna Recasens, Sean J Humphrey, Michael Ellis, Monira Hoque, Ramzi H Abbassi, Brianna Chen, Mitchell Longworth, Elise J Needham, David E James, Terrance G Johns, Bryan W Day, Michael Kassiou, Pengyi Yang, Lenka Munoz.

Both tumour suppressive and oncogenic functions have been reported for dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A). Herein, we performed a detailed investigation to delineate the role of DYRK1A in glioblastoma. Our phosphoproteomic and mechanistic studies show that DYRK1A induces degradation of cyclin B by phosphorylating CDC23, which is necessary for the function of the anaphase-promoting complex, a ubiquitin ligase that degrades mitotic proteins. DYRK1A inhibition leads to the accumulation of cyclin B and activation of CDK1. Importantly, we established that the phenotypic response of glioblastoma cells to DYRK1A inhibition depends on both retinoblastoma (RB) expression and the degree of residual DYRK1A activity. Moderate DYRK1A inhibition leads to moderate cyclin B accumulation, CDK1 activation and increased proliferation in RB-deficient cells. In RB-proficient cells, cyclin B/CDK1 activation in response to DYRK1A inhibition is neutralized by the RB pathway, resulting in an unchanged proliferation rate. In contrast, complete DYRK1A inhibition with high doses of inhibitors results in massive cyclin B accumulation, saturation of CDK1 activity and cell cycle arrest, regardless of RB status. These findings provide new insights into the complexity of context-dependent DYRK1A signalling in cancer cells.

DOI: https://doi.org/10.1038/s41420-021-00456-6
Cell. 2021 Apr 09. pii: S0092-8674(21)00351-2. [Epub ahead of print]

Glioblastomas acquire myeloid-affiliated transcriptional programs via epigenetic immunoediting to elicit immune evasion.

Ester Gangoso, Benjamin Southgate, Leanne Bradley, Stefanie Rus, Felipe Galvez-Cancino, Niamh McGivern, Esra Güç, Chantriolnt-Andreas Kapourani, Adam Byron, Kirsty M Ferguson, Neza Alfazema, Gillian Morrison, Vivien Grant, Carla Blin, IengFong Sou, Maria Angeles Marques-Torrejon, Lucia Conde, Simona Parrinello, Javier Herrero, Stephan Beck, Sebastian Brandner, Paul M Brennan, Paul Bertone, Jeffrey W Pollard, Sergio A Quezada, Duncan Sproul, Margaret C Frame, Alan Serrels, Steven M Pollard.

  Glioblastoma multiforme (GBM) is an aggressive brain tumor for which current immunotherapy approaches have been unsuccessful. Here, we explore the mechanisms underlying immune evasion in GBM. By serially transplanting GBM stem cells (GSCs) into immunocompetent hosts, we uncover an acquired capability of GSCs to escape immune clearance by establishing an enhanced immunosuppressive tumor microenvironment. Mechanistically, this is not elicited via genetic selection of tumor subclones, but through an epigenetic immunoediting process wherein stable transcriptional and epigenetic changes in GSCs are enforced following immune attack. These changes launch a myeloid-affiliated transcriptional program, which leads to increased recruitment of tumor-associated macrophages. Furthermore, we identify similar epigenetic and transcriptional signatures in human mesenchymal subtype GSCs. We conclude that epigenetic immunoediting may drive an acquired immune evasion program in the most aggressive mesenchymal GBM subtype by reshaping the tumor immune microenvironment.

Keywords:  DNA methylation; chemokine; epigenetics; glioblastoma; immune evasion; immunoediting; interferon signaling; macrophage; neural stem cell; syngeneic

DOI:  https://doi.org/10.1016/j.cell.2021.03.023
Neurooncol Adv. 2020 Jan-Dec;2(1):2(1): vdz056

Long-term survivors of glioblastoma are a unique group of patients lacking universal characteristic features.

Chul-Kee Park, Jeong Mo Bae, Sung-Hye Park.

DOI: https://doi.org/10.1093/noajnl/vdz056
Sci Rep. 2021 Apr 12. 11(1): 7927

Association of preoperative seizures with tumor metabolites quantified by magnetic resonance spectroscopy in gliomas.

Shunsuke Nakae, Masanobu Kumon, Kazuhiro Murayama, Shigeo Ohba, Hikaru Sasaki, Joji Inamasu, Kiyonori Kuwahara, Seiji Yamada, Masato Abe, Yuichi Hirose.

Seizures are common in patients with gliomas; however, the mechanisms of epileptogenesis in gliomas have not been fully understood. This study hypothesized that analyzing quantified metabolites using magnetic resonance spectroscopy (MRS) might provide novel insights to better understand the epileptogenesis in gliomas, and specific metabolites might be indicators of preoperative seizures in gliomas. We retrospectively investigated patient information (gender, age at diagnosis of tumor, their survival time) and tumor information (location, histology, genetic features, and metabolites according to MRS) in patients with gliomas. The data were correlated with the incidence of seizure and analyzed statistically. Of 146 adult supratentorial gliomas, isocitrate dehydrogenase (IDH) mutant tumors significantly indicated higher incidence of preoperative seizures than IDH wild-type gliomas. However, MRS study indicated that glutamate concentration in IDH wild-type gliomas was higher than that in IDH mutant gliomas. Glutamate was not associated with high frequency of preoperative seizures in patients with gliomas. Instead, increased total N-acetyl-L-aspartate (tNAA) was significantly associated with them. Moreover, multivariable analysis indicated that increased level of tNAA was an independent predictor of preoperative seizures. According to MRS analysis, tNAA, rather than glutamate, might be a useful to detect preoperative seizures in patient with supratentorial gliomas.

DOI: https://doi.org/10.1038/s41598-021-86487-6