bims-malgli 2023-02-12 papers

bims-malgli

Biomed News

on Biology of malignant gliomas

Issue of 2023–02–12
eight papers selected by
Oltea Sampetrean, Keio University

Drugging a metabolic vulnerability in glioblastoma.
Preclinical Models of Low-Grade Gliomas.
Longitudinal DNA methylation analysis of adult-type IDH-mutant gliomas.
Tandem chimeric antigen receptor (CAR) T cells targeting EGFRvIII and IL-13Rα2 are effective against heterogeneous glioblastoma.
FGL2-targeting T cells exhibit antitumor effects on glioblastoma and recruit tumor-specific brain-resident memory T cells.
Integrative analysis of single-cell transcriptomics reveals age-associated immune landscape of glioblastoma.
Immunotherapy reverses glioma-driven dysfunction of immune system homeostasis.
Murine-Derived Glioma Organoids and Cell Line Culture Systems.

Nat Rev Drug Discov. 2023 Feb 07.

Drugging a metabolic vulnerability in glioblastoma.

Katie Kingwell.

DOI: https://doi.org/10.1038/d41573-023-00020-0
Cancers (Basel). 2023 Jan 18. pii: 596. [Epub ahead of print]15(3):

Preclinical Models of Low-Grade Gliomas.

Pushan Dasgupta, Veerakumar Balasubramanyian, John F de Groot, Nazanin K Majd.

  Diffuse infiltrating low-grade glioma (LGG) is classified as WHO grade 2 astrocytoma with isocitrate dehydrogenase (IDH) mutation and oligodendroglioma with IDH1 mutation and 1p/19q codeletion. Despite their better prognosis compared with glioblastoma, LGGs invariably recur, leading to disability and premature death. There is an unmet need to discover new therapeutics for LGG, which necessitates preclinical models that closely resemble the human disease. Basic scientific efforts in the field of neuro-oncology are mostly focused on high-grade glioma, due to the ease of maintaining rapidly growing cell cultures and highly reproducible murine tumors. Development of preclinical models of LGG, on the other hand, has been difficult due to the slow-growing nature of these tumors as well as challenges involved in recapitulating the widespread genomic and epigenomic effects of IDH mutation. The most recent WHO classification of CNS tumors emphasizes the importance of the role of IDH mutation in the classification of gliomas, yet there are relatively few IDH-mutant preclinical models available. Here, we review the in vitro and in vivo preclinical models of LGG and discuss the mechanistic challenges involved in generating such models and potential strategies to overcome these hurdles.

Keywords:  IDH-mutant glioma; glioma; low-grade glioma; patient avatars; preclinical models

DOI:  https://doi.org/10.3390/cancers15030596
Acta Neuropathol Commun. 2023 Feb 04. 11(1): 23

Longitudinal DNA methylation analysis of adult-type IDH-mutant gliomas.

Sandra Ferreyra Vega, Thomas Olsson Bontell, Teresia Kling, Asgeir Store Jakola, Helena Carén.

  Diffuse gliomas are the most prevalent malignant primary brain tumors in adults and remain incurable despite standard therapy. Tumor recurrence is currently inevitable, which contributes to a persistent high morbidity and mortality in these patients. In this study, we examined the genome-wide DNA methylation profiles of primary and recurrent adult-type IDH-mutant gliomas to elucidate DNA methylation changes associated with tumor progression (with or without malignant transformation). We analyzed DNA methylation profiles of 37 primary IDH-mutant gliomas and 42 paired recurrences using the DNA methylation EPIC beadChip array. DNA methylation-based classification reflected the tumor progression over time. We observed a methylation subtype switch in a proportion of IDH-mutant astrocytomas; the primary tumors were subclassified as low-grade astrocytomas, which progressed to high-grade astrocytomas in the recurrent tumors. The CNS WHO grade 4 IDH-mutant astrocytomas did not always resemble methylation subclasses of higher grades. The number of differentially methylated CpG sites increased over time, and astrocytomas accumulated more differentially methylated CpG sites than oligodendrogliomas during tumor progression. Few differentially methylated CpG sites were shared between patients. We demonstrated that DNA methylation profiles are mostly maintained during IDH-mutant glioma progression, but CpG site-specific methylation alterations can occur.

Keywords:  DNA methylation profiling; IDH-mutant gliomas; Longitudinal analysis; Tumor recurrence

DOI:  https://doi.org/10.1186/s40478-023-01520-1
Neurooncol Adv. 2023 Jan-Dec;5(1):5(1): vdac185

Tandem chimeric antigen receptor (CAR) T cells targeting EGFRvIII and IL-13Rα2 are effective against heterogeneous glioblastoma.

Andrea Schmidts, Ambike A Srivastava, Rishab Ramapriyan, Stefanie R Bailey, Amanda A Bouffard, Daniel P Cahill, Bob S Carter, William T Curry, Gavin P Dunn, Matthew J Frigault, Elizabeth R Gerstner, Jack Y Ghannam, Michael C Kann, Rebecca C Larson, Mark B Leick, Brian V Nahed, Leland G Richardson, Irene Scarfò, Jing Sun, Hiroaki Wakimoto, Marcela V Maus, Bryan D Choi.

   Background: Chimeric antigen receptor (CAR) T cells have achieved remarkable responses in patients with hematological malignancies; however, the potential of this therapeutic platform for solid tumors like glioblastoma (GBM) has been limited, due in large part to the targeting of single antigens in a heterogeneous disease. Strategies that allow CAR T cells to engage multiple antigens concomitantly may broaden therapeutic responses and mitigate the effects of immune escape.
Methods: Here we have developed a novel, dual-specific, tandem CAR T (TanCART) cell with the ability to simultaneously target both EGFRvIII and IL-13Rα2, two well-characterized tumor antigens that are frequently found on the surface of GBM cells but completely absent from normal brain tissues. We employed both standard immunological assays and multiple orthotopic preclinical models including patient-derived xenograft to demonstrate efficacy of this approach against heterogeneous tumors.
Results: Tandem CAR T cells displayed enhanced cytotoxicity in vitro against heterogeneous GBM populations, including patient-derived brain tumor cultures (P < .05). Compared to CAR T cells targeting single antigens, dual antigen engagement through the tandem construct was necessary to achieve long-term, complete, and durable responses in orthotopic murine models of heterogeneous GBM, including patient-derived xenografts (P < .05).
Conclusions: We demonstrate that TanCART is effective against heterogeneous tumors in the brain. These data lend further credence to the development of multi-specific CAR T cells in the treatment of GBM and other cancers.

Keywords:  chimeric antigen receptor; glioblastoma; immunotherapy

DOI:  https://doi.org/10.1093/noajnl/vdac185
Nat Commun. 2023 Feb 10. 14(1): 735

FGL2-targeting T cells exhibit antitumor effects on glioblastoma and recruit tumor-specific brain-resident memory T cells.

Qingnan Zhao, Jiemiao Hu, Lingyuan Kong, Shan Jiang, Xiangjun Tian, Jing Wang, Rintaro Hashizume, Zhiliang Jia, Natalie Wall Fowlkes, Jun Yan, Xueqing Xia, Sofia F Yi, Long Hoang Dao, David Masopust, Amy B Heimberger, Shulin Li.

Although tissue-resident memory T (TRM) cells specific for previously encountered pathogens have been characterized, the induction and recruitment of brain TRM cells following immune therapy has not been observed in the context of glioblastoma. Here, we show that T cells expressing fibrinogen-like 2 (FGL2)-specific single-chain variable fragments (T-αFGL2) can induce tumor-specific CD8+ TRM cells that prevent glioblastoma recurrence. These CD8+ TRM cells display a highly expanded T cell receptor repertoire distinct from that found in peripheral tissue. When adoptively transferred to the brains of either immunocompetent or T cell-deficient naïve mice, these CD8+ TRM cells reject glioma cells. Mechanistically, T-αFGL2 cell treatment increased the number of CD69+CD8+ brain-resident memory T cells in tumor-bearing mice via a CXCL9/10 and CXCR3 chemokine axis. These findings suggest that tumor-specific brain-resident CD8+ TRM cells may have promising implications for the prevention of brain tumor recurrence.

DOI: https://doi.org/10.1038/s41467-023-36430-2
Front Immunol. 2023 ;14 1028775

Integrative analysis of single-cell transcriptomics reveals age-associated immune landscape of glioblastoma.

Songang Wu, Xuewen Li, Fan Hong, Qiang Chen, Yingying Yu, Shuanghui Guo, Yuanyuan Xie, Naian Xiao, Xuwen Kong, Wei Mo, Zhanxiang Wang, Shaoxuan Chen, Feng Zeng.

  Glioblastoma (GBM) is the most malignant tumor in center nervous system. Clinical statistics revealed that senior GBM patients had a worse overall survival (OS) comparing with that of patients in other ages, which is mainly related with tumor microenvironment including tumor-associated immune cells in particular. However, the immune heterogeneity and age-related prognosis in GBM are under studied. Here we developed a machine learning-based method to integrate public large-scale single-cell RNA sequencing (scRNA-seq) datasets to establish a comprehensive atlas of immune cells infiltrating in cross-age GBM. We found that the compositions of the immune cells are remarkably different across ages. Brain-resident microglia constitute the majority of glioblastoma-associated macrophages (GAMs) in patients, whereas dramatic elevation of extracranial monocyte-derived macrophages (MDMs) is observed in GAMs of senior patients, which contributes to the worse prognosis of aged patients. Further analysis suggests that the increased MDMs arisen from excessive recruitment and proliferation of peripheral monocytes not only lead to the T cell function inhibition in GBM, but also stimulate tumor cells proliferation via VEGFA secretion. In summary, our work provides new cues for the correlational relationship between the immune microenvironment of GBM and aging, which might be insightful for precise and effective therapeutic interventions for senior GBM patients.

Keywords:  GBM; age; immune microenvironment; monocyte-derived macrophage (MDM); single-cell RNA sequencing (scRNA-seq); tumor heterogeneity

DOI:  https://doi.org/10.3389/fimmu.2023.1028775
J Immunother Cancer. 2023 Feb;pii: e004805. [Epub ahead of print]11(2):

Immunotherapy reverses glioma-driven dysfunction of immune system homeostasis.

Bayli DiVita Dean, Tyler Wildes, Joseph Dean, Oleg Yegorov, Changlin Yang, David Shin, Connor Francis, John W Figg, Mathew Sebastian, Laura Falceto Font, Dan Jin, Alexandra Reid, Ginger Moore, Brandon Fernandez, Brandon Wummer, Carmelle Kuizon, Duane Mitchell, Catherine T Flores.

   BACKGROUND: Glioma-induced immune dysregulation of the hematopoietic system has been described in a limited number of studies. In this study, our group further demonstrates that gliomas interrupt the cellular differentiation programming and outcomes of hematopoietic stem and progenitor cells (HSPCs) in the bone marrow. HSPCs from glioma-bearing mice are reprogrammed and driven towards expansion of myeloid lineage precursors and myeloid-derived suppressor cells (MDSCs) in secondary lymphoid organs. However, we found this expansion is reversed by immunotherapy. Adoptive cellular therapy (ACT) has been demonstrably efficacious in multiple preclinical models of central nervous system (CNS) malignancies, and here we describe how glioma-induced dysfunction is reversed by this immunotherapeutic platform.
METHODS: The impact of orthotopic KR158B-luc glioma on HSPCs was evaluated in an unbiased fashion using single cell RNAseq (scRNAseq) of lineage- cells and phenotypically using flow cytometry. Mature myeloid cell frequencies and function were also evaluated using flow cytometry. Finally, ACT containing total body irradiation, tumor RNA-pulsed dendritic cells, tumor-reactive T cells and HSPCs isolated from glioma-bearing or non-tumor-bearing mice were used to evaluate cell fate differentiation and survival.
RESULTS: Using scRNAseq, we observed an altered HSPC landscape in glioma-bearing versus non-tumor-bearing mice . In addition, an expansion of myeloid lineage subsets, including granulocyte macrophage precursors (GMPs) and MDSCs, were observed in glioma-bearing mice relative to non-tumor-bearing controls. Furthermore, MDSCs from glioma-bearing mice demonstrated increased suppressive capacity toward tumor-specific T cells as compared with MDSCs from non-tumor-bearing hosts. Interestingly, treatment with ACT overcame these suppressive properties. When HSPCs from glioma-bearing mice were transferred in the context of ACT, we observed significant survival benefit and long-term cures in orthotopic glioma models compared with mice treated with ACT using non-glioma-bearing HSPCs.

Keywords:  dendritic cells; immunotherapy; myeloid-derived suppressor cells

DOI:  https://doi.org/10.1136/jitc-2022-004805
Curr Protoc. 2023 Feb;3(2): e665

Murine-Derived Glioma Organoids and Cell Line Culture Systems.

Alberto E Ayala-Sarmiento, David Rincon Fernandez Pacheco, Joshua J Breunig.

  Research models in cancer have greatly evolved in the last decade, with the advent of several new methods both in vitro and in vivo. While in vivo models remain the gold standard for preclinical studies, these methods present a series of disadvantages such as a high cost and long periods of time to produce results compared with in vitro models. We have previously developed a method named Mosaic Analysis by Dual Recombinase-mediated cassette exchange (MADR) that generates autochthonous gliomas in immunocompetent mice through the transgenesis of personalized driver mutations, which highly mimic the spatial and temporal tumor development of their human counterparts. Due to the control of single-copy expression of transgenes, it allows for comparing the visualization of tumor cells and non-tumor cells. Here we describe a method to generate murine-derived glioma organoids (MGOs) and cell line cultures from these murine models by physical and enzymatic methods for in vitro downstream applications. Tumor cells can be readily distinguished from non-tumor cell populations, in both organoids and monolayer cell cultures, and isolated due to the use of personalized fluorescent reporter transgenes. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Generation of 3D murine-derived glioma organoids Basic Protocol 2: Generation of 2D glioma monolayer cell lines.

Keywords:  MADR; biobank; glioma; organoids; personalized driver mutations

DOI:  https://doi.org/10.1002/cpz1.665