Single-Cell RNA-Seq Analysis of Infiltrating Neoplastic Cells at the Migrating Front of Human Glioblastoma

Darmanis, Spyros; Sloan, Steven A.; Croote, Derek; Mignardi, Marco; Chernikova, Sophia B.; Samghababi, Peyman; Zhang, Ye; Neff, Norma; Kowarsky, Mark; Caneda, Christine; Li, Gordon; Chang, Steven D.; Connolly, Ian D.; Li, Yingmei; Barres, Ben A.; Gephart, Melanie Hayden; Quake, Stephen R.

doi:10.1016/j.celrep.2017.10.030

Cited by 748 publications

(969 citation statements)

References 31 publications

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“…Our data highlight the abundance of immunosuppressive pathways operating in GBM. The most abundant immune cells in GBM belong to the myeloid lineage , and we show that GSCs express cell surface molecules such as PD‐L1 and CD47, which inhibit phagocytosis by macrophage . These data demonstrate that immune inhibition within GBM is mediated by both immune and non‐immune lineages co‐operating to provide a pro‐tumour environment.…”

Section: Discussionmentioning

confidence: 67%

Expression profiling of single cells and patient cohorts identifies multiple immunosuppressive pathways and an altered NK cell phenotype in glioblastoma

Stead

Nsengimana

et al. 2019

Clinical and Experimental Immunology

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Summary Glioblastoma (GBM) is an aggressive cancer with a very poor prognosis. Generally viewed as weakly immunogenic, GBM responds poorly to current immunotherapies. To understand this problem more clearly we used a combination of natural killer (NK) cell functional assays together with gene and protein expression profiling to define the NK cell response to GBM and explore immunosuppression in the GBM microenvironment. In addition, we used transcriptome data from patient cohorts to classify GBM according to immunological profiles. We show that glioma stem‐like cells, a source of post‐treatment tumour recurrence, express multiple immunomodulatory cell surface molecules and are targeted in preference to normal neural progenitor cells by natural killer (NK) cells ex vivo. In contrast, GBM‐infiltrating NK cells express reduced levels of activation receptors within the tumour microenvironment, with hallmarks of transforming growth factor (TGF)‐β‐mediated inhibition. This NK cell inhibition is accompanied by expression of multiple immune checkpoint molecules on T cells. Single‐cell transcriptomics demonstrated that both tumour and haematopoietic‐derived cells in GBM express multiple, diverse mediators of immune evasion. Despite this, immunome analysis across a patient cohort identifies a spectrum of immunological activity in GBM, with active immunity marked by co‐expression of immune effector molecules and feedback inhibitory mechanisms. Our data show that GBM is recognized by the immune system but that anti‐tumour immunity is restrained by multiple immunosuppressive pathways, some of which operate in the healthy brain. The presence of immune activity in a subset of patients suggests that these patients will more probably benefit from combination immunotherapies directed against multiple immunosuppressive pathways.

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Section: Discussionmentioning

confidence: 67%

Expression profiling of single cells and patient cohorts identifies multiple immunosuppressive pathways and an altered NK cell phenotype in glioblastoma

Stead

Nsengimana

et al. 2019

Clinical and Experimental Immunology

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“…To validate whether the stem‐to‐invasion path could be recaptured in other GBM data, we obtained another single‐cell RNA‐seq data set of GBM (published by Darmanis et al () which contains 3589 cells from four patients. Following the same data processing, the final number of tumor cells for validation was 882.…”

Section: Resultsmentioning

confidence: 99%

Single‐cell RNA‐seq reveals the invasive trajectory and molecular cascades underlying glioblastoma progression

Pang

et al. 2019

Molecular Oncology

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Glioblastoma (GBM) is the most common and aggressive primary brain tumor, in which GBM stem cells (GSCs) were identified to contribute to aggressive phenotypes and poor prognosis. Yet, how GSCs progress to invasive cells remains largely unexplored. Here, we revealed the cell subpopulations with distinct functional status and the existence of cells with high invasive potential within heterogeneous primary GBM tumors. We reconstructed a branched trajectory by pseudotemporal ordering of single tumor cells, in which the root showed GSC‐like phenotype while the end displayed high invasive activity. Thus, we further determined a path along which GSCs gradually transformed to invasive cells, called the ‘stem‐to‐invasion path’. Along this path, cells showed incremental expression of GBM invasion‐associated signatures and diminishing expression of GBM stem cell markers. These findings were validated in an independent single‐cell data set of GBM. Through analyzing the molecular cascades underlying the path, we identify crucial factors controlling the attainment of invasive potential of tumor cells, including transcription factors and long noncoding RNAs. Our work provides novel insights into GBM progression, especially the attainment of invasive potential in primary tumor cells, and supports the cancer stem cell model, with valuable implications for GBM therapy.

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“…For single‐cell RNA sequencing analysis, normalized expression data along with metadata was retrieved from the companion webpage (http://www.gbmseq.org) associated with the original publication (Darmanis et al, ) on June 25, 2019. The data were subsequently filtered to retain only the top five upregulated genes from the astrocyte signature.…”

Section: Methodsmentioning

confidence: 99%

Astrocytes enhance glioblastoma growth

Mega

Nilsen

Leiss

et al. 2019

Glia

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Glioblastoma (GBM) is a deadly disease with a need for deeper understanding and new therapeutic approaches. The microenvironment of glioblastoma has previously been shown to guide glioblastoma progression. In this study, astrocytes were investigated with regard to their effect on glioblastoma proliferation through correlative analyses of clinical samples and experimental in vitro and in vivo studies. Co‐culture techniques were used to investigate the GBM growth enhancing potential of astrocytes. Cell sorting and RNA sequencing were used to generate a GBM‐associated astrocyte signature and to investigate astrocyte‐induced GBM genes. A NOD scid GBM mouse model was used for in vivo studies. A gene signature reflecting GBM‐activated astrocytes was associated with poor prognosis in the TCGA GBM dataset. Two genes, periostin and serglycin, induced in GBM cells upon exposure to astrocytes were expressed at higher levels in cases with high “astrocyte signature score”. Astrocytes were shown to enhance glioblastoma cell growth in cell lines and in a patient‐derived culture, in a manner dependent on cell–cell contact and involving increased cell proliferation. Furthermore, co‐injection of astrocytes with glioblastoma cells reduced survival in an orthotopic GBM model in NOD scid mice. In conclusion, this study suggests that astrocytes contribute to glioblastoma growth and implies this crosstalk as a candidate target for novel therapies.

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Single-Cell RNA-Seq Analysis of Infiltrating Neoplastic Cells at the Migrating Front of Human Glioblastoma

Cited by 748 publications

References 31 publications

Expression profiling of single cells and patient cohorts identifies multiple immunosuppressive pathways and an altered NK cell phenotype in glioblastoma

Expression profiling of single cells and patient cohorts identifies multiple immunosuppressive pathways and an altered NK cell phenotype in glioblastoma

Single‐cell RNA‐seq reveals the invasive trajectory and molecular cascades underlying glioblastoma progression

Astrocytes enhance glioblastoma growth

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