Project description:Glioma stem cells (GSCs) have been identified in glioma tissues and suggested to play important roles in the tumorigenesis of glioblastoma multiform (GBM). We established a novel cellular bioinformative pipeline that consisted of principal component analysis (PCA) with factor loading, intracellular pathway analysis, and immunopathway analysis and attempted to clarify the differences in gene expression profiles comprehensively among GSCs, a glioma cell line (U251), and a human GBM tissue (hGBM). To this end, we extracted total RNAs from the GSCs, U251, and the hGBM, performed microarray, and applied the data to the bioinformatics analyses described above. As the results, PCA clearly distinguished the three groups. Moreover, the second principal component (PC2) distinguished the GSCs from the hGBM and U251; it reflected the characteristics of stemness. The factor loading for PC2 suggested MYCN, DPP4, and, MIF as contributing factors to the stemness of GSCs. We clarify the similarities and differences among samples such as the GSCs, U251, and hGBM.
Project description:We compared a large panel of human glioblastoma stem-like (GS) cell lines, corresponding primary tumors and conventional glioma cell lines to identify cell lines that preserve the transcriptome of human glioblastomas most closely, thereby allowing identification of shared therapeutic targets. We used Affymetrix HG-U133 Plus 2.0 microarrays to compare human glioblastoma stem-like (GS) cell lines, corresponding primary tumors and conventional glioma cell lines. We extracted total RNA from 32 conventional glioma cell lines, 12 GS cell lines (8 in two different passages), 7 clonal sublines derived from two GS lines, 12 original tumors, and 4 monolayer cultures established from the same tumors as GS-lines using standard serum conditions.
Project description:We compared a large panel of human glioblastoma stem-like (GS) cell lines, corresponding primary tumors and conventional glioma cell lines to identify cell lines that preserve the transcriptome of human glioblastomas most closely, thereby allowing identification of shared therapeutic targets. We used Affymetrix HG-U133 Plus 2.0 microarrays to compare human glioblastoma stem-like (GS) cell lines, corresponding primary tumors and conventional glioma cell lines.
Project description:Glioblastoma is one of the most malignant brain tumors with poor prognosis and their development and progression are known to be driven by glioblastoma stem cells. Although glioblastoma stem cells lose their cancer stemness properties during cultivation in serum-containing medium, little is known about the molecular mechanisms regulating signaling alteration in relation to reduction of stemness. In order to elucidate the global phosphorylation-related signaling events, we performed a SILAC-based quantitative phosphoproteome analysis of serum-induced dynamics in glioblastoma stem cells established from the tumor tissues of the patient. Among a total of 2,876 phosphorylation sites on 1,584 proteins identified in our analysis, 732 phosphorylation sites on 419 proteins were regulated through the alteration of stem cell characteristics.
Project description:Piwi proteins are a subfamily of Argonaute proteins that maintain germ cells in eukaryotes. However, the role of their human homologues in cancer stem cells and more broadly in cancer is poorly understood. Here, we report that the Piwi-like family members, including Piwil1 (Hiwi), are overexpressed in glioblastoma (GBM), with Piwil1 levels most frequently elevated. Piwil1 is enriched in glioma stem cells (GSCs) and helps to maintain their self-renewal. GSCs were transduces with control non-targeting shRNAs (shNT) and shPiwil1 (#1 and #2) and global gene expression was analyzed to identify Piwil1 downscream singalings.
Project description:Glioblastomas are the most lethal tumors affecting the central nervous system in adults. Simple and inexpensive syngeneic in vivo models that closely mirror human glioblastoma, including interactions between tumor and immune cells, are urgently needed for deciphering glioma biology and developing more effective treatments. Here, we generated mouse glioblastoma cell lines by repeated in-vivo passaging of neural stem cells and tumor tissue of a neural stem cell-specific Pten/p53 double-knockout genetic mouse model. Transcriptome and genome analyses of the cell lines revealed molecular heterogeneity comparable to that observed in human glioblastoma. Upon orthotopic transplantation into syngeneic hosts they formed high-grade gliomas that faithfully recapitulated the histopathological characteristics, invasiveness and infiltration by myeloid cells characteristic of human glioblastoma. These features make our cell lines unique and useful tools to study multiple aspects of glioma pathomechanism and test immunotherapies in syngeneic preclinical models.
Project description:Glioblastomas are the most lethal tumors affecting the central nervous system in adults. Simple and inexpensive syngeneic in vivo models that closely mirror human glioblastoma, including interactions between tumor and immune cells, are urgently needed for deciphering glioma biology and developing more effective treatments. Here, we generated mouse glioblastoma cell lines by repeated in-vivo passaging of neural stem cells and tumor tissue of a neural stem cell-specific Pten/p53 double-knockout genetic mouse model. Transcriptome and genome analyses of the cell lines revealed molecular heterogeneity comparable to that observed in human glioblastoma. Upon orthotopic transplantation into syngeneic hosts they formed high-grade gliomas that faithfully recapitulated the histopathological characteristics, invasiveness and infiltration by myeloid cells characteristic of human glioblastoma. These features make our cell lines unique and useful tools to study multiple aspects of glioma pathomechanism and test immunotherapies in syngeneic preclinical models.
Project description:Glioblastoma, the most common and malignant brain tumor, harbors stem-like cells that self-renew and propagate upon serial transplantation. Although they share functional, morphological and developmental similarities to adult brain neural stem cells, stem cell characteristic pathways contributing to glioblastoma stem-like cells have not been consistently determined. Towards this goal we have provided an internally coherent molecular reference that compares adult neural and glioblastoma stem cells cultivated under identical conditions. Genes dysregulated between these populations are correlated with clinical outcome in glioblastoma, and are highly expressed in embryonic and induced pluripotent stem cells. The resource yields a key role for Wnt and a core group of dysregulated pathways. We have verified the contribution to proliferation and sphere formation of Wnt- differentially activated genes through the Wnt inhibitor SFRP1. The glioblastoma and neural stem cell gene-expression and pathway comparative resource provides a powerful new tool for the identification of potential therapeutic targets. 14 samples was analyzed; 5 individual samples of adult neural stem cells and 9 individual samples of glioma stem cells
Project description:Glioma initiating cells/stem cells exist in the bulk tumor of glioblastoma. This cell population contributes to the frequent resistances toward radiation/chemotherapy, aggressiveness of adult brain cancer and increased recurrence rate. Targeting stem cell population becomes one the most promising and permissive therapeutic strategies. We isolated glioma stem cells from patient-derived xenografts and profiled their epigenomic features, including 4 different DNA marks and 2 enhancer marks, and transcriptome in these in vitro cultured cell lines. Three fetal brain-derived neural stem/progenitors cells were used for comparing the unique and common molecular features in these glioma cancer stem cells.