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:Here we performed a ChIP-seq experiment for Zeb1 trancription factor on a sample of adherent cultures of human neural stem cells (Cb192 cell line) and of a human glioblastoma cancer stem-like cell line (NCH421k). The result is the generation of the genome-wide maps for Zeb1 binding to chromatin in human neural stem cells and glioblastoma stem-like cells.
Project description:With HiRIEF LC-MS/MS shotgun proteomics, we analysed 6 patient-derived glioblastoma stem cells (BT stem cells) and compared them to an astrocyte line (CliniSciences, Guidonia Montecelio, Italy) and a more differentiated glioblastoma cell line (T98G). Each of the 8 cell line sample was run in triplicates in a total of three TMT10 sets, assigning each replicate in a separate TMT10 set, using 2 internal standards per set. The three TMT10 sets were ran in two experiments, first on immobilized pH gradient (IPG) 3-10 isoelectric focusing (IEF) strips, and then on IPG 3.7-4.9 IEF strips.
Project description:m6A RNA methylation plays an important role in regulating the self-renewal and tumorigenesis of glioblastoma stem cells. We performed m6A-seq analysis in glioblastoma stem cells with m6A levels modulated by knocking down METTL3 or METTL14 and identified m6A regulated genes in glioblastoma stem cells.
Project description:Chromatin accessibility discriminates stem from mature cell populations, enabling the identification of primitive stem-like cells in primary tumors, such as Glioblastoma (GBM) where self-renewing cells driving cancer progression and recurrence are prime targets for therapeutic intervention. We show, using single-cell chromatin accessibility, that primary GBMs harbor a heterogeneous self-renewing population whose diversity is captured in patient-derived glioblastoma stem cells (GSCs). In depth characterization of chromatin accessibility in GSCs identifies three GSC states: Reactive, Constructive, and Invasive, each governed by uniquely essential transcription factors and present within GBMs in varying proportions. Orthotopic xenografts reveal that GSC states associate with survival, and identify an invasive GSC signature predictive of low patient survival. Our chromatin-driven characterization of GSC states improves prognostic precision and identifies dependencies to guide combination therapies.
