Glioblastoma stem cells reprogram chromatin in vivo to generate selective therapeutic dependencies on DPY30 and phosphodiesterases

2022 ◽  
Vol 14 (626) ◽  
Author(s):  
Deobrat Dixit ◽  
Briana C. Prager ◽  
Ryan C. Gimple ◽  
Tyler E. Miller ◽  
Qiulian Wu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Glioblastoma Stem Cells

scholarly journals OMRT-11. The effect of microenvironment on glioblastoma stem cells therapeutic resistance

2021 ◽  
Vol 3 (Supplement_2) ◽  
pp. ii9-ii9
Author(s):  
Tamara Lah Turnsek ◽  
Barbara Breznik ◽  
Bernarda Majc ◽  
Metka Novak ◽  
Andrej Porčnik ◽  
...  
Keyword(s):  
Stem Cells ◽  
Epithelial To Mesenchymal Transition ◽  
Combined Treatment ◽  
Cell Plasticity ◽  
Glioblastoma Stem Cells ◽  
Mesenchymal Transition ◽  
Non Invasive ◽  
Differential Gene

Abstract Epithelial-to-mesenchymal transition (EMT) is an essential molecular and cellular process in physiologic processes and invasion of various types of carcinoma and glioblastoma (GBM) cells. EMT is activated and regulated by specific endogenous triggers in complex network of intercellular interactions and signaling pathways. The hallmark of cancer-linked EMT are intermediate states that show notable cell plasticity, characteristic of cancer stem cells (CSCs), including glioblastoma stem cells – GSCs. GSCs resistance to irradiation (IR) and temozolomide (TMZ) chemotherapy is responsible for early relapses, even at distant brain sites. As GSCs are mostly homing to their “niches” as slowly-dividing GSC-subtype, mimicking a proneural-like non- invasive phenotype PN-genotype, we assume that this, by undergoing an EMT-like transition, GSCs are-reprogrammed to an invasive mesenchymal (MES) GBs/GSCs phenotype in a processes, called PMT (1). However, it is not known, if and by which environmental cues within the niche, this transition of GSCs is induced in vivo. In this work, we are presenting the transriptome data obtained when we exposed GSC spheroids to irradiation alone, TMZ alone and to the combined treatment in vitro and compared their differential genetic fingerprints related to EMT/PMT transition to the GSCs PMT transition, when embedded in their natural microenvironment in the GBM organoid model. The differential gene expression upon GSCs therapeutic perturbation (when alone and vs in the tumoroid microenvironment) will reveal the effects of the major candidate genes, associated with micronevironmendt stromal cells and matrix are contributing their observed EMT/PMT transition of GSCs in vivo. •1. Majc, B., Sever, T., Zarić, M, Breznik, B., Turk, B, Lah Turnšek, T. Epithelial- to-mesenchymal transition as the driver of changing carcinoma and glioblastoma microenvironment. DOI: 10.1016/j.bbamcr.2020.118782

scholarly journals STEM-12. ZIKA VIRUS TARGETS GLIOBLASTOMA STEM CELLS THROUGH A SOX2-INTEGRIN α vβ 5 AXIS

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii198-ii199
Author(s):  
Clark Chen ◽  
Sanjay Dhawan ◽  
Zhe Zhu ◽  
Pinar Mesci ◽  
Jeremy Rich
Keyword(s):  
Stem Cells ◽  
Zika Virus ◽  
Oncolytic Virus ◽  
Great Promise ◽  
Normal Brain ◽  
Zikv Infection ◽  
Glioblastoma Stem Cells ◽  
Interferon Stimulated Genes ◽  
Sox2 Expression

Abstract INTRODUCTION Oncolytic virus hold great promise as a platform for glioblastoma therapeutic development. Zika virus (ZIKV) is an oncolytic virus with exquisite selectivity for infecting and killing glioblastoma stem cells (GSCs). Here, we delineate the molecular determinant of this selectivity. METHODS cell-based glioblastoma models, glioblastoma organoid assays, in vivo murine glioblastoma models, ZIKV infectivity assays, gene silencing, ChIP-seq studies. RESULTS In independent models, ZIKV preferentially infected and lysed SOX2+ GSCs. Silencing of SOX2 expression attenuated this preferential infectivity. Of note, ZIKV infection of GSCs was independent of AXL, its putative receptor in normal brain. ChIP-seq experiments revealed that SOX2 bound within the ITGAV locus (encoding the integrin av subunit), and this binding was associated with accumulation of the active chromatin mark H3K27ac. Silencing of SOX2 suppressed ITGAV expression as well as ZIKV infectivity against GSCs, indicating that integrin is required for ZIKV infection. Of integrin b units, only silencing of integrin b5 prevented the killing of GSCs by ZIKV infection, suggesting ZIKV infection required the avb5 integrin. Supporting this hypothesis, blockade of the avb5 integrin substantially reduced ZIKV infection of GSCs in glioblastoma organoid assays and in clinical glioblastoma specimens. Sox2 expression additionally suppress GSC expression of all members of the interferon-stimulated genes (ISG family), thereby suppressing innate anti-viral response to facilitate ZIKV infection. CONCLUSIONS Collectively, our results reveal that ZIKV infection of GSCs is mediated by integrin α vβ 5 leading to SOX2 expression which negatively regulates antiviral immunity thereby facilitating ZIKV infection.

scholarly journals Isoflurane enhances the malignant potential of glioblastoma stem cells by promoting their viability, mobility in vitro and migratory capacity in vivo

2016 ◽  
Vol 116 (6) ◽  
pp. 870-877 ◽  
Author(s):  
M Zhu ◽  
M Li ◽  
Y Zhou ◽  
S Dangelmajer ◽  
U.D. Kahlert ◽  
...  
Keyword(s):  
Stem Cells ◽  
Malignant Potential ◽  
Glioblastoma Stem Cells ◽  
Migratory Capacity

scholarly journals PP54. NANO-ENCAPSULATED DISULFIRAM TARGETS GLIOBLASTOMA STEM CELLS IN VITRO AND IN VIVO BY MODULATION OF HYPOXIA-NF-KB PATHWAY

2017 ◽  
Vol 19 (suppl_1) ◽  
pp. i15-i15
Author(s):  
Dr Vinodh Kannappan ◽  
Dr Zhipeng Wang ◽  
Dr Peng Liu ◽  
Dr Sarah Brown ◽  
Ms Kate Butcher ◽  
...  
Keyword(s):  
Stem Cells ◽  
Glioblastoma Stem Cells

Demethoxycurcumin was superior to temozolomide in the inhibition of the growth of glioblastoma stem cells in vivo

Tumor Biology ◽  
2016 ◽  
Vol 37 (12) ◽  
pp. 15847-15857 ◽  
Author(s):  
Liang Leng ◽  
Xiaojun Zhong ◽  
Guan Sun ◽  
Wen Qiu ◽  
Lei Shi
Keyword(s):  
Stem Cells ◽  
Glioblastoma Stem Cells

scholarly journals TMIC-22. IDENTIFICATION OF CANCER-ASSOCIATED FIBROBLASTS IN GLIOBLASTOMA and Defining Their Protumoral Effects

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi252-vi252
Author(s):  
Jonathan Rick ◽  
Alan Nguyen ◽  
Ankush Chandra ◽  
Harsh Wadhwa ◽  
Sumedh Shah ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Stem Cell ◽  
Cancer Associated Fibroblasts ◽  
Rna Seq ◽  
Glioblastoma Stem Cells ◽  
Perivascular Niche ◽  
Glioblastoma Stem Cell ◽  
Blocking Antibodies

Abstract While cancer-associated fibroblasts (CAFs) and their pro-tumoral effects have been demonstrated in systemic cancers, CAFs had been presumed absent in glioblastoma given the lack of normal fibroblasts in the healthy brain. Here, we show that 5–26% (mean=12%) of cells in human glioblastomas express CAF markers α-SMA or PDGFR-β, morphologically resemble fibroblasts, and transcriptomically resemble by RNA-seq CAFs from other cancers. Glioblastoma CAFs were chemotactically attracted to glioblastoma-initiating stem cells (P=0.02). While glioblastoma CAFs did not affect differentiated glioblastoma cell proliferation (P=0.4), CAFs increased glioblastoma stem cell proliferation (P=0.002) and expression of glioblastoma stem cell-associated genes (P< 0.001). To identify mediators of CAF/glioblastoma stem cell interactions, we created a resource of inferred crosstalk by mapping the expression of receptors to that of their cognate ligands/agonists, using our RNA-seq results from glioblastoma CAFs and stem cells, revealing PDGF-β/PDGFR and osteopontin/CD44 to mediate stem cell recruitment of CAFs and CAF enrichment of stem cells, as confirmed by blocking antibodies (P=0.02–0.03). CAFs also render the glioblastoma microenvironment more pro-tumoral by promoting M2 polarization of tumor-associated macrophages (P=0.01), an effect we found to arise from unique CAF production of the EDA splice variant of fibronectin binding toll-like receptor 4 (TLR4), a known EDA receptor expressed by macrophages (P=0.02). In patient glioblastomas, CAFs were enriched 3-fold in the subventricular zone (SVZ) (P=0.04) which houses the neural stem cells that generate glioblastoma stem cells. SVZs from epilepsy cases or autopsies of glioblastoma-containing brains without ventricular involvement lacked CAFs. Depleting CAFs in xenografts derived from neurosphere-containing glioblastoma stem cells slowed their growth in vivo (P< 0.001). These findings are among the first to identify and profile glioblastoma CAFs. CAF recruitment by glioblastoma stem cells and creation of a pro-tumoral microenvironment in the perivascular niche housing glioblastoma stem cells, particularly in the SVZ, makes them an intriguing therapeutic target.

scholarly journals STEM-22. TARGETING PYRIMIDINE SYNTHESIS ACCENTUATES MOLECULAR THERAPY RESPONSE IN GLIOBLASTOMA STEM CELLS

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi238-vi238
Author(s):  
Kailin Yang ◽  
Xiuxing Wang ◽  
Qiulian Wu ◽  
Leo Kim ◽  
Andrew Morton ◽  
...  
Keyword(s):  
Stem Cells ◽  
De Novo ◽  
Metabolic Reprogramming ◽  
Molecular Therapy ◽  
Driver Mutations ◽  
Carbamoyl Phosphate ◽  
Glioblastoma Stem Cells ◽  
Pyrimidine Synthesis ◽  
Pyrimidine Nucleotide

Abstract Glioblastoma stem cells (GSCs) reprogram glucose metabolism by hijacking high-affinity glucose uptake to survive in a nutritionally dynamic microenvironment. Here, we trace metabolic aberrations in GSCs to link core genetic mutations in glioblastoma to dependency on de novo pyrimidine synthesis. Targeting the pyrimidine synthetic rate-limiting step enzyme carbamoyl-phosphate synthetase 2, aspartate transcarbamyolase, dihydroorotase (CAD) or the critical downstream enzyme, dihydroorotate dehydrogenase (DHODH) inhibited GSC survival, self-renewal, and in vivo tumor initiation through the depletion of the pyrimidine nucleotide supply in rodent models. Mutations in EGFR or PTEN generated distinct CAD phosphorylation patterns to activate carbon influx through pyrimidine synthesis. Simultaneous abrogation of tumor-specific driver mutations and DHODH activity with clinically approved inhibitors demonstrated sustained inhibition of metabolic activity of pyrimidine synthesis and GSC tumorigenic capacity. Higher expression of pyrimidine synthesis genes portend poor prognosis of glioblastoma patients. Collectively, our results demonstrate a therapeutic approach of precision medicine through targeting the nexus between driver mutations and metabolic reprogramming in cancer stem cells.

scholarly journals 2D and 3D in vitro assays to quantify the invasive behavior of glioblastoma stem cells in response to SDF-1α

BioTechniques ◽  
2020 ◽  
Vol 69 (5) ◽  
pp. 339-346
Author(s):  
Vashendriya VV Hira ◽  
Barbara Breznik ◽  
Cornelis JF Van Noorden ◽  
Tamara Lah ◽  
Remco J Molenaar
Keyword(s):  
Stem Cells ◽  
In Vitro Assays ◽  
Invasion Assay ◽  
Glioblastoma Stem Cells ◽  
Invasive Behavior ◽  
Transwell Invasion Assay ◽  
2D And 3D ◽  
Invasion Assays

Invasion is a hallmark of cancer and therefore in vitro invasion assays are important tools in cancer research. We aimed to describe in vitro 2D transwell assays and 3D spheroid assays to quantitatively determine the invasive behavior of glioblastoma stem cells in response to the chemoattractant SDF-1α. Matrigel was used as a matrix in both assays. We demonstrated quantitatively that SDF-1α increased invasive behavior of glioblastoma stem cells in both assays. We conclude that the 2D transwell invasion assay is easy to perform, fast and less complex whereas the more time-consuming 3D spheroid invasion assay is physiologically closer to the in vivo situation.

scholarly journals Circumventing drug treatment? Polyethylenimine functionalized nanoparticles inherently and selectively kill patient-derived glioblastoma stem cells

2020 ◽  
Author(s):  
Neeraj Prabhakar ◽  
Joni Merisaari ◽  
Vadim Le Joncour ◽  
Markus Peurla ◽  
Didem Sen Karaman ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesoporous Silica Nanoparticles ◽  
Treatment Modalities ◽  
Cancer Drug ◽  
Intrinsic Resistance ◽  
Functionalized Nanoparticles ◽  
Glioblastoma Stem Cells ◽  
Anti Cancer

Glioblastoma (GB) is the most frequent malignant tumor originating from the central nervous system. Despite breakthroughs in treatment modalities for other cancer types, GB remains largely irremediable due to its high degree of intratumoral heterogeneity, infiltrative growth, and intrinsic resistance towards multiple treatments. These resistant and aggressive sub-populations of GBs including the glioblastoma stem cells (GSCs) can circumvent treatment. GSCs act as a reservoir of cancer-initiating cells; they are a major challenge for successful therapy. We have discovered, as opposed to well-reported anti-cancer drug based therapeutical approach for GB therapy, the role of polyethylenimine (PEI) in inducing selective death of patient-derived GSCs via lysosomal membrane rupturing. Even at very low doses (1 ug/ml), PEI surface-functionalized mesoporous silica nanoparticles (PEI-MSNs), without any additional anti-cancer drug, very potently and selectively killed multiple GSC lines. Very importantly, PEI-MSNs did not affect the survival of well-established GB cells, or other type of cancer cells even at 25x higher doses. Remarkably, any sign of predominant cell death pathways such as apoptosis and autophagy was absent. Instead, as a potential explanation for their GSC selective killing function, we demonstrate that the internalized PEI-MSNs accumulated inside the lysosomes, subsequently causing a rupture of the vulnerable lysosomal membranes, exclusively in GSCs. As a further evaluation, we observed blood-brain-barrier (BBB) permeability of these PEI-MSNs in vitro and in vivo. Taking together the recent indications for the vulnerability of GSCs for lysosomal targeting, and GSC selectivity of the PEI-MSNs described here, the results suggest that PEI-functionalized nanoparticles could have a potential role in the eradication of GSCs.

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