Proteasome inhibitors sensitize glioma cells and glioma stem cells to TRAIL-induced apoptosis by PKCε-dependent downregulation of AKT and XIAP expressions

Abstract BackgroundGlioma stem cells (GSCs) are glioma cells with stemness and are responsible for a variety of malignant behaviors of glioma. Evidence has shown that signals from tumor microenvironment (TME) enhance stemness of glioma cells, but the identity of the signaling molecules and underlying mechanisms have been incompletely elucidated.MethodsHuman samples and glioma cell lines were cultured in vitro to determine the effects of viral infection by sphere formation, qRT-PCR, Western blot, FACS and immunofluorescence; for in vivo analysis, mice subcutaneous tumor model was carried; while bioinformatics analysis and qRT-PCR were applied for further mechanistic studies.ResultsIn this study, we show that infection of patient-derived glioma cells with adenovirus (ADV) increases the formation of tumor spheres. ADV infection upregulated stem cell markers, and the resultant tumor spheres held the capacities of self-renewal and multi-lineage differentiation, and had stronger potential to form xenograft tumors in immune-compromised mice. ADV promoted GSC formation likely via TLR9, because TLR9 was upregulated after ADV infection, and knockdown of TLR9 reduced ADV-induced GSCs. Consistently, MYD88, as well as total STAT3 and phosphorylated (p-)STAT3, were also upregulated in ADV-induced GSCs. Knockdown of MYD88 or pharmaceutical inhibition of STAT3 attenuated stemness of ADV-induced GSCs. Moreover, we found that ADV infection upregulated lncRNA NEAT1, which is downstream to TLRs and play important roles in cancer stem cells via multiple mechanisms including strengthening STAT3 signaling. Indeed, knockdown of NEAT1 impaired stemness of ADV-induced GSCs. Lastly, we show that HMGB1, a damage associated molecular pattern (DAMP) that also triggers TLR signaling, upregulated stemness markers in glioma cells.ConclusionsIn summary, our data indicate that ADV, which has been developed as vectors for gene therapy and oncolytic virus, promotes the formation of GSCs via TLR9/NEAT1/STAT3 signaling.

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DR-02 * TEMOZOLOMIDE SENSITIZES GLIOMA STEM CELLS TO TRAIL-INDUCED APOPTOSIS THROUGH UPREGULATION OF c-CBL

Neuro-Oncology ◽

10.1093/neuonc/nou252.2 ◽

2014 ◽

Vol 16 (suppl 5) ◽

pp. v63-v63

Author(s):

Z. Jing ◽

L. Li ◽

J. Liu ◽

M. Wang ◽

Y. Ban ◽

...

Keyword(s):

Stem Cells ◽

Glioma Stem Cells ◽

Induced Apoptosis

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Glioma stem cells and their non-stem differentiated glioma cells exhibit differences in mitochondrial structure and function

Oncology Reports ◽

10.3892/or.2017.6075 ◽

2017 ◽

Cited By ~ 2

Author(s):

Eun-Jung Kim ◽

Xiong Jin ◽

Ock Kim ◽

Seok Ham ◽

Sung-Hye Park ◽

...

Keyword(s):

Stem Cells ◽

Glioma Cells ◽

Structure And Function ◽

Glioma Stem Cells ◽

Mitochondrial Structure ◽

And Function

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P13.11 Transcriptional CDK inhibitors, CYC065 and THZ1 promote apoptosis in preclinical models of primary and recurrent GBM tumour cells and glioma stem cells

Neuro-Oncology ◽

10.1093/neuonc/noab180.119 ◽

2021 ◽

Vol 23 (Supplement_2) ◽

pp. ii34-ii35

Author(s):

V Juric ◽

H Duessmann ◽

H Jahns ◽

M Verreault ◽

A Idbaih ◽

...

Keyword(s):

Stem Cells ◽

Chick Embryo ◽

Cortical Neurons ◽

Tumour Cells ◽

Glioma Stem Cells ◽

Cdk Inhibitors ◽

Dependent Manner ◽

Xenograft Models ◽

Induced Apoptosis ◽

Recurrent Gbm

Abstract BACKGROUND Activation of cyclin-dependent kinases (CDKs) contributes to the uncontrolled proliferation of tumour cells. Genomic alterations that lead to the constitutive activation of CDKs are a feature of many tumours including glioblastoma (GBM), the most common and aggressive primary brain tumour. Patient resistance to the current standard of care, temozolomide and radiotherapy, is common and highlights the need to discover more effective treatment strategies. Additionally, glioma stem cells (GSCs), tumour initiating cells contribute to treatment resistance in GBM. Targeting GBM tumour cells and GSCs using transcriptional CDK inhibitors, CYC065 and THZ1 is a potential novel treatment to prevent relapse of the tumour. MATERIAL AND METHODS The therapeutic efficacy of two CDK inhibitors (CKIs) was tested in a panel of ten low-passage GBM patient-derived gliomasphere cultures and semi-in vivo chick embryo xenograft models. Specifically, transcriptional inhibitors targeting CDK9/2 (CYC065) and CDK7 (THZ1) were used. Mechanism of cell death was examined following CKI treatment. Additionally, dependence on anti-apoptotic proteins was studied using genetic depletion and BH3 profiling. Fluorescence activated cell sorting (FACS) and light-sheet fluorescence microscopy (LSFM) were employed to study stem cell populations in recurrent GBM and evaluation of CKI efficacy against GSCs. RESULTS We here demonstrate that CYC065 and THZ1 treatments cause loss of cell viability and induce caspase-dependent apoptosis in primary and recurrent patient-derived gliomaspheres while sparing primary cortical neurons. Importantly, apoptosis responses manifested across a range of time points that significantly correlated with the cell doubling time. Mechanistically, CYC065 and THZ1 downregulate the anti-apoptotic protein Mcl-1, which suffices to sensitise gliomasphere cultures to treatment-induced apoptosis in a Bim-dependent manner. Additionally, high expression levels of chemo- and radio-resistant GSCs were found in recurrent gliomaspheres. CKIs induced apoptosis in isolated CD133 and CD44 biomarker-positive cells while TMZ was ineffective, highlighting the potential of these drugs to overcome resistance to conventional chemotherapy. Additionally, using LSFM we shown that CD133, CD44 GSC biomarker-negative cells convert into GSC biomarker-positive cells and contribute to the enrichment in GSCs in recurrent GBM which could potentially explain the TMZ ineffectiveness. Finally, CKIs reduced proliferation and promoted apoptosis in chick embryo xenograft models of primary and recurrent GBM. CONCLUSION Collectively, these data demonstrate that CYC065 and THZ1 display high anti-cancer activity in primary and recurrent GBM and provide scientific rationale for the further development of CDK inhibitors to potentiate their clinical utilization in the future.

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The Roles of miRNA in Glioblastoma Tumor Cell Communication: Diplomatic and Aggressive Negotiations

International Journal of Molecular Sciences ◽

10.3390/ijms21061950 ◽

2020 ◽

Vol 21 (6) ◽

pp. 1950 ◽

Cited By ~ 4

Author(s):

Andrei Buruiană ◽

Ștefan Ioan Florian ◽

Alexandru Ioan Florian ◽

Teodora-Larisa Timiș ◽

Carmen Mihaela Mihu ◽

...

Keyword(s):

Stem Cells ◽

Endothelial Cells ◽

Cell Communication ◽

Glioma Cells ◽

Glioma Stem Cells ◽

Patient Stratification ◽

Egfr Signaling ◽

Egfr Signaling Pathway ◽

Target Therapies ◽

Novel Target

Glioblastoma (GBM) consists of a heterogeneous collection of competing cellular clones which communicate with each other and with the tumor microenvironment (TME). MicroRNAs (miRNAs) present various exchange mechanisms: free miRNA, extracellular vesicles (EVs), or gap junctions (GJs). GBM cells transfer miR-4519 and miR-5096 to astrocytes through GJs. Oligodendrocytes located in the invasion front present high levels of miR-219-5p, miR-219-2-3p, and miR-338-3p, all related to their differentiation. There is a reciprocal exchange between GBM cells and endothelial cells (ECs) as miR-5096 promotes angiogenesis after being transferred into ECs, whereas miR-145-5p acts as a tumor suppressor. In glioma stem cells (GSCs), miR-1587 and miR-3620-5p increase the proliferation and miR-1587 inhibits the hormone receptor co-repressor-1 (NCOR1) after EVs transfers. GBM-derived EVs carry miR-21 and miR-451 that are up-taken by microglia and monocytes/macrophages, promoting their proliferation. Macrophages release EVs enriched in miR-21 that are transferred to glioma cells. This bidirectional miR-21 exchange increases STAT3 activity in GBM cells and macrophages, promoting invasion, proliferation, angiogenesis, and resistance to treatment. miR-1238 is upregulated in resistant GBM clones and their EVs, conferring resistance to adjacent cells via the CAV1/EGFR signaling pathway. Decrypting these mechanisms could lead to a better patient stratification and the development of novel target therapies.

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Topoisomerase I Inhibitors, Shikonin and Topotecan, Inhibit Growth and Induce Apoptosis of Glioma Cells and Glioma Stem Cells

PLoS ONE ◽

10.1371/journal.pone.0081815 ◽

2013 ◽

Vol 8 (11) ◽

pp. e81815 ◽

Cited By ~ 30

Author(s):

Feng-Lei Zhang ◽

Ping Wang ◽

Yun-Hui Liu ◽

Li-bo Liu ◽

Xiao-Bai Liu ◽

...

Keyword(s):

Stem Cells ◽

Topoisomerase I ◽

Glioma Cells ◽

Glioma Stem Cells ◽

Topoisomerase I Inhibitors

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Abstract 4241: Glioma stem cells perform oxidative phosphorylation, while the rest of glioma cells perform aerobic glycolysis

10.1158/1538-7445.am10-4241 ◽

2010 ◽

Author(s):

Erina Vlashi ◽

Chann Lagadec ◽

Lorenza Della Donna ◽

YongHong Meng ◽

Carmen Dekmezian ◽

...

Keyword(s):

Stem Cells ◽

Oxidative Phosphorylation ◽

Aerobic Glycolysis ◽

Glioma Cells ◽

Glioma Stem Cells

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Role of mesenchymal stem cells in delivering Newcastle disease virus to glioma cells and glioma stem cells and enhancing the oncolytic effect of the virus by secreting TRAIL.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.15_suppl.3100 ◽

2013 ◽

Vol 31 (15_suppl) ◽

pp. 3100-3100

Author(s):

Shimon Slavin ◽

Gila Kazimirsky ◽

Amotz Ziv-Av ◽

Chaya Brodie

Keyword(s):

Stem Cells ◽

Cell Death ◽

Newcastle Disease Virus ◽

Cancer Cells ◽

Disease Virus ◽

Newcastle Disease ◽

Glioma Cells ◽

Antibody Array ◽

Glioma Stem Cells ◽

Oncolytic Effect

3100 Background: Newcastle disease virus (NDV), an avian paramyxovirus, is tumor selective and oncolytic by induction of apoptosis. Preclinical and clinical studies in patients with glioblastoma (GBM) using NDV demonstrated occasional clinical benefits with no major side effects. Limitations to the use of NDV as virotherapy of GBM is the inefficient delivery into cancer cells in the brain. Methods: Mesenchymal stromal cells (MSCs) can migrate towards cancer cells. We examined potential delivery of oncolytic effect of NDV (MTH-68/H) against glioma cell lines and glioma stem cells (GSCs) and the ability of MSCs to deliver NDV to glioma cells and GSCs in culture. Results: NDV induced a dose-dependent cell death in the glioma cells U87, A172 and U251 with maximal effects at 10 MOI. In contrast, we found only small level of apoptosis or changes in self-renewal in three GSCs infected with NDV. We found that MSCs derived from bone marrow, adipose tissue and cord were successfully infected by NDV and were able to deliver the virus to co-cultured glioma cells and GSCs. In addition, treatment of glioma cells and GSCs with culture supernatant of infected MSCs increase apoptosis of glioma cells as compared to the effect of direct infection of glioma cells. Moreover, the culture supernatants of the infected MSCs induced cell death in GSCs that were resistant to the oncolytic effect of NDV, suggesting that factor(s) secreted by the infected MSCs sensitized the glioma cells and GSCs to the cytotoxic effects of NDV. Using antibody array and ELISA we identified TRAIL as the factor secreted from infected MSCs. Indeed, treatment of infected glioma cells with TRAIL increased the cytotoxic effect of NDV and sensitized GSCs to the oncolytic effects of NDV. Conclusions: MSCs can be employed to deliver NDV to GBM. In addition, MSCs can also sensitize glioma cells and GSCs to oncolysis by NDV. Considering the resistance of GSCs to chemotherapy and radiation therapy, treatment of GBM with MSC-mediated targeted oncolytic NDV may provide a new clinical tool for treatment of GBM and eradication of GSCs.

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