Transcriptomic Profiling of DNA Damage Response in Patient-Derived Glioblastoma Cells before and after Radiation and Temozolomide Treatment

Lozinski, Mathew; Graves, Moira C.; Fay, Michael; Day, Bryan W.; Stringer, Brett W.; Tooney, Paul A.

doi:10.3390/cells11071215

Cited by 9 publications

(10 citation statements)

References 42 publications

(56 reference statements)

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“…This lack of synergistic interaction observed in TMZ+RT treated cell lines is worth noting and has been observed in U251 and U373MG cell lines [ 41 , 42 ]. Our previous work showed that glioblastoma cells treated with TMZ or RT elicits different patterns of DDR upregulation, and when combined, a lower response of DDR genes is observed [ 43 ]. Thus, the addition of DNA-damaging agents (i.e., TMZ and RT) that induce slightly different but overlapping activation of DDR pathways at different time points may hinder synergistic responses, as the activation of DDR from one agent may prime the cell for repair of the damage induced by the other.…”

Section: Discussionmentioning

confidence: 99%

ATR inhibition using gartisertib enhances cell death and synergises with temozolomide and radiation in patient-derived glioblastoma cell lines

Lozinski,

Bowden,

Graves

et al. 2024

Oncotarget

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Glioblastoma cells can restrict the DNA-damaging effects of temozolomide (TMZ) and radiation therapy (RT) using the DNA damage response (DDR) mechanism which activates cell cycle arrest and DNA repair pathways. Ataxia-telangiectasia and Rad3-Related protein (ATR) plays a pivotal role in the recognition of DNA damage induced by chemotherapy and radiation causing downstream DDR activation. Here, we investigated the activity of gartisertib, a potent ATR inhibitor, alone and in combination with TMZ and/or RT in 12 patient-derived glioblastoma cell lines. We showed that gartisertib alone potently reduced the cell viability of glioblastoma cell lines, where sensitivity was associated with the frequency of DDR mutations and higher expression of the G2 cell cycle pathway. ATR inhibition significantly enhanced cell death in combination with TMZ and RT and was shown to have higher synergy than TMZ+RT treatment. MGMT promoter unmethylated and TMZ+RT resistant glioblastoma cells were also more sensitive to gartisertib. Analysis of gene expression from gartisertib treated glioblastoma cells identified the upregulation of innate immune-related pathways. Overall, this study identifies ATR inhibition as a strategy to enhance the DNA-damaging ability of glioblastoma standard treatment, while providing preliminary evidence that ATR inhibition induces an innate immune gene signature that warrants further investigation.

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Section: Discussionmentioning

confidence: 99%

ATR inhibition using gartisertib enhances cell death and synergises with temozolomide and radiation in patient-derived glioblastoma cell lines

Lozinski,

Bowden,

Graves

et al. 2024

Oncotarget

Self Cite

View full text Add to dashboard Cite

show abstract

“…This lack of synergistic interaction observed in TMZ+RT treated cell lines is worth noting, and has been observed in U251 and U373MG cell lines 43,44 . Our previous work showed that glioblastoma cells treated with TMZ or RT elicits different patterns of DDR upregulation, and when combined, a lower response of DDR genes is observed 21 . Thus, the addition of DNA-damaging agents (i.e., TMZ and RT) that induce slightly different but overlapping activation of DDR pathways at different time points may hinder synergistic responses, as the activation of DDR from one agent may prime the cell for repair of the damage induced by the other.…”

Section: Discussionmentioning

confidence: 99%

“…The 7-day time point was chosen as untreated cells reached a high level of confluence after cell seeding by day 7. Cell viability was assessed using the MTT cell viability assay as previously described 21 . The IC50 of gartisertib and berzosertib in each respective cell line was determined using the MTT cell viability assay.…”

Section: Cell Viability Assaymentioning

confidence: 99%

ATR inhibition using gartisertib enhances cell death and synergises with temozolomide and radiation in patient-derived glioblastoma cell lines

Lozinski,

Bowden,

Graves

et al. 2023

Preprint

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Glioblastoma cells can restrict the DNA-damaging effects of temozolomide (TMZ) and radiation therapy (RT) using the DNA damage response (DDR) mechanism which activates cell cycle arrest and DNA repair pathways. Ataxia-telangiectasia and Rad3-Related protein (ATR) plays a pivotal role in the recognition of DNA damage induced by chemotherapy and radiation causing downstream DDR activation. Here, we investigated the activity of gartisertib, a potent ATR inhibitor, alone and in combination with TMZ and/or RT in 12 patient-derived glioblastoma cell lines. We showed that gartisertib alone potently reduced the cell viability of glioblastoma cell lines, where sensitivity was associated with the frequency of DDR mutations and higher expression of the G2 cell cycle pathway. ATR inhibition significantly enhanced cell death in combination with TMZ and RT and was shown to have higher synergy than TMZ+RT treatment.MGMTpromoter unmethylated and TMZ+RT resistant glioblastoma cells were also more sensitive to gartisertib. Analysis of gene expression from gartisertib treated glioblastoma cells identified the upregulation of innate immune-related pathways. Overall, this study identifies ATR inhibition as a strategy to enhance the DNA-damaging ability of glioblastoma standard treatment, while providing preliminary evidence that ATR inhibition induces an innate immune gene signature that warrants further investigation.

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“…7) RPA3 is part of the three-subunit replication protein A (RPA) complex involved in homologous recombination and double strand break repair, and has been implicated in glioblastoma survival outcomes. 40 FoxM1 has been reported to promote stemness, radioresistance and mesenchymal transition in GBM. Radioresistance in GBM is known to be conferred by glioblastoma stem-like cells and is a key factor of GBM treatment resistance.…”

Section: Effect Of Rpa3 and Blvra Silencing In Npcsmentioning

confidence: 99%

A Multistep In Silico Approach Identifies Potential Glioblastoma Drug Candidates via Inclusive Molecular Targeting of Glioblastoma Stem Cells

Dogra,

Singh,

Kumar

2024

Mol Neurobiol

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Aims Glioblastoma (GBM) is the highest-grade of glioma for which no effective therapy is currently available. Despite extensive research in diagnosis and therapy there has been no signi cant improvement in GBM outcomes, with a median overall survival continuing at a dismal 15-18 months. In recent times, glioblastoma stem cells (GSCs) have been identi ed as crucial drivers of treatment resistance and tumor recurrence; and GBM therapies targeting GSCs are expected to improve patient outcomes. We used a multistep screening strategy to identify repurposed candidate drugs against therapeutic molecular targets in GBM with potential to concomitantly target GSCs. Main methods Common differentially expressed genes (DEGs) were identi ed through analysis of multiple GBM and GSC datasets from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA). For identi cation of target genes, we further selected the genes with most signi cant effect on overall patient survival. The relative mRNA and protein expression of the selected genes in TCGA control versus GBM samples was also validated and their cancer dependency scores were assessed. Drugs targeting these genes and their corresponding proteins were identi ed from LINCS database using Connectivity Map (CMap) portal, and by in silico molecular docking against each individual target using FDA approved drugs library from DrugBank database, respectively. The molecules thus obtained were further evaluated for their ability to cross blood brain barrier (BBB) and their likelihood of resulting in drug resistance by acting as p-glycoprotein (p-Gp) substrates. The effect of these nal shortlisted compounds on a panel of GBM cell lines was examined and compared with temozolomide through the drug sensitivity EC50 values and AUC from the PRISM Repurposing Secondary Screen, and IC50 values obtained from GDSC portal. Key ndings We identi ed RPA3, PSMA2, PSMC2, BLVRA and HUS1 as molecular targets in GBM including GSCs with signi cant impact on patient survival. Our results show GSK-2126458, linifanib, drospirenone, eltrombopag, nilotinib and PD198306 as candidate drugs which can be further evaluated for their anti-tumor potential against GBM. Signi cance Through this work we identi ed repurposed candidate therapeutics against GBM utilizing a GSC inclusive targeting approach, which demonstrated high in vitro e cacy. These drugs have the potential to be developed as individual or combination therapy to improve GBM outcomes.

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Transcriptomic Profiling of DNA Damage Response in Patient-Derived Glioblastoma Cells before and after Radiation and Temozolomide Treatment

Cited by 9 publications

References 42 publications

ATR inhibition using gartisertib enhances cell death and synergises with temozolomide and radiation in patient-derived glioblastoma cell lines

ATR inhibition using gartisertib enhances cell death and synergises with temozolomide and radiation in patient-derived glioblastoma cell lines

ATR inhibition using gartisertib enhances cell death and synergises with temozolomide and radiation in patient-derived glioblastoma cell lines

A Multistep In Silico Approach Identifies Potential Glioblastoma Drug Candidates via Inclusive Molecular Targeting of Glioblastoma Stem Cells

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