Effects of Chemotherapeutics on Organotypic Corticostriatal Slice Cultures Identified by A Panel of Fluorescent and Immunohistochemical Markers

Nørregaard, Annette; Jensen, Stine Skov; Kolenda, Jesper; Aaberg-Jessen, Charlotte; Christensen, Knud Villy; Jensen, Poul Henning; Kristensen, Bjarne Winther

doi:10.1007/s12640-011-9300-9

Cited by 12 publications

(11 citation statements)

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“…Organotypic corticostriatal slice cultures were prepared as previous described [13, 14]. Seven days after the start of culturing the brain slice cultures were exposed to siramesine and cell death was examined by PI uptake as described earlier by Nørregaard et al [14].…”

Section: Methodsmentioning

confidence: 99%

“…Seven days after the start of culturing the brain slice cultures were exposed to siramesine and cell death was examined by PI uptake as described earlier by Nørregaard et al [14]. …”

Section: Methodsmentioning

confidence: 99%

“…Accordingly, we used both standard glioma cell lines and patient-derived spheroids cultures with tumor-initiating stem-like cells [12, 13]. To perform a thorough testing, spheroid cultures were implanted in three dimensionel organotypic brain slice cultures [13, 14] and used for generation of patient-like tumors in a glioblastoma xenograft mice model [12, 15, 16]. Using these approaches the in vitro results with tumor cells and spheroids suggested a potential of lysosomal destabilizing drugs in killing glioblastoma cells, but in the organotypic spheroid-brain slice culture model and the in vivo xenograft model siramesine was without effect.…”

Section: Introductionmentioning

confidence: 99%

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Effects of the lysosomal destabilizing drug siramesine on glioblastoma in vitro and in vivo

et al. 2017

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BackgroundGlioblastoma is the most frequent and most malignant brain tumor with the patients having a median survival of only 14.6 months. Although glioblastoma patients are treated with surgery, radiation and chemotherapy recurrence is inevitable. A stem-like population of radio- and chemoresistant brain tumor-initiating cells combined with the invasive properties of the tumors is believed to be critical for treatment resistance. In the present study, the aim was to investigate the effect of a novel therapeutic strategy using the lysosomotropic detergent siramesine on glioblastomas.MethodsStandard glioma cell lines and patient-derived spheroids cultures with tumor-initiating stem-like cells were used to investigate effects of siramesine on proliferation and cell death. Responsible mechanisms were investigated by inhibitors of caspases and cathepsins. Effects of siramesine on migrating tumor cells were investigated by a flat surface migration assay and by implanting spheroids into organotypic rat brain slice cultures followed by confocal time-lapse imaging. Finally the effect of siramesine was investigated in an orthotopic mouse glioblastoma model. Results obtained in vitro and in vivo were confirmed by immunohistochemical staining of histological sections of spheroids, spheroids in brain slice cultures and tumors in mice brains.ResultsThe results showed that siramesine killed standard glioma cell lines in vitro, and loss of acridine orange staining suggested a compromised lysosomal membrane. Co-treatment of the cell lines with inhibitors of caspases and cathepsins suggested differential involvement in cell death. Siramesine caused tumor cell death and reduced secondary spheroid formation of patient-derived spheroid cultures. In the flat surface migration model siramesine caused tumor cell death and inhibited tumor cell migration. This could not be reproduced in the organotypic three dimensional spheroid-brain slice culture model or in the mice xenograft model.ConclusionsIn conclusion the in vitro results obtained with tumor cells and spheroids suggest a potential of lysosomal destabilizing drugs in killing glioblastoma cells, but siramesine was without effect in the organotypic spheroid-brain slice culture model and the in vivo xenograft model.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-017-3162-3) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

“…Seven days after the start of culturing the brain slice cultures were exposed to siramesine and cell death was examined by PI uptake as described earlier by Nørregaard et al [14]. …”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effects of the lysosomal destabilizing drug siramesine on glioblastoma in vitro and in vivo

et al. 2017

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“…Noteworthy, many previous studies ensure neurons safety by evaluating single morphological markers. 37 , 38 , 39 , 40 In some studies the amino-acid profile of treated cells has been quantified. 41 , 42 In this report, we applied the synaptic optogenetic function analysis (SOFA) tool to unravel sub-morphological changes of neurons.…”

Section: Introductionmentioning

confidence: 99%

Chemotherapeutic xCT inhibitors sorafenib and erastin unraveled with the synaptic optogenetic function analysis tool

et al. 2017

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In the search for new potential chemotherapeutics, the compounds’ toxicity to healthy cells is an important factor. The brain with its functional units, the neurons, is especially endangered during the radio- and chemotherapeutic treatment of brain tumors. The effect of the potential compounds not only on neuronal survival but also neuronal function needs to be taken into account. Therefore, in this study we aimed to comprehend the biological effects of chemotherapeutic xCT inhibition on healthy neuronal cells with our synaptic optogenetic function analysis tool (SOFA). We combined common approaches, such as investigation of morphological markers, neuronal function and cell metabolism. The glutamate-cystine exchanger xCT (SLC7A11, system Xc−) is the main glutamate exporter in malignant brain tumors and as such a relevant drug target for treating deadly glioblastomas (WHO grades III and IV). Recently, two small molecules termed sorafenib (Nexavar) and erastin have been found to efficiently block xCT function. We investigated neuronal morphology, metabolic secretome profiles, synaptic function and cell metabolism of primary hippocampal cultures (containing neurons and glial cells) treated with sorafenib and erastin in clinically relevant concentrations. We found that sorafenib severely damaged neurons already after 24 h of treatment. Noteworthy, also at a lower concentration, where no morphological damage or metabolic disturbance was monitored, sorafenib still interfered with synaptic and metabolic homeostasis. In contrast, erastin-treated neurons displayed mostly inconspicuous morphology and metabolic rates. Key parameters of proper neuronal function, such as synaptic vesicle pool sizes, were however disrupted following erastin application. In conclusion, our data revealed that while sorafenib and erastin effectively inhibited xCT function they also interfered with essential neuronal (synaptic) function. These findings highlight the particular importance of investigating the effects of potential neurooncological and general cancer chemotherapeutics also on healthy neuronal cells and their function as revealed by the SOFA tool.

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“…3, 5-tetrazin-4(3H) -one (temozolomide, TMZ) because of its high permeability across blood-brain barrier (BBB) and good cytotoxic activity for gliomas [12]- [13]. BCNU always induces two major types of genotoxic damage to the cell: DNA mono adducts and DNA interstrand cross-links.…”

Section: Introductionmentioning

confidence: 99%

Nimustine Induces DNA Breaks and Crosslinks in NIH/3T3 Cells

Zhao¹,

Chen²,

Zhong³

et al. 2013

IJBBB

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Effects of Chemotherapeutics on Organotypic Corticostriatal Slice Cultures Identified by A Panel of Fluorescent and Immunohistochemical Markers

Cited by 12 publications

References 78 publications

Effects of the lysosomal destabilizing drug siramesine on glioblastoma in vitro and in vivo

Effects of the lysosomal destabilizing drug siramesine on glioblastoma in vitro and in vivo

Chemotherapeutic xCT inhibitors sorafenib and erastin unraveled with the synaptic optogenetic function analysis tool

Nimustine Induces DNA Breaks and Crosslinks in NIH/3T3 Cells

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