Phospho-mTOR expression in human glioblastoma microglia-macrophage cells

Lisi, Lucia; Ciotti, Gabriella Maria Pia; Chiavari, Marta; Pizzoferrato, Michela; Mangiola, Annunziato; Kalinin, Sergey; Feinstein, Douglas L.; Navarra, Pierluigi

doi:10.1016/j.neuint.2019.104485

Cited by 20 publications

(23 citation statements)

References 34 publications

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“…In the last years, our group has investigated the role of GAMs in murine and human models of GB microenvironment, looking at different regulatory molecules involved in GAM polarization and activation status [26][27][28][29][30]. In the present study, we have evaluated both in GB cells and microglia the expression of PDIA3 in GB specimens collected from patients after surgical resection of the tumor.…”

Section: Introductionmentioning

confidence: 98%

PDIA3 Expression in Glioblastoma Modulates Macrophage/Microglia Pro-Tumor Activation

Chiavari¹,

Ciotti²,

Canonico

et al. 2020

IJMS

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The glioblastoma (GB) microenvironment includes cells of the innate immune system identified as glioma-associated microglia/macrophages (GAMs) that are still poorly characterized. A potential role on the mechanisms regulating GAM activity might be played by the endoplasmic reticulum protein ERp57/PDIA3 (protein disulfide-isomerase A3), the modulation of which has been reported in a variety of cancers. Moreover, by using The Cancer Genome Atlas database, we found that overexpression of PDIA3 correlated with about 55% reduction of overall survival of glioma patients. Therefore, we analyzed the expression of ERp57/PDIA3 using specimens obtained after surgery from 18 GB patients. Immunohistochemical analysis of tumor samples revealed ERp57/PDIA3 expression in GB cells as well as in GAMs. The ERp57/PDIA3 levels were higher in GAMs than in the microglia present in the surrounding parenchyma. Therefore, we studied the role of PDIA3 modulation in microglia–glioma interaction, based on the ability of conditioned media collected from human GB cells to induce the activation of microglial cells. The results indicated that reduced PDIA3 expression/activity in GB cells significantly limited the microglia pro-tumor polarization towards the M2 phenotype and the production of pro-inflammatory factors. Our data support a role of PDIA3 expression in GB-mediated protumor activation of microglia.

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Section: Introductionmentioning

confidence: 98%

PDIA3 Expression in Glioblastoma Modulates Macrophage/Microglia Pro-Tumor Activation

Chiavari¹,

Ciotti²,

Canonico

et al. 2020

IJMS

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“…mTOR signalling is known to regulate the balance between pro‐ and anti‐inflammatory responses and may be responsible for the dysregulated inflammatory response in TAM, which display a shift towards anti‐inflammatory activity. Interestingly, increased mTOR phosphorylation at Ser‐2448 is present in nearly 40% of TAM in human GBM (Lisi et al , ); however, the functional impact of this mTOR deregulation and its molecular mechanism have never been characterised.…”

Section: Introductionmentioning

confidence: 99%

Microglia promote glioblastoma via mTOR‐mediated immunosuppression of the tumour microenvironment

et al. 2020

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Tumour‐associated microglia/macrophages (TAM) are the most numerous non‐neoplastic populations in the tumour microenvironment in glioblastoma multiforme (GBM), the most common malignant brain tumour in adulthood. The mTOR pathway, an important regulator of cell survival/proliferation, is upregulated in GBM, but little is known about the potential role of this pathway in TAM. Here, we show that GBM‐initiating cells induce mTOR signalling in the microglia but not bone marrow‐derived macrophages in both in vitro and in vivo GBM mouse models. mTOR‐dependent regulation of STAT3 and NF‐κB activity promotes an immunosuppressive microglial phenotype. This hinders effector T‐cell infiltration, proliferation and immune reactivity, thereby contributing to tumour immune evasion and promoting tumour growth in mouse models. The translational value of our results is demonstrated in whole transcriptome datasets of human GBM and in a novel in vitro model, whereby expanded‐potential stem cells (EPSC)‐derived microglia‐like cells are conditioned by syngeneic patient‐derived GBM‐initiating cells. These results raise the possibility that microglia could be the primary target of mTOR inhibition, rather than the intrinsic tumour cells in GBM.

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“…Arginase-1 (ARG1) converts arginine to ornithine and urea, competing with nitric oxide synthase (NOS), usually used as M1 marker which utilizes arginine to produce nitric oxide [43; 44]. Our previous data shown an increase of urea production when CHME-5 were treated with CMs from wild type T98G, reinforcing the M2 phenotype [28]. Here, we interestingly reveal a phenotypic modi cation of CHME-5 by detaching the cells from the glioma-induced M2 phenotype with changing in urea production and in morphology.…”

Section: Discussionmentioning

confidence: 99%

“…All the experiments received institutional approval. Conditioned media from activated glioma cells was generated following a protocol described before [28]. Brie y, Basal Conditioned Media (B-CM) was prepared with 4 hours incubation in plain medium, followed by 3 washes with phosphate buffered saline (PBS) and addition of fresh plain medium for 24 hours.…”

Section: Cell Culturesmentioning

confidence: 99%

PDIA3 Reduce Glioma-associated Macrophage/microglia Pro-tumor Activation trough IL-6-STAT3-PDIA3 Pathway.

Chiavari

Ciotti

Canonico

et al. 2020

Preprint

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Background: Glioblastoma (GB - grade IV glioma) is the most aggressive and common cancer of central nervous system with an overall survival of 14-16 months. The GB tumor microenvironment includes cells of the innate immune system identified as glioma-associated microglia/macrophages (GAMs). It is known that between GAMs and GB cells there is a double interaction, but the role of GAMs is still poorly characterized. The endoplasmic reticulum (ER) protein ERp57, also known as PDIA3, is a thiol oxidoreductase with main function related on glycoprotein folding in endoplasmic reticulum. However, PDIA3 shows different functions. In fact, the various subcellular localizations and binding partners of PDIA3 affect numerous physiological processes and diseases: different regulation and modulation of PDIA3 has been reported in multiple pathologies including neurodegenerative diseases and cancer. Methods: In the present work, we evaluated in both GB cells and microglia-macrophage cells the expression of PDIA3 using specimens collected after surgical from 18 GB patients. In addition, we studied in vitro microglia-glioma interaction to determine the role of PDIA3 in viability and the activation of both GB and microglia cells. The study was carried using PDIA3-silenced T98G cells and/or using a pharmacological inhibitor of PDIA3 activity (Punicalagin-PUN).Results: We initially investigated the role of the PDIA3 in GB survival by inquiring The Cancer Genome Atlas dataset. The results indicated that 352 out of 690 patients reported over-expression of PDIA3, which significantly correlated with a ~55% reduction of overall survival. Subsequently, for the first time, we investigated the PDIA3 expression in the tumor and the nearby parenchyma of 18 GB patients and our data showed a significant upregulation (15% vs 10%) of ERp57/PDIA3 in GAMs of tumor specimens respect the microglia present in parenchyma. In addition, we show that conditioned medium (CMs) obtained from both wild type T98G and PDIA3 silenced T98G induced an activation of microglia cells, but the PDIA3 silenced-T98G CMs significant limited the microglia pro-tumor activation probably through a IL-6-STAT3-PDIA3 dependent mechanism. Conclusion: Our data support the relevant role of PDIA3 expression in GB pathology and link the different activation of microglia to a mechanism a IL-6-STAT3-PDIA3 dependent.

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Phospho-mTOR expression in human glioblastoma microglia-macrophage cells

Cited by 20 publications

References 34 publications

PDIA3 Expression in Glioblastoma Modulates Macrophage/Microglia Pro-Tumor Activation

PDIA3 Expression in Glioblastoma Modulates Macrophage/Microglia Pro-Tumor Activation

Microglia promote glioblastoma via mTOR‐mediated immunosuppression of the tumour microenvironment

PDIA3 Reduce Glioma-associated Macrophage/microglia Pro-tumor Activation trough IL-6-STAT3-PDIA3 Pathway.

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