P13.01 Neuronal activity drives distinct invasion modes of glioma cells

Abstract BACKGROUND Gliomas are incurable brain tumors characterized by their infiltrative growth which makes them a whole-brain disease. Previously we described membrane protrusions called tumor microtubes (TMs), and glutamatergic synapses between neurons and glioma cells, as mechanisms contributing to glioma cell invasion and tumor progression. However, the interrelation of the two, and the exact mechanisms of glioma cell dynamics over time was unknown. Therefore, we investigate neuronal synaptic input on TM-associated glioma cell motility. MATERIAL AND METHODS Here we established a novel workflow for analyzing single glioma cell dynamics over several hours with in-vivo two-photon microscopy. First, a membranous fluorescent marking of patient-derived glioma cells was established to reliably track membrane changes. Secondly, augmented microscopy based on deep- and machine-learning algorithms was used to track glioma cells. Neuronal activity was manipulated with different doses of isoflurane anesthesia, and used to study its effects on glioma cell dynamics. RESULTS This novel method revealed that motility of glioma cells can be described by the displacement of whole glioma cell somata (somatokinesis) and TM dynamics. TM motility in turn could be sub-categorized into protrusion, retraction and branching. Next, we describe three different invasion modes, all with similarities to different cell types involved in CNS development. Lastly, the effects of neuronal activity on glioma cell invasion were investigated. With the application of high anesthesia and subsequently reduced neuronal activity, TM turnover, branching events and as a result glioma cell invasion were inhibited, but in a heterogeneous manner. CONCLUSION The novel workflow allowed to comprehensively characterize glioma cell invasion over several hours. Its application demonstrates novel, hitherto unknown cellular mechanisms of glioma cell invasion, and provides a link between TM biology and neuron-glioma communication. Finally, neuronal input drives distinct subtypes of glioma cell motility patterns.All in all, this work presents an important first step in understanding mechanisms that lead to the whole- brain colonization of glioma cells making these brain tumors incurable. A further characterization of the exact molecular mechanisms that drive neuronal activity-dependent glioma cell motility is warranted.

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TAMI-52. NEURONAL MECHANISMS OF BRAIN TUMOR INVASION

Neuro-Oncology ◽

10.1093/neuonc/noab196.835 ◽

2021 ◽

Vol 23 (Supplement_6) ◽

pp. vi209-vi209

Author(s):

Varun Venkataramani ◽

Yvonne Yang ◽

Marc Schubert ◽

Carlo Beretta ◽

Michael Botz ◽

...

Keyword(s):

Brain Tumor ◽

Neuronal Activity ◽

Cell Invasion ◽

Glioma Cell ◽

Tumour Progression ◽

Glioma Cells ◽

Movement Patterns ◽

Calcium Transients ◽

High Temporal Resolution ◽

Branching Points

Abstract Incurable gliomas are characterized by their infiltration into the whole brain. Recently, we described tumor microtubes as a novel structure contributing to glioma cell invasion and uncovered synaptic contacts on glioma cells that drive brain tumour progression. However, the exact effects of neuronal activity on glioma cell motility are yet unclear. Here, we show how a recently described neuronal-like cellular transcription state of glioblastoma cells is correlated to glioma cell invasion in vivo. To unravel the details of neuronal features of glioma invasion in space and time, we established a novel approach of intravital imaging for brain tumor cells with a membrane-bound GFP combined with deep learning algorithms that are used to track glioma cell processes with a high temporal resolution over several hours. This approach uncovers how invading tumor microtubes use Levy-like movement patterns indicative of efficient search patterns often employed by animal predators searching for scarce resources such as food. Neuronal activity is able to accelerate the tumor microtube dynamics, accelerate the Levy-like movement patterns and increase the overall invasion speed of glioma cells. These processes are mediated by local calcium transients in glioma cell somata and tumor microtubes. In accordance, genetic manipulation and pharmacological perturbation of AMPA receptors reduces tumor microtube length, number and branching points by interfering with intracellular calcium transients. All in all, the work here uncovers novel neuronal activity-mediated mechanisms of glioma cell invasion, a hallmark of this yet fatal disease.

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Pisosterol Induces G2/M Cell Cycle Arrest and Apoptosis via the ATM/ATR Signaling Pathway in Human Glioma Cells

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200203160117 ◽

2020 ◽

Vol 20 (6) ◽

pp. 734-750

Author(s):

Wallax A.S. Ferreira ◽

Rommel R. Burbano ◽

Claudia do Ó. Pessoa ◽

Maria L. Harada ◽

Bárbara do Nascimento Borges ◽

...

Keyword(s):

Cell Cycle ◽

Cell Cycle Arrest ◽

Signaling Pathway ◽

Glioma Cell ◽

Molecular Mechanisms ◽

Glioma Cells ◽

Dependent Manner ◽

M Cell ◽

Trypan Blue Exclusion ◽

Cycle Arrest

Background: Pisosterol, a triterpene derived from Pisolithus tinctorius, exhibits potential antitumor activity in various malignancies. However, the molecular mechanisms that mediate the pisosterol-specific effects on glioma cells remain unknown. Objective: This study aimed to evaluate the antitumoral effects of pisosterol on glioma cell lines. Methods: The 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) and trypan blue exclusion assays were used to evaluate the effect of pisosterol on cell proliferation and viability in glioma cells. The effect of pisosterol on the distribution of the cells in the cell cycle was performed by flow cytometry. The expression and methylation pattern of the promoter region of MYC, ATM, BCL2, BMI1, CASP3, CDK1, CDKN1A, CDKN2A, CDKN2B, CHEK1, MDM2, p14ARF and TP53 was analyzed by RT-qPCR, western blotting and bisulfite sequencing PCR (BSP-PCR). Results: Here, it has been reported that pisosterol markedly induced G2/M arrest and apoptosis and decreased the cell viability and proliferation potential of glioma cells in a dose-dependent manner by increasing the expression of ATM, CASP3, CDK1, CDKN1A, CDKN2A, CDKN2B, CHEK1, p14ARF and TP53 and decreasing the expression of MYC, BCL2, BMI1 and MDM2. Pisosterol also triggered both caspase-independent and caspase-dependent apoptotic pathways by regulating the expression of Bcl-2 and activating caspase-3 and p53. Conclusions: It has been, for the first time, confirmed that the ATM/ATR signaling pathway is a critical mechanism for G2/M arrest in pisosterol-induced glioma cell cycle arrest and suggests that this compound might be a promising anticancer candidate for further investigation.

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miR-203 sensitizes glioma cells to temozolomide and inhibits glioma cell invasion by targeting E2F3

Molecular Medicine Reports ◽

10.3892/mmr.2014.3101 ◽

2014 ◽

Vol 11 (4) ◽

pp. 2838-2844 ◽

Cited By ~ 9

Author(s):

GUODONG TANG ◽

JUN WU ◽

GELEI XIAO ◽

LEI HUO

Keyword(s):

Cell Invasion ◽

Glioma Cell ◽

Glioma Cells

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Effect of Geldanamycin on the Glioma Cell Invasion Induced by Hepatocyte Growth Factor

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.1033-1034.224 ◽

2014 ◽

Vol 1033-1034 ◽

pp. 224-228

Author(s):

Yan Xia Sun ◽

Guang Yu Zhou ◽

Li Li ◽

Xue Mei Han

Keyword(s):

Growth Factor ◽

Hepatocyte Growth Factor ◽

Cell Invasion ◽

Culture System ◽

Glioma Cell ◽

Glioma Cells ◽

Glioma Cell Line ◽

Invasive Growth ◽

Transwell Culture ◽

Hepatocyte Growth

To investigate the effect of geldanamycin (GDM) on the invasion ability of glioma cell induced by hepatocyte growth factor (HGF). Malignant glioma cell line U251-MG and U87-MG were cultured’and the capability of cell invasion was detected using a Transwell culture system. HGF significantly promoted the invasion ability of both U251-MG and U87-MG cells as compared with the normal control (NC) (P < 0.05). Forty eight hours after GDM treatment, the invasive growth of glioma cells was significantly decreased as compared with either NC or HGF group (P < 0.05). When cells were exposed to GDM plus HGF for 48 h, the cell invasion capability was greatly reduced as compared with either NC or HGF group (P < 0.05). The number of invaded cells in GDM plus HGF group was similar to that of GDM group.GDM can inhibit the invasion ability of glioma cells induced by HGF.

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MTSS1 is epigenetically regulated in glioma cells and inhibits glioma cell motility

Translational Oncology ◽

10.1016/j.tranon.2016.11.006 ◽

2017 ◽

Vol 10 (1) ◽

pp. 70-79 ◽

Cited By ~ 3

Author(s):

Daniel Luxen ◽

Gerrit H. Gielen ◽

Anke Waha ◽

Lukas Isselstein ◽

Tim Müller ◽

...

Keyword(s):

Cell Motility ◽

Glioma Cell ◽

Glioma Cells

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TAMI-04. TUMOR TREATING FIELDS (TTFIELDS) HINDER GLIOMA CELL MOTILITY THROUGH REGULATION OF MICROTUBULE AND ACTIN DYNAMICS

Neuro-Oncology ◽

10.1093/neuonc/noaa215.893 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii213-ii213

Author(s):

Tali Voloshin ◽

Rosa Schneiderman ◽

Alexandra Volodin ◽

Reuben Shamir ◽

Noa Kaynan ◽

...

Keyword(s):

Cell Motility ◽

Glioma Cell ◽

Therapeutic Potential ◽

Adhesion Formation ◽

Glioma Cells ◽

Microtubule Assembly ◽

Actin Dynamics ◽

Treatment Modalities ◽

Cellular Polarity ◽

Tumor Treating Fields

Abstract The ability of glioma cells to invade adjacent brain tissue remains a major obstacle to therapeutic disease management. Therefore, the development of novel treatment modalities that disrupt glioma cell motility could facilitate greater disease control. Tumor Treating Fields (TTFields), encompassing alternating electric fields within the intermediate frequency range, is an anticancer treatment delivered to the tumor region through transducer arrays placed non-invasively on the skin. This novel loco-regional treatment has demonstrated efficacy and safety and is FDA-approved in patients with glioblastoma and malignant pleural mesothelioma. TTFields are currently being investigated in other solid tumors in ongoing trials, including the phase 3 METIS trial (brain metastases from NSCLC; NCT02831959). Although established as an anti-mitotic treatment, the anti-metastatic potential of TTFields and its effects on cytoskeleton rapid dynamics during cellular motility warrant further investigation. Previous studies have demonstrated that TTFields inhibits metastatic properties of cancer cells. Identification of a unifying mechanism connecting the versatile TTFields-induced molecular responses is required to optimize the therapeutic potential of TTFields. In this study, confocal microscopy, computational tools, and biochemical analyses were utilized to show that TTFields disrupt glioma cellular polarity by interfering with microtubule assembly and directionality. Under TTFields application, changes in microtubule organization resulted in activation of GEF-H1, which led to an increase in active RhoA levels and consequent focal adhesion formation with actin cytoskeleton architectural changes. Furthermore, the optimal TTFields frequency for inhibition of invasion in glioma cells was 300 kHz, which differed from the optimal anti-mitotic frequency leading to glioma cell death of 200 kHz. The inhibitory effect of TTFields on migration was observed at fields intensities of 0.6 V/cm RMS (below the threshold of 1 V/cm RMS previously reported for cytotoxic effects). Together, these data identify discrete TTFields effects that disrupt processes crucial for glioma cell motility.

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