TRPM8-Rap1A Interaction Sites as Critical Determinants for Adhesion and Migration of Prostate and Other Epithelial Cancer Cells.

Emerging evidence indicates that the TRPM8 channel plays an important role in prostate cancer (PCa) progression, by impairing the motility of these cancer cells. Here, we reveal a novel facet of PCa motility control via direct protein-protein interaction (PPI) of the channel with the small GTPase Rap1A. The functional interaction of the two proteins was assessed by active Rap1 pull-down assays and live-cell imaging experiments. Molecular modeling analysis allowed the identification of four putative residues involved in TRPM8-Rap1A interaction. Point mutations of these sites impaired PPI as shown by GST-pull-down, co-immunoprecipitation, and PLA experiments and revealed their key functional role in the adhesion and migration of PC3 prostate cancer cells. More precisely, TRPM8 inhibits cell migration and adhesion by trapping Rap1A in its GDP-bound inactive form, thus preventing its activation at the plasma membrane. In particular, residues E207 and Y240 in the sequence of TRPM8 and Y32 in that of Rap1A are critical for the interaction between the two proteins not only in PC3 cells but also in cervical (HeLa) and breast (MCF-7) cancer cells. This study deepens our knowledge of the mechanism through which TRPM8 would exert a protective role in cancer progression and provides new insights into the possible use of TRPM8 as a new therapeutic target in cancer treatment.

H3.3K27M Mutation Controls Cell Growth and Resistance to Therapies in Pediatric Glioma Cell Lines.

High-grade gliomas represent the most lethal class of pediatric tumors, and their resistance to both radio- and chemotherapy is associated with a poor prognosis. Recurrent mutations affecting histone genes drive the tumorigenesis of some pediatric high-grade gliomas, and H3K27M mutations are notably characteristic of a subtype of gliomas called DMG (Diffuse Midline Gliomas). This dominant negative mutation impairs H3K27 trimethylation, leading to profound epigenetic modifications of genes expression. Even though this mutation was described as a driver event in tumorigenesis, its role in tumor cell resistance to treatments has not been deciphered so far. To tackle this issue, we expressed the H3.3K27M mutated histone in three initially H3K27-unmutated pediatric glioma cell lines, Res259, SF188, and KNS42. First, we validated these new H3.3K27M-expressing models at the molecular level and showed that K27M expression is associated with pleiotropic effects on the transcriptomic signature, largely dependent on cell context. We observed that the mutation triggered an increase in cell growth in Res259 and SF188 cells, associated with higher clonogenic capacities. Interestingly, we evidenced that the mutation confers an increased resistance to ionizing radiations in Res259 and KNS42 cells. Moreover, we showed that H3.3K27M mutation impacts the sensitivity of Res259 cells to specific drugs among a library of 80 anticancerous compounds. Altogether, these data highlight that, beyond its tumorigenic role, H3.3K27M mutation is strongly involved in pediatric glioma cells' resistance to therapies, likely through transcriptomic reprogramming.

Evofosfamide Is Effective against Pediatric Aggressive Glioma Cell Lines in Hypoxic Conditions and Potentiates the Effect of Cytotoxic Chemotherapy and Ionizing Radiations.

Hypoxia is a hallmark of many solid tumors and is associated with resistance to anticancer treatments. Hypoxia-activated prodrugs (HAPs) were developed to target the hypoxic regions of these tumors. Among 2nd generation HAPs, Evofosfamide (Evo, also known as TH-302) exhibits preclinical and clinical activities against adult glioblastoma. In this study, we evaluated its potential in the field of pediatric neuro-oncology. We assessed the efficacy of Evo in vitro as a single drug, or in combination with SN38, doxorubicin, and etoposide, against three pediatric high-grade glioma (pHGG) and three diffuse intrinsic pontine glioma (DIPG) cell lines under hypoxic conditions. We also investigated radio-sensitizing effects using clonogenic assays. Evo inhibited the growth of all cell lines, mainly under hypoxia. We also highlighted a significant synergism between Evo and doxorubicin, SN38, or etoposide. Finally, Evo radio-sensitized the pHGG cell line tested, both with fractionated and single-dose irradiation schedules. Altogether, we report here the first preclinical proof of evidence about Evofosfamide efficiency against hypoxic pHGG and DIPG cells. Since such tumors are highly hypoxic, and Evo potentiates the effect of ionizing radiation and chemotherapy, it appears as a promising therapeutic strategy for children with brain tumors.

[The art of war as applied to pediatric gliomas: Know your enemy]. / L'art de la guerre appliqué aux DIPG - Connais ton ennemi.

Pediatric brain cancers represent the most frequent solid tumors and the leading cause of cancer-driven mortality in children. Pediatric High Grade Gliomas display a very poor prognosis. Among these, DIPG (Diffuse Intrinsic Pontine Gliomas), localized to the brain stem, cannot benefit from a total exeresis due to this critical location and to their highly infiltrating nature. Radiotherapy remains the standard treatment against these tumors for almost five decades, and attempts to improve the prognosis of patients with chemotherapy or targeted therapies have failed. Thanks to the rise of high throughput sequencing, the knowledge of molecular alterations in pediatric gliomas strongly progressed and allowed to highlight distinct biomolecular entities and to establish more accurate diagnoses. In this review, we summarize this new information and the perspectives that it brings for clinical strategies.

TITLE: L'art de la guerre appliqué aux DIPG - Connais ton ennemi. ABSTRACT: Les tumeurs cérébrales pédiatriques représentent la principale cause de mortalité par cancer chez l'enfant. Alors que l'exérèse complète a une valeur pronostique dans certains gliomes de haut grade, les DIPG (diffuse intrinsic pontine gliomas) ne peuvent en bénéficier du fait d'une localisation critique au niveau du tronc cérébral et de leur caractère infiltrant. La radiothérapie demeure le traitement de référence contre ces tumeurs depuis bientôt cinquante ans, et les tentatives pour améliorer le pronostic vital des patients à l'aide de chimiothérapies ou de thérapies ciblées se sont révélées infructueuses. La connaissance des altérations moléculaires dans ces gliomes a fortement progressé cette dernière décennie, grâce aux progrès du séquençage à haut débit. Cela a permis de révéler des entités distinctes au niveau moléculaire et de préciser des diagnostics discriminants. Dans cette revue, nous faisons le point sur ces nouvelles connaissances et les perspectives qu'elles apportent en termes de stratégies cliniques.