TRPV4 activates the Cdc42/N-wasp pathway to promote glioblastoma invasion by altering cellular protrusions

Abstract

The invasion ability of glioblastoma (GBM) causes tumor cells to infiltrate the surrounding brain parenchyma and leads to poor outcomes. Transient receptor potential vanilloid 4 (TRPV4) exhibits a remarkable role in cancer cell motility, but the contribution of TRPV4 to glioblastoma metastasis is not fully understood. Here, we reported that TRPV4 expression was significantly elevated in malignant glioma compared to normal brain and low-grade glioma, and TRPV4 expression was negatively correlated with the prognosis of glioma patients. Functionally, stimulation of TRPV4 promoted glioblastoma cell migration and invasion, and repression of TRPV4 hindered the migration and invasion of glioblastoma cells in vitro. Molecularly, TRPV4 strongly colocalized and interacted with skeletal protein-F-actin at cellular protrusions, and TRPV4 regulated the formation of invadopodia and filopodia in glioblastoma cells. Furthermore, the Cdc42/N-wasp axis mediated the effect of TRPV4regulated cellular protrusions and invasion. Foremost, TRPV4 inhibitor treatment or downregulation of TRPV4 significantly reduced the invasion-growth of subcutaneously and intracranially transplanted glioblastoma in mice. In conclusion, the TRPV4/Cdc42/wasp signaling axis regulates cellular protrusion formation in glioblastoma cells and influences the invasion-growth phenotype of glioblastoma in vivo. TRPV4 may serve as a prognostic factor and specific therapeutic target for GBM patients. Abbreviations Cdc42 Cell division cycle 42 N-wasp Neural Wiskott-Aldrich syndrome proteins mDIA Mammalian homolog of diaphanous WAVE Wiskott-Aldrich syndrome protein family verprolin homologous protein CHO cell Chinese hamster ovary cell DRG cell Dorsal root ganglia cell RTVP-1 Related to testis-specific, vespid and pathogenesis protein 1 Glioblastoma (GBM) is the most common primary malignant tumor in the central nervous system (CNS). The current treatment protocol for glioblastoma patients is surgery and radiation therapy followed by treatment with temozolomide. Despite the development of multimodal treatment, the median life expectancy for patients with GBM is still approximately 14 months, and fewer than 5% of patients survive beyond 5 years of diagnosis 1. Unlike other malignant solid cancers, glioblastoma cells prefer to invade diffusely rather than create systemic metastases. The aggressive feature of glioblastoma cells is their invasion of surrounding normal brain tissues, which is the primary reason for incomplete surgical resection, chemo-radiation resistance and inevitable recurrence. There are very limited possibilities to target these processes therapeutically 2. In light of the poor outcome from current therapies in glioblastoma patients, a better understanding of GBM invasion may provide more effective interventions to manage this devastating disease.

Neuropathy-causing TRPV4 mutations disrupt TRPV4-RhoA interactions and impair neurite extension

“…5a, b ), but no activation of Rac1 and a small but significant activation of Cdc42 (Supplementary Fig. 5c, e ), consistent with a recent report 53 . To compare the spatial and temporal dynamics of RhoA activation in response to WT or mutant TRPV4 ion channel stimulation, we transfected T-Rex-TRPV4 cells with a FRET-based RhoA biosensor that displays increased FRET levels when RhoA is in the active, GTP-bound state 54 .…”

A mechanosensing mechanism mediated by IRSp53 controls plasma membrane shape homeostasis at the nanoscale

“…To discriminate between these mechanisms, we treated IRSp53 -/-R cells with different inhibitors and examined evagination resorption after compression. First, cell treatment with 10 μM of the N-WASP inhibitor Wiskostatin (73) reduced filopodia number as expected (74) (Supp. Figs.…”

Mechanisms of Invasion in Glioblastoma: Extracellular Matrix, Ca2+ Signaling, and Glutamate

“…TRP-vanilloid 4 (TRPV4), another TRP channel subfamily, is abnormally up-regulated in glioma, promoting migration and invasion through AKT/Rac1 signaling ( Ou-Yang et al, 2018 ). Stimulation of TRPV4 increases migration and invasion through the Cdc42/N-wasp axis by regulating cellular protrusions ( Yang et al, 2020 ). Study of various proteins interacting with TRPV2 in GBM may provide possible biomarkers for GBM diagnosis and lead to novel therapeutics ( Donate-Macian et al, 2018 ).…”