Mechanoactivation of NOX2-generated ROS elicits persistent TRPM8 Ca ²⁺ signals that are inhibited by oncogenic KRas

Abstract

Changes in the mechanical microenvironment and mechanical signals are observed during tumor progression, malignant transformation, and metastasis. In this context, understanding the molecular details of mechanotransduction signaling may provide unique therapeutic targets. Here, we report that normal breast epithelial cells are mechanically sensitive, responding to transient mechanical stimuli through a two-part calcium signaling mechanism. We observed an immediate, robust rise in intracellular calcium (within seconds) followed by a persistent extracellular calcium influx (up to 30 min). This persistent calcium was sustained via microtubule-dependent mechanoactivation of NADPH oxidase 2 (NOX2)-generated reactive oxygen species (ROS), which acted on transient receptor potential cation channel subfamily M member 8 (TRPM8) channels to prolong calcium signaling. In contrast, the introduction of a constitutively active oncogenic KRas mutation inhibited the magnitude of initial calcium signaling and severely blunted persistent calcium influx. The identification that oncogenic KRas suppresses mechanically-induced calcium at the level of ROS provides a mechanism for how KRas could alter cell responses to tumor microenvironment mechanics and may reveal chemotherapeutic targets for cancer. Moreover, we find that expression changes in both NOX2 and TRPM8 mRNA predict poor clinical outcome in estrogen receptor (ER)-negative breast cancer patients, a population with limited available treatment options. The clinical and mechanistic data demonstrating disruption of this mechanically-activated calcium pathway in breast cancer patients and by KRas activation reveal signaling alterations that could influence cancer cell responses to the tumor mechanical microenvironment and impact patient survival.

Prognostic Value and Immune Infiltration of ARMC10 in Pancreatic Adenocarcinoma Via Integrated Bioinformatics Analyses

“…Several studies have illustrated the role of glutamate receptor in the pathophysiology mechanisms underlying multiple cancers, including breast cancer, glioblastoma, squamous cell lung cancer, prostate cancer, bladder cancer and melanoma [30][31][32][33][34][35]. Besides, researches have demonstrated the critical role of cation channels in the progression of various type of cancer, especially transient receptor potential cation channel subfamily V member 4 (TRPV4) [36-38], transient receptor potential cation channel subfamily M member 7/8 (TRPM7/8) [39][40][41][42] and two-pore channel 2 (TPC2) [43,44]. TPC2 is one of the cation channels located on endolysosomal membranes that could speed up movement between organelles and regulate phagocytosis, autophagy and lysosomal exocytosis [43].…”

Molecular Characterization of Membrane Steroid Receptors in Hormone-Sensitive Cancers

“…In this regard, TRPM8 has been reported to activate Akt and Glycogen Synthase Kinase 3 beta (GSK-3β) pathway to promote EMT and breast cancer aggressiveness [ 61 ] and to regulate proliferation, migration, and autophagy by activating AMP-Activated Protein Kinase (AMPK) and Unc-51 Like Autophagy Activating Kinase (ULK1) pathway [ 62 ] ( Figure 1 ). Furthermore, in vitro and clinical data support the role of TRPM8 as a biomarker for poor clinical outcome prediction in estrogen receptor (ER)-negative breast cancer patients [ 63 , 64 ].…”

Biophysics and Modeling of Mechanotransduction in Neurons: A Review

“…Based on how they react to mechanical stimuli, mechanosensitive TRP channels can be divided into two main categories: the mechanically-gated (MG) channels and the mechanically sensitive channels [129]. The first class includes the force sensors themselves that are directly activated by the stimulus [130,131], while the others are activated by second messengers whose release is triggered by the activation of the primary force sensor [128][129][130][131][132][133][134].…”

“…DAG occupies significantly lower space in the membrane than PIP 2 ; thus, PIP 2 hydrolysis might induce modifications of the membrane curvature in the proximity of the TRP channel leading to its mechanical activation [128]. In other cases, the indirect activation of TRP channels might be realized through the binding of specific ligands, such as DAG, PIP 2 [131], reactive oxygen species (ROS) [134], or serotonin [155].…”