Cell‐Derived Vesicles as TRPC1 Channel Delivery Systems for the Recovery of Cellular Respiratory and Proliferative Capacities

Abstract

Pulsed electromagnetic fields (PEMFs) are capable of specifically activating a TRPC1‐mitochondrial axis underlying cell expansion and mitohormetic survival adaptations. This study characterizes cell‐derived vesicles (CDVs) generated from C2C12 murine myoblasts and shows that they are equipped with the sufficient molecular machinery to confer mitochondrial respiratory capacity and associated proliferative responses upon their fusion with recipient cells. CDVs derived from wild type C2C12 myoblasts include the cation‐permeable transient receptor potential (TRP) channels, TRPC1 and TRPA1, and directly respond to PEMF exposure with TRPC1‐mediated calcium entry. By contrast, CDVs derived from C2C12 muscle cells in which TRPC1 has been genetically knocked‐down using CRISPR/Cas9 genome editing, do not. Wild type C2C12‐derived CDVs are also capable of restoring PEMF‐induced proliferative and mitochondrial activation in two C2C12‐derived TRPC1 knockdown clonal cell lines in accordance to their endogenous degree of TRPC1 suppression. C2C12 wild type CDVs respond to menthol with calcium entry and accumulation, likewise verifying TRPA1 functional gating and further corroborating compartmental integrity. Proteomic and lipidomic analyses confirm the surface membrane origin of the CDVs providing an initial indication of the minimal cellular machinery required to recover mitochondrial function. CDVs hence possess the potential of restoring respiratory and proliferative capacities to senescent cells and tissues.

Pulsed electromagnetic fields synergize with graphene to enhance dental pulp stem cell-derived neurogenesis by selectively targeting TRPC1 channels

“…The sum of the presented results corroborated that TRPC1 is required for the developmental efficacy of this specific magnetic field paradigm applied to hDPSCs -as previously reported in skeletal myoblasts (Yap et al, 2019), MSCs (Parate et al, 2017;2020), and reconstituted cell systems (Kurth et al, 2020) -which, moreover, could be enhanced with the implementation of graphene substrates.…”

“…Despite TRPC1 and TRPM7 being the most ubiquitously expressed of all TRP channels (Jang et al, 2012), genetic silencing of TRPC1, but not TRPM7, precluded response to PEMF exposure (Yap et al, 2019). Moreover, vesicular delivery of TRPC1 is necessary and sufficient to restore responsiveness to magnetic field exposure in TRPC1 knock-down muscle cells (Kurth et al, 2020). Accordingly, adjusting the timing of PEMF exposure to coincide with the developmental window when TRPC1 was most prominent gave the greatest enhancement in hDPSC-derived neurogenesis and was preferentially conferred by 2DG substrates.…”

“…TRPC1 is necessary and sufficient to confer developmental sensitivity to brief exposure PEMFs (Kurth et al, 2020;Yap et al, 2019). Therefore, whether PEMF exposure could enhance neurogenesis was tested by targeting the developmental expression of TRPC1 conferred by 2DG.…”

“…1d), reinforcing that neurogenic induction was stronger on 2DG. PEMFs activate a TRPC1-mitochondrial axis promoting in vitro proliferation and subsequent differentiation (Kurth et al, 2020;Yap et al, 2019). Thus, the ability of 2DG in combination with PEMF exposure to augment the production of ROS and superoxide, products of cellular respiration, was investigated.…”

“…Specifically, PEMF-mediated activation of TRPC1 stimulates mitochondrial respiration and consequent ROS production upstream of eliciting mitochondrial survival adaptations following a process of magnetic mitohormesis (Yap et al, 2019). Indeed, vesicular delivery of TRPC1 alone to TRPC1 knockdown cells is sufficient to restore lost magnetosensitivity by reinstating mitochondrial responses to magnetic stimulation (Kurth et al, 2020). Similarly, PEMFs promote neurite outgrowth and differentiation in embryonic neural stem cells by recruiting the activity of TRPC1 channels (Ma et al, 2016).…”