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 TRPC1mitochondrial 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 had 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.

Modulated TRPC1 expression predicts sensitivity of breast cancer to doxorubicin and magnetic field therapy: segue towards a precision medicine approach

“…Our results are in general agreement with previous studies drawing a correlation between TRPC1 expression and the malignancy status of several forms of cancer (17,19,34). Magnetic-sensitivity and downstream mitochondrial activation was previously shown to be specifically correlated with TRPC1 developmental and genetic expression and function (5,9,27,41), whereas the expression of other TRP channels did not show such a strong correlation (5,27,41) and genetic silencing of TRPM7 was unable to preclude magnetic sensitivity (5).…”

“…Mitohormesis describes a developmental process whereby low levels of oxidative stress instill mitochondrial survival adaptations by augmenting a cell's antioxidant defenses, whereas exaggerated elevations in ROS overwhelm a cell's existing antioxidant defenses to instead stymie cell survival (7). TRPC1 function was shown to be necessary and sufficient to confer mitochondrial responses to magnetic fields, ultimately invoking a novel process of magnetic mitohormesis (9). PEMF exposure undermines MCF-7 cell growth (Fig.…”

“…The magnetic sensitivity conferred by TRPC1 and its relevance to mitohormetic survival mechanisms make it a valuable target for clinical exploitation in cancer treatment (5,9,27).…”

“…CC-BY-NC-ND 4.0 International license made available under a (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is The copyright holder for this preprint this version posted April 30, 2021. ; https://doi.org/10.1101/2021.04.30.442085 doi: bioRxiv preprint Transient Receptor Potential Channel 1 (TRPC1) expression is necessary and sufficient to bestow PEMF-stimulated mitochondrial respiration and proliferation (9). Evidence of a TRPC1-mitochondrial axis exists with the findings that calcium entry modulates mitochondrial respiration (10), whereas mitochondrial ROS reciprocally modulates TRPC1 function (11).…”

Calcium‐Mediated Liposome Fusion to Engineer Giant Lipid Vesicles with Cytosolic Proteins and Reconstituted Mammalian Proteins

“…We believe that this system can be used to engineer fully functional hybrid liposomes, as we know from previous studies with CDVs that the functionality of the membrane proteins is preserved in the CDVs. [ 31,40 ] While we here showed the functionality of the cytosolic protein only, future studies will focus on functionality tests of the reconstituted membrane proteins. In case of the modified rtk, the functionality could be tested by exploiting the receptor kinase activity.…”