mTORC1 induces plasma membrane depolarization and promotes preosteoblast senescence by regulating the sodium channel Scn1a

Senescence impairs preosteoblast expansion and differentiation into functional osteoblasts,blunts their responses to bone formation-stimulating factors and stimulates their secretion of osteoclast-activating factors.Due to these adverse effects,preosteoblast senescence is a crucial target for the treatment of age-related bone loss;however,the underlying mechanism remains unclear.We found that mTORC1 accelerated preosteoblast senescence in vitro and in a mouse model.Mechanistically,mTORC1 induced a change in the membrane potential from polarization to depolarization,thus promoting cell senescence by increasing Ca^(2+)influx and activating downstream NFAT/ATF3/p53 signaling.We further identified the sodium channel Scn1a as a mediator of membrane depolarization in senescent preosteoblasts.Scn1a expression was found to be positively regulated by mTORC1 upstream of C/EBPα,whereas its permeability to Na^(+)was found to be gated by protein kinase A(PKA)-induced phosphorylation.Prosenescent stresses increased the permeability of Scn1a to Na^(+)by suppressing PKA activity and induced depolarization in preosteoblasts.Together,our findings identify a novel pathway involving mTORC1,Scn1a expression and gating,plasma membrane depolarization,increased Ca^(2+)influx and NFAT/ATF3/p53 signaling in the regulation of preosteoblast senescence.Pharmaceutical studies of the related pathways and agents might lead to novel potential treatments for agerelated bone loss.