Controlled release of exosomes by AMPK/SREBPs pathway

Exosome-like extracellular vesicles (EVs) are very important vesicles for intercellular signal transmission. Although host factors involving exosome-like EV production and release has been reported, the cellular signaling pathway regulating EV production is largely unknown. Here we performed a kinase inhibitor library screening to identify pathways controlling EV’s production. We found a kinase inhibitor, APY0201, which significantly increased EV secretion through upregulation of intracellular sterol synthesis and the volume of intracellular multivesicular bodies (MVBs). APY0201 specifically caused the decrease of AMP-activated protein kinase (AMPK) phosphorylation and activation of sterol regulatory element-binding protein (SREBP1), a pivotal transcription factor of sterol synthesis gene expression. In addition, both AMPK agonist and SREBP inhibitor deteriorate augmented EV production induced by APY0201. Ectopic expression of AMPK and SREBP1 corroborated that AMPK and SREBP1 are two major host factors governing the production of exosome-like EVs. Taken together, we identified that APY0201 is a novel kinase inhibitor targeting AMPK-SREBP1 pathway which is important for EV biogenesis and release. Our findings might pave the way to achievable finely upregulated EV production. RNA-seq of 8 cell samples, including 2 DMSO controls. Each sample had 2 biological replicates.