The identification of novel small extracellular vesicle (sEV) production modulators using luciferase?based sEV quantification method

Small extracellular vesicles (sEVs) are nano-sized secreted from various cells that contain bioactive metabolites and function as key regulators for intercellular communication. sEVs modulate diverse biological pathological processes in the body, amount of circulating has been reported to correlate with certain disease progression. Therefore, identification small molecular compounds can control sEV production may become a novel therapeutic strategy. In this study, rapid, highly sensitive quantification method utilizing fusion proteins consisting Gaussia luciferase (gLuc) reporter protein markers (CD63 CD82) was developed. A total 480 were screened identify potent inducers inhibitors gLuc activity. Two compounds, KPYC08425 KPYC12163, showed significant dose-dependent changes activity minimal cytotoxicity based on LDH assay. The efficacy these two further evaluated by isolated sEVs. Further evaluation KPYC12163 suggested autolysosomal pathway be involved its inhibitory effect production.