Multimodal Single Extracellular Vesicle Profiling Technologies to Unveil Clinical Heterogeneity

Extracellular vesicles (EVs) play significant roles in disease progression through their molecular cargos, including proteins, transcriptomes, and metabolites. However, the inherent heterogeneity of EVs in body fluids, originating from various cells with different functions, has impeded the translation of their biological significance to clinical applications, such as disease diagnosis, prognosis, and therapeutics. To address this heterogeneity issue, single EV profiling technologies have emerged in recent years. Despite increasing interest in multiomics profiling of EVs, single EV profiling has primarily been limited to qualitative or semiquantitative surface protein analysis due to technological constraints. In this review, we summarize qualitative and semiquantitative surface protein profiling methods and then highlight recent advances in multimodal single EV profiling platforms, capable of digitally quantifying surface proteins, detecting internal transcriptome, and simultaneously profiling surface protein and internal transcriptome. Furthermore, we discuss the outlook and challenges for multiomics single EV profiling including the technological hurdles to detect small molecules.