Fusogenic Liposome-Mediated Extracellular Vesicles MicroRNA Profiling for Lung Cancer Diagnosis

Profiling circulating extracellular vesicle-associated microRNAs (EVs-miRNA) is essential for improving lung cancer (LC) diagnosis and prognosis in clinical tests. However, the complexity of RNA extraction procedures and the lack of an LC-specific EVs-miRNA signature limit the clinical applicability of the current liquid biopsy tools. Herein, we develop a fusogenic liposome nanoreactor for the direct profiling of plasma EVs-miRNAs. This platform integrates reagent-encapsulating liposomes, isothermal rolling circle amplification, and CRISPR-Cas12a endonuclease cleavage in a one-pot manner, allowing for ultrasensitive measurements down to fM levels. A LC-specific EVs-miRNA signature was identified through a designed four-phase screening procedure, which was validated by using both public databases and clinical samples. In a proof-of-concept cross-sectional and longitudinal study involving a clinical cohort ( n = 141), the profiled signature enabled early diagnosis, therapy monitoring, and prognosis evaluation with an accuracy of up to 93.1%. The platform streamlines the diagnostic workflow into a single fusogenic step, providing an RNA extraction-free and proof-of-concept liquid biopsy framework for potential LC management.