Separation-Free Extracellular Vesicle Microribonucleic Acid Profiling Using Structurally Oriented Membrane Fusion and...

Area of Science:

• Biochemistry
• Molecular Biology
• Nanotechnology

Background:

• Current methods for extracellular vesicle (EV) microRNA (miRNA) profiling are complex and have limitations in sensitivity or efficiency.
• Accurate and rapid detection of EV-derived miRNAs is crucial for early cancer diagnosis.

Purpose of the Study:

• To develop a novel, rapid, and sensitive platform for *in situ* detection of multiple tumor-specific EV-derived miRNAs.
• To overcome the limitations of existing EV miRNA profiling techniques.

Main Methods:

• A programmable, aptamer-mediated membrane fusion strategy was developed using Dual-color, Bone-shaped Biosensor-loaded Aptamer-Liposomes (DBS-Alip).
• DBS-Alip selectively fuses with intact EVs, delivering biosensors for nonenzymatic signal amplification within 30 minutes.
• The platform enables isolation-free, multiplexed detection of miRNAs directly from plasma.

Main Results:

• The DBS-Alip platform achieved ultralow detection limits (0.5 fM) with high sensitivity and specificity, requiring only 5 μL of plasma.
• Clinical validation demonstrated 100% accuracy in distinguishing early-stage nonsmall cell lung cancer patients from healthy donors and 90% accuracy from pneumonia cases.
• The method eliminates the need for RNA extraction or amplification.

Conclusions:

• The DBS-Alip platform provides a robust, programmable, and efficient EV-based liquid biopsy strategy.
• This approach holds significant potential for early cancer diagnosis through sensitive and specific miRNA detection.