In Situ MicroRNA Profiling of Single Tumor Extracellular Vesicles for Precise Prostate Cancer Diagnosis

Area of Science:

• Biochemistry
• Molecular Biology
• Oncology

Background:

• Extracellular vesicles (EVs) contain stable microRNAs (miRNAs) reflecting cell characteristics, making them potential cancer biomarkers.
• Challenges in sensitive detection of tumor-EV miRNAs include EV heterogeneity and low abundance in early-stage cancer.

Purpose of the Study:

• To develop a sensitive and specific method for profiling miRNAs within tumor-derived extracellular vesicles (TEVs) for precise prostate cancer (PCa) diagnosis.
• To enable single-particle level analysis of TEVs for enhanced diagnostic accuracy.

Main Methods:

• A one-pot strategy, miR-nSTEV, utilizing dual-surface protein recognition (CD63, EpCAM) and DNA barcoding for TEV identification and fusion with Tag-Lipo probes.
• Nanoflow cytometry (nFCM) for precise miRNA analysis of individual TEVs.
• Profiling of six PCa-associated miRNAs in cell culture and clinical plasma samples.

Main Results:

• The miR-nSTEV assay achieved 100% accuracy in distinguishing PCa patients from healthy donors.
• Demonstrated superior performance compared to conventional RT-qPCR with a simplified workflow.
• Effectively eliminated interference from free nucleic acids and RNases, enhancing detection fidelity.

Conclusions:

• miR-nSTEV provides a robust, sensitive, and noninvasive tool for real-time TEV miRNA analysis.
• This method facilitates early detection and precision diagnostics for prostate cancer patients.
• Paves the way for advanced noninvasive cancer diagnostics using extracellular vesicle miRNAs.