Amplifying mutational profiling of extracellular vesicle mRNA with SCOPE

Area of Science:

• Biotechnology
• Molecular Biology
• Genomics

Background:

• Extracellular vesicles (EVs) in liquid biopsies contain messenger RNA (mRNA) valuable for clinical insights like somatic mutations and tumor recurrence.
• The low abundance of EV mRNA in liquid biopsies limits its clinical utility, necessitating large sample volumes or specialized techniques.
• Current methods struggle with sensitivity and resolution for comprehensive EV mRNA profiling.

Purpose of the Study:

• To introduce Self-amplified and CRISPR-aided Operation to Profile EVs (SCOPE), a novel platform for highly sensitive EV mRNA detection.
• To demonstrate SCOPE's capability for single-nucleotide resolution and sub-attomolar detection limits.
• To validate SCOPE's application in clinical oncology, including early cancer detection and patient stratification.

Main Methods:

• Leveraging CRISPR-Cas13 technology for target RNA recognition and signal amplification.
• Developing single-pot assays with optimized protocols for efficient EV mRNA analysis.
• Designing specific probes for key cancer-associated mutations (KRAS, BRAF, EGFR, IDH1) and implementing an automated detection device.

Main Results:

• Achieved sub-attomolar detection limits for EV mRNA with single-nucleotide resolution.
• Successfully detected early-stage lung cancer in animal models.
• Monitored tumor mutational burden in colorectal cancer patients and stratified glioblastoma patients.

Conclusions:

• SCOPE significantly enhances the sensitivity and efficiency of EV mRNA detection in liquid biopsies.
• The platform accelerates diagnostic readouts, paving the way for broader clinical adoption of EVs in precision oncology.
• SCOPE offers a powerful tool for non-invasive cancer diagnostics and personalized treatment strategies.