FRET imaging of glycoRNA on small extracellular vesicles enabling sensitive cancer diagnostics

Tingju Ren, Yingzhi Zhang, Yuxiao Tong, Qi Zhang, Tianhao Wang, Yue Wang, Chunguang Yang and Zhangrun Xu ()
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Tingju Ren: Northeastern University
Yingzhi Zhang: The First Affiliated Hospital of China Medical University
Yuxiao Tong: Northeastern University
Qi Zhang: Northeastern University
Tianhao Wang: Northeastern University
Yue Wang: Northeastern University
Chunguang Yang: Northeastern University
Zhangrun Xu: Northeastern University

Nature Communications, 2025, vol. 16, issue 1, 1-19

Abstract: Abstract Glycosylated RNAs (glycoRNAs), a recently discovered class of membrane-associated glyco-molecules, remain poorly understood in function and clinical value due to limited detection methods. Here, we show a dual recognition Förster resonance energy transfer (drFRET) strategy using nucleic acid probes to detect N-acetylneuraminic acid-modified RNAs, enabling sensitive, selective profiling of glycoRNAs on small extracellular vesicles (sEVs) from minimal biofluids (10 μl initial biofluid). Using drFRET, we identify 5 prevalent sEV glycoRNAs derived from 7 cancer cell lines. In a 100-patient cohort (6 cancer types and non-cancer controls), sEV glycoRNA profiles achieve 100% accuracy (95% confidence interval) in distinguishing cancers from non-cancer cases and 89% accuracy in classifying specific cancer types. Furthermore, drFRET reveal that sEV glycoRNAs specifically interact with Siglec proteins and P-selectin, which is critical for sEV cellular internalization. The drFRET strategy provides a versatile and sensitive platform for the imaging and functional analysis of sEV glycoRNAs, with promising implications for clinical applications.

Date: 2025
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DOI: 10.1038/s41467-025-58490-2

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