Detection of mutant cancer-derived circRNAs in extracellular vesicles by rolling circle amplification
Year: 2026
Type: article
Abstract: Circular RNAs (circRNAs) form during splicing as a closed ring and are more resistant to degradation than linear RNAs. Cells release circRNAs within membrane-bound extracellular vesicles (EVs), which transport DNA, RNA, and proteins. Whether circRNAs in EVs can be used as biomarkers remains uncertain. We aimed to identify mutant circRNAs in EVs to develop a new biomarker approach. We demonstrated that ovarian cancer cells release and enrich circRNAs in EVs. We detected mutant circular transcripts from TP53 , KRAS , and PIK3CA genes in EVs from cancer cells. We noticed that circRNAs in EVs undergo intense rolling circle amplification (RCA), producing multiple copies of the mutant sequence. We revealed that increased reverse transcription time and extended PCR elongation time intensified RCA. Using our RCA protocol, we detected mutant circRNAs derived from the TP53 gene in EVs from the plasma of ovarian cancer-bearing animals. In this study, we present a new model of mutant circRNA amplification from EVs, which may provide a basis for a diagnostic test for ovarian cancer patients.
Authors: Mikołaj Piotr Zaborowski, Charles Pin-Kuang Lai, Alessandro Sammarco, Bence György, Xuan Zhang, Kyungheon Lee, Young Jeong Na, Lorenzo Ceppi, Koen Breyne, Elizabeth Marie Champagne, Jack Cardini, Robert Alexander Wesselhoeft, Marcin Iwanicki, Ralph Weissleder, Michael J. Birrer, Hakho Lee, Bakhos A. Tannous, Xandra O. Breakefield
Institutions: Poznan University of Medical Sciences, Harvard University, Massachusetts General Hospital, Institute of Bioorganic Chemistry, Polish Academy of Sciences, University of Padua, University of Basel, Institute of Molecular and Clinical Ophthalmology Basel, Center for Systems Biology, University of Milano-Bicocca, Brigham and Women's Hospital, Mass General Brigham, Stevens Institute of Technology
Language: English
Topic: Circular RNAs in diseases
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,900
Funders: Changping Laboratory, American Heart Association, Kosciuszko Foundation, Massachusetts General Hospital, European Commission, Fundacja na rzecz Nauki Polskiej, Uniwersytet Medyczny im. Karola Marcinkowskiego w Poznaniu, National Institutes of Health, National Cancer Institute, Office of Strategic Coordination
Awards: Extracellular Vesicles (EVs) and Genes in the Biology and Therapy of Gliomas, G2302417481, The Role of VLDL-trafficked Small RNAs in Metabolic Disease, G3489812386, A Novel Paradigm for LDL-Associated Extracellular Small RNA in the Pathogenesis of Atherosclerosis, The power of extracellular vesicles in glioblastoma, G7370233133, G8088043049
OpenAlex ID: W7163080532