Detection of mutant cancer-derived circRNAs in extracellular vesicles by rolling circle amplification

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

doi:10.1016/j.omtn.2026.102970

Detection of mutant cancer-derived circRNAs in extracellular vesicles by rolling circle amplification

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

Massachusetts General Hospital
Harvard University
University of Medical Sciences Poznan
University of Padua
University of Basel
University of Milan - Bicocca
Partners HealthCare
Brigham and Women’s Hospital

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Article number	102970
Journal	Molecular Therapy Nucleic Acids
Volume	37
Issue number	3
DOIs	https://doi.org/10.1016/j.omtn.2026.102970
State	Published - 8 Sep 2026

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

cancer biomarkers
cancer mutations
circRNAs
extracellular vesicles
MT: non-coding RNAs
rolling circle amplification

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10.1016/j.omtn.2026.102970

Cite this

Zaborowski, M. P., Lai, C. P. K., Sammarco, A., György, B., Zhang, X., Lee, K., Na, Y. J., Ceppi, L., Breyne, K., Champagne, E. M., Cardini, J., Wesselhoeft, R. A., Iwanicki, M., Weissleder, R., Birrer, M. J., Lee, H., Tannous, B. A., & Breakefield, X. O. (2026). Detection of mutant cancer-derived circRNAs in extracellular vesicles by rolling circle amplification. Molecular Therapy Nucleic Acids, 37(3), Article 102970. https://doi.org/10.1016/j.omtn.2026.102970

@article{c7264c3679d444438a94fa7554f320d6,

title = "Detection of mutant cancer-derived circRNAs in extracellular vesicles by rolling circle amplification",

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.",

keywords = "cancer biomarkers, cancer mutations, circRNAs, extracellular vesicles, MT: non-coding RNAs, rolling circle amplification",

author = "Zaborowski, \{Miko{\l}aj Piotr\} and Lai, \{Charles Pin Kuang\} and Alessandro Sammarco and Bence Gy{\"o}rgy and Xuan Zhang and Kyungheon Lee and Na, \{Young Jeong\} and Lorenzo Ceppi and Koen Breyne and Champagne, \{Elizabeth Marie\} and Jack Cardini and Wesselhoeft, \{Robert Alexander\} and Marcin Iwanicki and Ralph Weissleder and Birrer, \{Michael J.\} and Hakho Lee and Tannous, \{Bakhos A.\} and Breakefield, \{Xandra O.\}",

note = "Publisher Copyright: {\textcopyright} 2026 The Authors.",

year = "2026",

month = sep,

day = "8",

doi = "10.1016/j.omtn.2026.102970",

language = "English",

volume = "37",

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Zaborowski, MP, Lai, CPK, Sammarco, A, György, B, Zhang, X, Lee, K, Na, YJ, Ceppi, L, Breyne, K, Champagne, EM, Cardini, J, Wesselhoeft, RA, Iwanicki, M, Weissleder, R, Birrer, MJ, Lee, H, Tannous, BA & Breakefield, XO 2026, 'Detection of mutant cancer-derived circRNAs in extracellular vesicles by rolling circle amplification', Molecular Therapy Nucleic Acids, vol. 37, no. 3, 102970. https://doi.org/10.1016/j.omtn.2026.102970

TY - JOUR

T1 - Detection of mutant cancer-derived circRNAs in extracellular vesicles by rolling circle amplification

AU - Zaborowski, Mikołaj Piotr

AU - Lai, Charles Pin Kuang

AU - Sammarco, Alessandro

AU - György, Bence

AU - Zhang, Xuan

AU - Lee, Kyungheon

AU - Na, Young Jeong

AU - Ceppi, Lorenzo

AU - Breyne, Koen

AU - Champagne, Elizabeth Marie

AU - Cardini, Jack

AU - Wesselhoeft, Robert Alexander

AU - Iwanicki, Marcin

AU - Weissleder, Ralph

AU - Birrer, Michael J.

AU - Lee, Hakho

AU - Tannous, Bakhos A.

AU - Breakefield, Xandra O.

PY - 2026/9/8

Y1 - 2026/9/8

N2 - 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.

AB - 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.

KW - cancer biomarkers

KW - cancer mutations

KW - circRNAs

KW - extracellular vesicles

KW - MT: non-coding RNAs

KW - rolling circle amplification

UR - https://www.scopus.com/pages/publications/105041912827

UR - https://www.scopus.com/pages/publications/105041912827#tab=citedBy

U2 - 10.1016/j.omtn.2026.102970

DO - 10.1016/j.omtn.2026.102970

M3 - Article

AN - SCOPUS:105041912827

SN - 2162-2531

VL - 37

JO - Molecular Therapy Nucleic Acids

JF - Molecular Therapy Nucleic Acids

IS - 3

M1 - 102970

ER -

Detection of mutant cancer-derived circRNAs in extracellular vesicles by rolling circle amplification

Abstract

UN SDGs

Keywords

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