{"doi":"10.1002/jex2.108","title":"Integrating extracellular vesicle and circulating cell‐free DNA analysis using a single plasma aliquot improves the detection of HER2 positivity in breast cancer patients","abstract":"<jats:title>Abstract</jats:title>\n                  <jats:p>\n                    Multi‐analyte liquid biopsies represent an emerging opportunity for non‐invasive cancer assessment. We developed ONCE (One Aliquot for Circulating Elements), an approach for the isolation of extracellular vesicles (EV) and cell‐free DNA (cfDNA) from a single aliquot of blood. We assessed ONCE performance to classify HER2‐positive early‐stage breast cancer (BrCa) patients by combining EV‐associated RNA (EV‐RNA) and cfDNA signals on\n                    <jats:italic>n</jats:italic>\n                     = 64 healthy donors (HD) and non–metastatic BrCa patients. Specifically, we isolated EV‐enriched samples by a charge‐based (CB) method and investigated EV‐RNA and cfDNA by next‐generation sequencing (NGS) and by digital droplet PCR (ddPCR). Sequencing of cfDNA and EV‐RNA from HER2‐ and HER2+ patients demonstrated concordance with in situ molecular analyses of matched tissues. Combined analysis of the two circulating analytes by ddPCR showed increased sensitivity in ERBB2/HER2 detection compared to single nucleic acid components. Multi‐analyte liquid biopsy prediction performance was comparable to tissue‐based sequencing results from TCGA. Also, imaging flow cytometry analysis revealed HER2 protein on the surface of EV isolated from the HER2+ BrCa plasma, thus corroborating the potential relevance of studying EV as companion analyte to cfDNA. This data confirms the relevance of combining cfDNA and EV‐RNA for HER2 cancer assessment and supports ONCE as a valuable tool for multi‐analytes liquid biopsies’ clinical implementation.\n                  </jats:p>","journal":"Journal of Extracellular Biology","year":2023,"id":627871,"datarank":0.5814949248554534,"base_score":2.639057329615259,"endowment":2.639057329615259,"self_citation_contribution":0.3958585994422889,"citation_network_contribution":0.1856363254131645,"self_endowment_contribution":0.3958585994422889,"citer_contribution":0.1856363254131645,"corpus_percentile":null,"corpus_rank":null,"citation_count":13,"citer_count":12,"citers_with_citation_signal":11,"citers_with_endowment":11,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":77619,"name":"Yari Ciani","orcid":"0000-0002-9635-2830","position":1,"is_corresponding":false},{"id":1026676,"name":"Orsetta Quaini","orcid":"0009-0008-3466-8988","position":2,"is_corresponding":false},{"id":1625328,"name":"Simone Tomasini","orcid":null,"position":3,"is_corresponding":false},{"id":1625329,"name":"Michela Notarangelo","orcid":null,"position":4,"is_corresponding":false},{"id":1625330,"name":"Federico Vannuccini","orcid":null,"position":5,"is_corresponding":false},{"id":1625331,"name":"Alessia Marinelli","orcid":null,"position":6,"is_corresponding":false},{"id":1625332,"name":"Elena Leonardi","orcid":null,"position":7,"is_corresponding":false},{"id":1625333,"name":"Stefano Pontalti","orcid":null,"position":8,"is_corresponding":false},{"id":749268,"name":"Angela Martinelli","orcid":"0000-0001-6442-3190","position":9,"is_corresponding":false},{"id":849515,"name":"Daniele Rossetto","orcid":"0000-0002-9399-0888","position":10,"is_corresponding":false},{"id":1125646,"name":"Isabella Pesce","orcid":null,"position":11,"is_corresponding":false},{"id":849516,"name":"Sheref S. Mansy","orcid":"0000-0003-2382-198X","position":12,"is_corresponding":false},{"id":414126,"name":"Mattia Barbareschi","orcid":"0000-0003-0217-3223","position":13,"is_corresponding":false},{"id":147337,"name":"Antonella Ferro","orcid":null,"position":14,"is_corresponding":false},{"id":1027430,"name":"Orazio Caffo","orcid":null,"position":15,"is_corresponding":false},{"id":554640,"name":"Gerhardt Attard","orcid":"0000-0002-4811-7983","position":16,"is_corresponding":false},{"id":294390,"name":"Dolores Di Vizio","orcid":"0000-0002-9787-6556","position":17,"is_corresponding":false},{"id":215744,"name":"Vito G. D'Agostino","orcid":"0000-0003-3379-2254","position":18,"is_corresponding":false},{"id":215739,"name":"Caterina Nardella","orcid":null,"position":19,"is_corresponding":false},{"id":35285,"name":"Francesca Demichelis","orcid":"0000-0002-8266-8631","position":20,"is_corresponding":false},{"id":775438,"name":"Vera Mugoni","orcid":"0000-0001-6964-4179","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Integrating extracellular vesicle and circulating cell‐free DNA analysis using a single plasma aliquot improves the detection of HER2 positivity in breast cancer patients","abstract":"<jats:title>Abstract</jats:title>\n                  <jats:p>\n                    Multi‐analyte liquid biopsies represent an emerging opportunity for non‐invasive cancer assessment. We developed ONCE (One Aliquot for Circulating Elements), an approach for the isolation of extracellular vesicles (EV) and cell‐free DNA (cfDNA) from a single aliquot of blood. We assessed ONCE performance to classify HER2‐positive early‐stage breast cancer (BrCa) patients by combining EV‐associated RNA (EV‐RNA) and cfDNA signals on\n                    <jats:italic>n</jats:italic>\n                     = 64 healthy donors (HD) and non–metastatic BrCa patients. Specifically, we isolated EV‐enriched samples by a charge‐based (CB) method and investigated EV‐RNA and cfDNA by next‐generation sequencing (NGS) and by digital droplet PCR (ddPCR). Sequencing of cfDNA and EV‐RNA from HER2‐ and HER2+ patients demonstrated concordance with in situ molecular analyses of matched tissues. Combined analysis of the two circulating analytes by ddPCR showed increased sensitivity in ERBB2/HER2 detection compared to single nucleic acid components. Multi‐analyte liquid biopsy prediction performance was comparable to tissue‐based sequencing results from TCGA. Also, imaging flow cytometry analysis revealed HER2 protein on the surface of EV isolated from the HER2+ BrCa plasma, thus corroborating the potential relevance of studying EV as companion analyte to cfDNA. This data confirms the relevance of combining cfDNA and EV‐RNA for HER2 cancer assessment and supports ONCE as a valuable tool for multi‐analytes liquid biopsies’ clinical implementation.\n                  </jats:p>","is_dataset_classified":null,"base_score":2.639057329615259,"endowment":2.639057329615259,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"38046436","pmcid":"PMC10688391","openalex_id":"https://openalex.org/W4387077005","authors":[],"funders":[{"funder_name":"NCI NIH HHS","grant_id":"P50 CA211024","title":null},{"funder_name":"National Institutes of Health","grant_id":"1P50CA211024-01A1","title":"Weill Cornell Medicine (WCM) SPORE in Prostate Cancer"},{"funder_name":"Fondazione Cassa Di Risparmio Di Trento E Rovereto","grant_id":"","title":null}],"total_grants":3,"fwci":1.2056,"citation_percentile":0.78014344,"influential_citations":0,"citation_trend":[{"year":2024,"count":7},{"year":2025,"count":4},{"year":2026,"count":2}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/jex2.108","host_type":"journal"},{"url":"https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/jex2.108","host_type":"publisher"},{"url":"https://isevjournals.onlinelibrary.wiley.com/doi/pdf/10.1002/jex2.108","host_type":"publisher"},{"url":"https://isevjournals.onlinelibrary.wiley.com/doi/full-xml/10.1002/jex2.108","host_type":"publisher"},{"url":"https://doi.org/10.1002/jex2.108","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/38046436","host_type":"repository"},{"url":"https://isevjournals.onlinelibrary.wiley.com/doi/10.1002/jex2.108","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/10688391","host_type":"repository"},{"url":"https://doaj.org/article/4ec0396d0d6d4b53b34bf42dcddc3061","host_type":"repository"},{"url":"https://hdl.handle.net/11573/1763442","host_type":"repository"},{"url":"https://iris.unitn.it/bitstream/11572/427311/1/mugoni_barbareschi_2023_her2_breast_cancer.pdf","host_type":"repository"},{"url":"https://pmc.ncbi.nlm.nih.gov/articles/PMC10688391/pdf/JEX2-2-e108.pdf","host_type":"repository"},{"url":"http://dx.doi.org/10.1002/jex2.108","host_type":""},{"url":"https://www.scopus.com/pages/publications/85194002923","host_type":""},{"url":"https://vbn.aau.dk/da/publications/2373041c-29a6-4e7c-ad0e-987d637fe630","host_type":""},{"url":"https://hdl.handle.net/11572/427311","host_type":""}],"fields_of_study":["Extracellular vesicles in disease","Advanced biosensing and bioanalysis techniques","MicroRNA in disease regulation","0303 health sciences","03 medical and health sciences"],"mesh_terms":[],"keywords":["Liquid biopsy","Analyte","Cell-free fetal DNA","Breast cancer","RNA","Extracellular vesicle","Digital polymerase chain reaction","Nucleic acid","DNA","Concordance","Flow cytometry","Cancer","Cancer research","Oncology","Chemistry","Computational biology","Molecular biology","microRNA","Biology","Medicine","Internal medicine","Chromatography","Gene","Microvesicles","Polymerase chain reaction","Biochemistry","Genetics","Her2","Imagestream","Ddpcr","Extracellular Vesicles (Ev)","Once Approach","Ev‐rna","Cell‐free Dna (Cfdna)","QH573-671","breast cancer, cell-free DNA (cfDNA), ddPCR, EV-RNA, extracellular vesicles (EV), HER2, ImageStream, ONCE approach","breast cancer; cell-free DNA (cfDNA); ddPCR; EV-RNA; extracellular vesicles (EV); HER2; ImageStream; ONCE approach","EV-RNA","cell-free DNA (cfDNA)","Cytology","Research Articles"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Good health and well-being"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"gen"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-05T03:04:35.869777Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}