d689124d82fff9ddc140921ae01df499 pmc_10487825.pdf 2be535cdd9cb34488490b4e9c18bf94889bc37dd pmc_10487825.pdf a4b0d5527d01c36b2ca7bdca478ac6593444b6a3733fbc2cc6bc57ea30eeb8e7 pmc_10487825.pdf Title: Circulating miRNA Expression Is Inversely Correlated with Tumor Tissue or Sentinel Lymph Nodes in Estrogen Receptor-Positive Early Breast Cancer Patients Subject: The deregulation of microRNAs (miRNAs) is associated with the various steps of the metastatic process. In addition, circulating miRNAs are remarkably stable in peripheral blood, making them ideal noninvasive biomarkers for disease diagnosis. Here, we performed a proof-of-principle study to determine whether tumor-tissue-derived miRNAs are traceable to plasma in ER-positive early breast cancer patients. We performed RNA-sequencing on 30 patients for whom plasma, sentinel lymph nodes (SLNs) and tumor tissue were available. We carried out differential expression, gene ontology and enrichment analyses. Our results show that circulating miRNAs are inversely expressed compared with tumor tissue or SLNs obtained from the same patients. Our differential expression analysis shows the overall downregulation of circulating miRNAs. However, the expression of miR-643a-3p and miR-223 was up-regulated in patients with positive SLNs. Furthermore, gene ontology analysis showed the significant enrichment of biological processes associated with the regulation of epithelial cell proliferation and transcriptional regulation commonly involved in the promotion of metastases. Our results suggest the potential role of several circulating miRNAs as surrogate markers of lymph node metastases in early breast cancer patients. Further preclinical and clinical studies are required to understand the biological significance of the most significant miRNAs and to validate our results in a larger cohort of patients. Keywords: miRNAs; early breast cancer; sentinel lymph node; metastasis; biomarkers Author: Daniel Escuin, Laura López-Vilaró, Olga Bell, Josefina Mora, Bárbara García-Valdecasas, Antonio Moral, Montserrat Clos, Laia Boronat, Cristina Arqueros and Agustí Barnadas Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Sun Aug 27 10:26:40 2023 CEST ModDate: Sun Aug 27 10:30:50 2023 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 7082087 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- PRJKGQ+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 HCBGQO+VnURWPalladioL Type 1 Custom yes yes no 15 0 IODGLS+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 NHHNFB+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 TYPTQO+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 46 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 49 0 KXDJCR+CMSY10 Type 1 Builtin yes yes no 71 0 IFDCCP+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 93 0 IFDCEA+PalatinoLinotype CID TrueType Identity-H yes yes yes 96 0 IFDCEB+PalatinoLinotype TrueType WinAnsi yes yes no 102 0 IFDCED+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 105 0 IFDCAP+PalatinoLinotype,Bold CID TrueType Identity-H yes yes yes 148 0 IFDCCP+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 154 0 IFDCEA+PalatinoLinotype CID TrueType Identity-H yes yes yes 157 0 IFDCEB+PalatinoLinotype TrueType WinAnsi yes yes no 163 0 IFDCED+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 166 0 IFDEJG+PalatinoLinotype TrueType MacRoman yes yes no 169 0 WBVKDJ+URWPalladioL-BoldItal Type 1 Custom yes yes no 190 0 IFDCCP+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 204 0 IFDCEB+PalatinoLinotype TrueType WinAnsi yes yes no 207 0 IFDCED+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 210 0 IFDEFH+PalatinoLinotype,Bold TrueType MacRoman yes yes no 213 0 IFDEJG+PalatinoLinotype TrueType MacRoman yes yes no 216 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 230 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-05-07 04:23:11 CEST RepresentationInformation: pmc_10487825.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-04-24 02:39:36 CEST Size: 7082087 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 399 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results Destination: section.2 Children: Item: Title: Patients Destination: subsection.2.1 Item: Title: RNA-Sequencing Destination: subsection.2.2 Item: Title: Unsupervised Clustering Analysis Destination: subsection.2.3 Item: Title: Differentially Expressed Circulating miRNAs Destination: subsection.2.4 Item: Title: Biological Significance and Enriched Analysis of Circulating miRNAs Destination: subsection.2.5 Item: Title: Clinicopathological Correlation with Circulating miRNAs Destination: subsection.2.6 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Patients Destination: subsection.4.1 Item: Title: Sample Processing Destination: subsection.4.2 Item: Title: RNA Isolation Destination: subsection.4.3 Item: Title: RNA-Sequencing Destination: subsection.4.4 Item: Title: Differential Expression Analysis Destination: subsection.4.5 Item: Title: Biological Significance Analysis, Gene Ontology (GO) and Enrichment Analysis Destination: subsection.4.6 Item: Title: Statistics Destination: subsection.4.7 Item: Title: References Destination: section.A. Info: Title: Circulating miRNA Expression Is Inversely Correlated with Tumor Tissue or Sentinel Lymph Nodes in Estrogen Receptor-Positive Early Breast Cancer Patients Author: Daniel Escuin, Laura López-Vilaró, Olga Bell, Josefina Mora, Bárbara García-Valdecasas, Antonio Moral, Montserrat Clos, Laia Boronat, Cristina Arqueros and Agustí Barnadas Subject: The deregulation of microRNAs (miRNAs) is associated with the various steps of the metastatic process. In addition, circulating miRNAs are remarkably stable in peripheral blood, making them ideal noninvasive biomarkers for disease diagnosis. Here, we performed a proof-of-principle study to determine whether tumor-tissue-derived miRNAs are traceable to plasma in ER-positive early breast cancer patients. We performed RNA-sequencing on 30 patients for whom plasma, sentinel lymph nodes (SLNs) and tumor tissue were available. We carried out differential expression, gene ontology and enrichment analyses. Our results show that circulating miRNAs are inversely expressed compared with tumor tissue or SLNs obtained from the same patients. Our differential expression analysis shows the overall downregulation of circulating miRNAs. However, the expression of miR-643a-3p and miR-223 was up-regulated in patients with positive SLNs. Furthermore, gene ontology analysis showed the significant enrichment of biological processes associated with the regulation of epithelial cell proliferation and transcriptional regulation commonly involved in the promotion of metastases. Our results suggest the potential role of several circulating miRNAs as surrogate markers of lymph node metastases in early breast cancer patients. Further preclinical and clinical studies are required to understand the biological significance of the most significant miRNAs and to validate our results in a larger cohort of patients. 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