846e539b964944a409f104c9c29ed395 223895.pdf 0ff32ba0c544e1d323b4b22ea01a17da77fc3ebe 223895.pdf 02e0e3dfc662636804c796815f1d23c31216bef6767eea108c2c2221d0662be3 223895.pdf Title: Extracellular Vesicles-Based Biomarkers Represent a Promising Liquid Biopsy in Endometrial Cancer Subject: Tumor-derived extracellular vesicles (EVs) are secreted in large amounts into biological fluids of cancer patients. The analysis of EVs cargoes has been associated with patient´s outcome and response to therapy. However, current technologies for EVs isolation are tedious and low cost-efficient for routine clinical implementation. To explore the clinical value of circulating EVs analysis we attempted a proof-of-concept in endometrial cancer (EC) with ExoGAG, an easy to use and highly efficient new technology to enrich EVs. Technical performance was first evaluated using EVs secreted by Hec1A cells. Then, the clinical value of this strategy was questioned by analyzing the levels of two well-known tissue biomarkers in EC, L1 cell adhesion molecule (L1CAM) and Annexin A2 (ANXA2), in EVs purified from plasma in a cohort of 41 EC patients and 20 healthy controls. The results demonstrated the specific content of ANXA2 in the purified EVs fraction, with an accurate sensitivity and specificity for EC diagnosis. Importantly, high ANXA2 levels in circulating EVs were associated with high risk of recurrence and non-endometrioid histology suggesting a potential value as a prognostic biomarker in EC. These results also confirmed ExoGAG technology as a robust technique for the clinical implementation of circulating EVs analyses. Keywords: EVs; ExoGAG; endometrial cancer; ANXA2 Author: Carolina Herrero, Alexandre de la Fuente, Carlos Casas-Arozamena, Victor Sebastian, Martin Prieto, Manuel Arruebo, Alicia Abalo, Eva Colás, Gema Moreno-Bueno, Antonio Gil-Moreno, Ana Vilar, Juan Cueva, Miguel Abal and Laura Muinelo-Romay Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.18 CreationDate: Thu Dec 12 09:22:50 2019 CET ModDate: Thu Jul 29 14:13:10 2021 CEST 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: 3140479 bytes Optimized: yes PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- AIQCCL+VnURWPalladioL-Bold Type 1 Custom yes yes no 524 0 AZBFJT+VnURWPalladioL Type 1 Custom yes yes no 527 0 FDYFVT+URWPalladioL-Ital Type 1 Custom yes yes no 531 0 VCVIBQ+URWPalladioL-Bold Type 1 Custom yes yes no 534 0 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SignatureMatches: PDF-hul MIMEtype: application/pdf Profile: Linearized PDF PDFMetadata: Objects: 0 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Item: Title: Results Children: Item: Title: Purification of EVs from Conditioned Media from Endometrial Cancer Cells Item: Title: Purification of EVs from Plasma Samples Item: Title: ANXA2 Levels Are Increased in Circulating EVs of EC Patients Item: Title: ANXA2 Levels in Purified EVs from EC Patients Showed Diagnosis Value and Correlated with Tumor Aggressiveness Item: Title: Discussion Item: Title: Materials and Methods Children: Item: Title: Patients Item: Title: EV Production and Isolation from Conditioned Medium Children: Item: Title: Differential Centrifugation Item: Title: ExoGAG Item: Title: Isolation of EVs from Plasma Item: Title: Characterization of EVs Item: Title: ANXA2 and L1CAM Levels in Plasma Samples and Circulating EVs Item: Title: Statistical Analysis Item: Title: Conclusions Item: Title: References Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: OVNEXW+EURM10 FontSubset: true FirstChar: 22 LastChar: 22 FontDescriptor: FontName: OVNEXW+EURM10 Flags: Symbolic FontBBox: -32, -243, 1060, 720 FontFile: true Font: BaseFont: FDYFVT+URWPalladioL-Ital FontSubset: true FirstChar: 2 LastChar: 122 FontDescriptor: FontName: FDYFVT+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: HCUIQB+Pxsy FontSubset: true FirstChar: 0 LastChar: 48 FontDescriptor: FontName: HCUIQB+Pxsy Flags: Symbolic FontBBox: -21, -994, 1292, 912 FontFile: true Font: BaseFont: 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(EVs) are secreted in large amounts into biological fluids of cancer patients. The analysis of EVs cargoes has been associated with patient´s outcome and response to therapy. However, current technologies for EVs isolation are tedious and low cost-efficient for routine clinical implementation. To explore the clinical value of circulating EVs analysis we attempted a proof-of-concept in endometrial cancer (EC) with ExoGAG, an easy to use and highly efficient new technology to enrich EVs. Technical performance was first evaluated using EVs secreted by Hec1A cells. Then, the clinical value of this strategy was questioned by analyzing the levels of two well-known tissue biomarkers in EC, L1 cell adhesion molecule (L1CAM) and Annexin A2 (ANXA2), in EVs purified from plasma in a cohort of 41 EC patients and 20 healthy controls. The results demonstrated the specific content of ANXA2 in the purified EVs fraction, with an accurate sensitivity and specificity for EC diagnosis. Importantly, high ANXA2 levels in circulating EVs were associated with high risk of recurrence and non-endometrioid histology suggesting a potential value as a prognostic biomarker in EC. These results also confirmed ExoGAG technology as a robust technique for the clinical implementation of circulating EVs analyses. 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