d50ca42499fda32e77bde0bc9609c110 pmc_10380787.pdf 535a761b4cb70b39501360b6e5705d021871b4e5 pmc_10380787.pdf 7c3a4f0077d1234db96ae42ec3f3bd92261f78ed8a465789b18bab612d2d71e7 pmc_10380787.pdf Title: Red Blood Cells and Endothelium Derived Circulating Extracellular Vesicles in Health and Chronic Heart Failure: A Focus on Phosphatidylserine Dynamics in Vesiculation Subject: Circulating extracellular microvesicles (cEVs) are characterised by presenting surface antigens of parental cells. Since their biogenesis involves the translocation of phosphatidylserine (PS) from the inner to the outer leaflet of the plasma membrane, exposed PS has been considered as a recognition hallmark of cEVs. However, not all cEVs externalise PS. In this study, we have phenotypically and quantitatively characterised cEVs by flow cytometry, paying special attention to the proportions of PS in chronic heart failure patients (cHF; n = 119) and a reference non-HF group (n = 21). PS--cEVs were predominantly found in both groups. Parental markers showed differential pattern depending on the PS exposure. Endothelium-derived and connexin 43-rich cEVs were mainly PS--cEVs and significantly increased in cHF. On the contrary, platelet-derived cEVs were mostly PS+ and were increased in the non-HF group. We observed similar levels of PS+- and PS--cEVs in non-HF subjects when analysing immune cell-derived Evs, but there was a subset-specific difference in cHF patients. Indeed, those cEVs carrying CD45+, CD29+, CD11b+, and CD15+ were mainly PS+-cEVs, while those carrying CD14+, CD3+, and CD56+ were mainly PS--cEVs. In conclusion, endothelial and red blood cells are stressed in cHF patients, as detected by a high shedding of cEVs. Despite PS+-cEVs and PS--cEVs representing two distinct cEV populations, their release and potential function as both biomarkers and shuttles for cell communication seem unrelated to their PS content. Keywords: annexin V; connexin 43; endothelial cells; extracellular vesicles; heart failure; leukocytes; microvesicles; phosphatidylserine; platelets; red blood cells Author: Rosa Suades, Alba Vilella-Figuerola, Teresa Padró, Sonia Mirabet and Lina Badimon Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Fri Jul 28 11:29:45 2023 CEST ModDate: Fri Jul 28 11:42:24 2023 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 15 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3777958 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- KPNHET+VnURWPalladioL Type 1 Custom yes yes no 10 0 IVYVDE+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 IRBGMS+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 JFKSCA+URWPalladioL-Ital Type 1 Custom yes yes no 24 0 KXDJCR+CMSY10 Type 1 Builtin yes yes no 28 0 SOZKOC+VnURWPalladioL-Bold Type 1 Custom yes yes no 32 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 50 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 53 0 VVSYYB+EURM10 Type 1 Builtin yes yes no 75 0 QLBIEB+URWPalladioL-BoldItal Type 1 Custom yes yes no 82 0 HYYHEA+CMBSY10 Type 1 Builtin yes yes no 86 0 CNMJDN+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 104 0 CNMJDO+PalatinoLinotype CID TrueType Identity-H yes yes yes 107 0 CNMJDP+PalatinoLinotype TrueType WinAnsi yes yes no 113 0 CNMJEB+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 116 0 CNMKAN+TimesNewRoman CID TrueType Identity-H yes yes yes 119 0 CNMKAO+PalatinoLinotype TrueType MacRoman yes yes no 125 0 CNNDAL+Symbol Type 1C Custom yes yes yes 182 0 CNMJDN+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 187 0 CNMJDO+PalatinoLinotype CID TrueType Identity-H yes yes yes 190 0 CNMJDP+PalatinoLinotype TrueType WinAnsi yes yes no 196 0 CNMJEB+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 199 0 CNMKAN+TimesNewRoman CID TrueType Identity-H yes yes yes 202 0 CNMKAO+PalatinoLinotype TrueType MacRoman yes yes no 208 0 TRNLZC+CMMI10 Type 1 Builtin yes yes no 254 0 CNMKJN+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 269 0 CNMJDN+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 272 0 CNMJDO+PalatinoLinotype CID TrueType Identity-H yes yes yes 275 0 CNMJDP+PalatinoLinotype TrueType WinAnsi yes yes no 281 0 CNMJEB+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 284 0 CNMKAO+PalatinoLinotype TrueType MacRoman yes yes no 287 0 CNMJDN+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 357 0 CNMJDO+PalatinoLinotype CID TrueType Identity-H yes yes yes 360 0 CNMJDP+PalatinoLinotype TrueType WinAnsi yes yes no 366 0 CNMJEB+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 369 0 CNMKAN+TimesNewRoman CID TrueType Identity-H yes yes yes 372 0 CNMKAO+PalatinoLinotype TrueType MacRoman yes yes no 378 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-04-26 02:41:32 CEST RepresentationInformation: pmc_10380787.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-04-24 02:40:28 CEST Size: 3777958 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 665 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: Circulating Extracellular Vesicles with and without Phosphatidylserine Membrane Exposure Destination: subsection.2.1 Item: Title: Differential Patterns between Phosphatidylserine Positive and Negative Circulating Extracellular Vesicles in Non-Heart Failure Reference Group and Patients with Chronic Heart Failure Destination: subsection.2.2 Children: Item: Title: Endothelial-Derived Circulating Extracellular Vesicles Destination: subsubsection.2.2.1 Item: Title: Erythrocyte-Derived Circulating Extracellular Vesicles Destination: subsubsection.2.2.2 Item: Title: Connexin-43-Carrying Circulating Extracellular Vesicles Destination: subsubsection.2.2.3 Item: Title: Platelet and Inflammatory Cell-Derived Circulating Extracellular Vesicles Destination: subsubsection.2.2.4 Item: Title: Discussion Destination: section.3 Item: Title: Conclusions Destination: section.4 Item: Title: Materials and Methods Destination: section.5 Children: Item: Title: Healthy and Diseased Blood Donors Destination: subsection.5.1 Item: Title: Blood Sampling and Circulating Extracellular Vesicle Isolation Destination: subsection.5.2 Item: Title: Flow Cytometric Analysis of Circulating Extracellular Vesicles Destination: subsection.5.3 Item: Title: Statistical Analysis Destination: subsection.5.4 Item: Title: References Destination: section.A. Info: Title: Red Blood Cells and Endothelium Derived Circulating Extracellular Vesicles in Health and Chronic Heart Failure: A Focus on Phosphatidylserine Dynamics in Vesiculation Author: Rosa Suades, Alba Vilella-Figuerola, Teresa Padró, Sonia Mirabet and Lina Badimon Subject: Circulating extracellular microvesicles (cEVs) are characterised by presenting surface antigens of parental cells. Since their biogenesis involves the translocation of phosphatidylserine (PS) from the inner to the outer leaflet of the plasma membrane, exposed PS has been considered as a recognition hallmark of cEVs. However, not all cEVs externalise PS. In this study, we have phenotypically and quantitatively characterised cEVs by flow cytometry, paying special attention to the proportions of PS in chronic heart failure patients (cHF; n = 119) and a reference non-HF group (n = 21). PS--cEVs were predominantly found in both groups. Parental markers showed differential pattern depending on the PS exposure. Endothelium-derived and connexin 43-rich cEVs were mainly PS--cEVs and significantly increased in cHF. On the contrary, platelet-derived cEVs were mostly PS+ and were increased in the non-HF group. We observed similar levels of PS+- and PS--cEVs in non-HF subjects when analysing immune cell-derived Evs, but there was a subset-specific difference in cHF patients. Indeed, those cEVs carrying CD45+, CD29+, CD11b+, and CD15+ were mainly PS+-cEVs, while those carrying CD14+, CD3+, and CD56+ were mainly PS--cEVs. In conclusion, endothelial and red blood cells are stressed in cHF patients, as detected by a high shedding of cEVs. Despite PS+-cEVs and PS--cEVs representing two distinct cEV populations, their release and potential function as both biomarkers and shuttles for cell communication seem unrelated to their PS content. 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