e7b90e74c50bbf73301ab74fe175eb0b pmc_34638717.pdf 6fcd4ed7f194e5dae2266ab1be9e2961c30ea62d pmc_34638717.pdf 343ea1bca5c8410e364681312a530680c08e2572685f6a96eed3847401dffce4 pmc_34638717.pdf Title: Atrial Fibrillation in Heart Failure Is Associated with High Levels of Circulating microRNA-199a-5p and 22–5p and a Defective Regulation of Intracellular Calcium and Cell-to-Cell Communication Subject: MicroRNAs (miRNAs) participate in atrial remodeling and atrial fibrillation (AF) promotion. We determined the circulating miRNA profile in patients with AF and heart failure with reduced ejection fraction (HFrEF), and its potential role in promoting the arrhythmia. In plasma of 98 patients with HFrEF (49 with AF and 49 in sinus rhythm, SR), differential miRNA expression was determined by high-throughput microarray analysis followed by replication of selected candidates. Validated miRNAs were determined in human atrial samples, and potential arrhythmogenic mechanisms studied in HL-1 cells. Circulating miR-199a-5p and miR-22-5p were significantly increased in HFrEF patients with AF versus those with HFrEF in SR. Both miRNAs, but particularly miR-199a-5p, were increased in atrial samples of patients with AF. Overexpression of both miRNAs in HL-1 cells resulted in decreased protein levels of L-type Ca2+ channel, NCX and connexin-40, leading to lower basal intracellular Ca2+ levels, fewer inward currents, a moderate reduction in Ca2+ buffering post-caffeine exposure, and a deficient cell-to-cell communication. In conclusion, circulating miR-199a-5p and miR-22-5p are higher in HFrEF patients with AF, with similar findings in human atrial samples of AF patients. Cells exposed to both miRNAs exhibited altered Ca2+ handling and defective cell-to-cell communication, both findings being potential arrhythmogenic mechanisms.40pt Keywords: heart failure; atrial fibrillation; atrial remodeling; microRNA; biomarkers; HL-1 cells; calcium regulation; L-type calcium channels; connexin 40; NCX1 Author: Anna Garcia-Elias, Marta Tajes, Laia Yañez-Bisbe, Cristina Enjuanes, Josep Comín-Colet, Selma A. Serra, José M. Fernández-Fernández, Kathryn W. Aguilar-Agon, Svetlana Reilly, Julio Martí-Almor and Begoña Benito Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Sep 27 09:54:22 2021 CEST ModDate: Mon Sep 27 10:38:16 2021 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 17 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3018063 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- AWHDWW+VnURWPalladioL Type 1 Custom yes yes no 10 0 VCADXR+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 HSMLFN+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 MSJCAM+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 IOJCNK+VnURWPalladioL-Bold Type 1 Custom yes yes no 28 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 36 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 39 0 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1.28.0, 2023-05-18) Date: 2025-11-23 05:27:23 CET RepresentationInformation: pmc_34638717.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-03-31 12:05:51 CEST Size: 3018063 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 479 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: HFrEF Patients with AF Have Higher Plasmatic Levels of miRNA-199a-5p and miRNA-22-5p Destination: subsection.2.1 Item: Title: MiR-199a-5p and 22 5p Expression Levels Are Higher in Atrial Samples of AF Patients Destination: subsection.2.2 Item: Title: Overexpression of miR-199a-5p and miR-22-5p Is Associated with Reduced Expression of Proteins Involved in Ca2+ Homeostasis and Cell-to-Cell Communication Destination: subsection.2.3 Item: Title: Overexpression of miR-199a-5p and miR-22-5p Is Associated with Reduced Intracellular Ca2+ Levels and a Decrease in Barium Currents (IBa) Destination: subsection.2.4 Item: Title: Cells Transfected with miR-199a-5p and miR-22-5p Have Higher Ca2+ Levels in the Sarcoplasmic Reticulum and a Slower Caffeine-Induced Ca2+ Decay Destination: subsection.2.5 Item: Title: Overexpression of miR-199a-5p and miR-22-5p Is Associated with a Reduction in Cx40 and Impaired Cell-to-Cell Communication Destination: subsection.2.6 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Human Plasma Study Destination: subsection.4.1 Children: Item: Title: Study Population Destination: subsubsection.4.1.1 Item: Title: Experimental Design Destination: subsubsection.4.1.2 Item: Title: MicroRNA Extraction and Profiling Destination: subsection.4.2 Item: Title: Human Atrial Samples Destination: section.5 Children: Item: Title: Cardiac Tissue Collection Destination: subsection.5.1 Item: Title: MicroRNA Profiling Destination: subsection.5.2 Item: Title: Molecular Studies Destination: section.6 Children: Item: Title: Cell Cultures and Transfection Destination: subsection.6.1 Children: Item: Title: qPCR and Western Blot Analyses Destination: subsubsection.6.1.1 Item: Title: Recordings of Intracellular Ca2+ and Patch Clamp Studies Destination: subsubsection.6.1.2 Item: Title: Gap-Junction Monitoring Destination: subsubsection.6.1.3 Item: Title: Statistical Analyses Destination: subsection.6.2 Item: Title: Conclusions Destination: section.7 Item: Title: References Destination: section.7 Info: Title: Atrial Fibrillation in Heart Failure Is Associated with High Levels of Circulating microRNA-199a-5p and 22 5p and a Defective Regulation of Intracellular Calcium and Cell-to-Cell Communication Author: Anna Garcia-Elias, Marta Tajes, Laia Yañez-Bisbe, Cristina Enjuanes, Josep Comín-Colet, Selma A. Serra, José M. Fernández-Fernández, Kathryn W. Aguilar-Agon, Svetlana Reilly, Julio Martí-Almor and Begoña Benito Subject: MicroRNAs (miRNAs) participate in atrial remodeling and atrial fibrillation (AF) promotion. We determined the circulating miRNA profile in patients with AF and heart failure with reduced ejection fraction (HFrEF), and its potential role in promoting the arrhythmia. In plasma of 98 patients with HFrEF (49 with AF and 49 in sinus rhythm, SR), differential miRNA expression was determined by high-throughput microarray analysis followed by replication of selected candidates. Validated miRNAs were determined in human atrial samples, and potential arrhythmogenic mechanisms studied in HL-1 cells. Circulating miR-199a-5p and miR-22-5p were significantly increased in HFrEF patients with AF versus those with HFrEF in SR. Both miRNAs, but particularly miR-199a-5p, were increased in atrial samples of patients with AF. Overexpression of both miRNAs in HL-1 cells resulted in decreased protein levels of L-type Ca2+ channel, NCX and connexin-40, leading to lower basal intracellular Ca2+ levels, fewer inward currents, a moderate reduction in Ca2+ buffering post-caffeine exposure, and a deficient cell-to-cell communication. In conclusion, circulating miR-199a-5p and miR-22-5p are higher in HFrEF patients with AF, with similar findings in human atrial samples of AF patients. Cells exposed to both miRNAs exhibited altered Ca2+ handling and defective cell-to-cell communication, both findings being potential arrhythmogenic mechanisms.40pt Keywords: heart failure; atrial fibrillation; atrial remodeling; microRNA; biomarkers; HL-1 cells; calcium regulation; L-type calcium channels; connexin 40; NCX1 Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Sep 27 09:54:22 CEST 2021 ModDate: Mon Sep 27 10:38:16 CEST 2021 ID: 0xf91aca169b4dd020fce4b248f0406a86, 0xf91aca169b4dd020fce4b248f0406a86 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: HSMLFN+URWPalladioL-Bold FontSubset: true FirstChar: 3 LastChar: 241 FontDescriptor: FontName: HSMLFN+URWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: QLBIEB+URWPalladioL-BoldItal FontSubset: true FirstChar: 110 LastChar: 112 FontDescriptor: FontName: QLBIEB+URWPalladioL-BoldItal Flags: Symbolic FontBBox: 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