7a70c94a035b6d6d673e6c913136e513 pmc_10855372.pdf 1259077100c97d2a6aa02b48b1a62438ae8e9823 pmc_10855372.pdf ef49d14b2ea82840867448278765ad2ab6a64d7da079e546952e27668bedb4f7 pmc_10855372.pdf Title: TRPV4 Channels Promote Pathological, but Not Physiological, Cardiac Remodeling through the Activation of Calcineurin/NFAT and TRPC6 Subject: TRPV4 channels, which respond to mechanical activation by permeating Ca2+ into the cell, may play a pivotal role in cardiac remodeling during cardiac overload. Our study aimed to investigate TRPV4 involvement in pathological and physiological remodeling through Ca2+-dependent signaling. TRPV4 expression was assessed in heart failure (HF) models, induced by isoproterenol infusion or transverse aortic constriction, and in exercise-induced adaptive remodeling models. The impact of genetic TRPV4 inhibition on HF was studied by echocardiography, histology, gene and protein analysis, arrhythmia inducibility, Ca2+ dynamics, calcineurin (CN) activity, and NFAT nuclear translocation. TRPV4 expression exclusively increased in HF models, strongly correlating with fibrosis. Isoproterenol-administered transgenic TRPV4-/- mice did not exhibit HF features. Cardiac fibroblasts (CFb) from TRPV4+/+ animals, compared to TRPV4-/-, displayed significant TRPV4 overexpression, elevated Ca2+ influx, and enhanced CN/NFATc3 pathway activation. TRPC6 expression paralleled that of TRPV4 in all models, with no increase in TRPV4-/- mice. In cultured CFb, the activation of TRPV4 by GSK1016790A increased TRPC6 expression, which led to enhanced CN/NFATc3 activation through synergistic action of both channels. In conclusion, TRPV4 channels contribute to pathological remodeling by promoting fibrosis and inducing TRPC6 upregulation through the activation of Ca2+-dependent CN/NFATc3 signaling. These results pose TRPV4 as a primary mediator of the pathological response. Keywords: TRPV4; TRPC6; calcium; mechanotransduction; mechanoreceptors; TRP; pathological remodeling; heart failure; physiological remodeling; exercise Author: Laia Yáñez-Bisbe, Mar Moya, Antonio Rodríguez-Sinovas, Marisol Ruiz-Meana, Javier Inserte, Marta Tajes, Montserrat Batlle, Eduard Guasch, Aleksandra Mas-Stachurska, Elisabet Miró, Nuria Rivas, Ignacio Ferreira González, Anna Garcia-Elias and Begoña Benito Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Fri Jan 26 10:49:49 2024 CET ModDate: Fri Jan 26 11:02:59 2024 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 20 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3942655 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- PRJKGQ+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 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PDFMetadata: Objects: 520 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: Differential Expression of Cardiac TRPV4 Channels in Physiological and Pathological Remodeling Destination: subsection.2.1 Item: Title: Time-Course Changes in TRPV4 Expression throughout the Development of Pathological Cardiac Remodeling Destination: subsection.2.2 Item: Title: Effects of TRPV4 Deletion on the Development of Pathological Cardiac Remodeling Destination: subsection.2.3 Item: Title: TRPV4 Overexpression in Pathological Remodeling Induces Enhanced Ca2+ Influx in Cardiac Fibroblasts Destination: subsection.2.4 Item: Title: TRPV4 Activation in Pathological Remodeling Mediates Fibrosis though Activation of the CN/NFATc3 Pathway Destination: subsection.2.5 Item: Title: The Interplay between TRPV4 and TRPC6 in the Promotion of Pathological Cardiac Remodeling Destination: subsection.2.6 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Animals Destination: subsection.4.1 Item: Title: In Vivo Models of Adverse and Adaptive Cardiac Remodeling Destination: subsection.4.2 Item: Title: Echocardiography Measurements Destination: subsection.4.3 Item: Title: Histology Destination: subsection.4.4 Item: Title: Langendorff-Perfused Hearts Destination: subsection.4.5 Item: Title: Assessment of Arrhythmias Destination: subsection.4.6 Item: Title: Fibroblast Culture Destination: subsection.4.7 Item: Title: qPCR Analyses Destination: subsection.4.8 Item: Title: Western Blotting Destination: subsection.4.9 Item: Title: Fluorescence Ca2+ Imaging Destination: subsection.4.10 Item: Title: Calcineurin Activity Assay Destination: subsection.4.11 Item: Title: NFAT Nuclear Translocation Destination: subsection.4.12 Item: Title: Proximity Ligation Assay Destination: subsection.4.13 Item: Title: Statistical Analysis Destination: subsection.4.14 Item: Title: References Destination: appendix.A. Info: Title: TRPV4 Channels Promote Pathological, but Not Physiological, Cardiac Remodeling through the Activation of Calcineurin/NFAT and TRPC6 Author: Laia Yáñez-Bisbe, Mar Moya, Antonio Rodríguez-Sinovas, Marisol Ruiz-Meana, Javier Inserte, Marta Tajes, Montserrat Batlle, Eduard Guasch, Aleksandra Mas-Stachurska, Elisabet Miró, Nuria Rivas, Ignacio Ferreira González, Anna Garcia-Elias and Begoña Benito Subject: TRPV4 channels, which respond to mechanical activation by permeating Ca2+ into the cell, may play a pivotal role in cardiac remodeling during cardiac overload. Our study aimed to investigate TRPV4 involvement in pathological and physiological remodeling through Ca2+-dependent signaling. TRPV4 expression was assessed in heart failure (HF) models, induced by isoproterenol infusion or transverse aortic constriction, and in exercise-induced adaptive remodeling models. The impact of genetic TRPV4 inhibition on HF was studied by echocardiography, histology, gene and protein analysis, arrhythmia inducibility, Ca2+ dynamics, calcineurin (CN) activity, and NFAT nuclear translocation. TRPV4 expression exclusively increased in HF models, strongly correlating with fibrosis. Isoproterenol-administered transgenic TRPV4-/- mice did not exhibit HF features. Cardiac fibroblasts (CFb) from TRPV4+/+ animals, compared to TRPV4-/-, displayed significant TRPV4 overexpression, elevated Ca2+ influx, and enhanced CN/NFATc3 pathway activation. TRPC6 expression paralleled that of TRPV4 in all models, with no increase in TRPV4-/- mice. In cultured CFb, the activation of TRPV4 by GSK1016790A increased TRPC6 expression, which led to enhanced CN/NFATc3 activation through synergistic action of both channels. In conclusion, TRPV4 channels contribute to pathological remodeling by promoting fibrosis and inducing TRPC6 upregulation through the activation of Ca2+-dependent CN/NFATc3 signaling. These results pose TRPV4 as a primary mediator of the pathological response. Keywords: TRPV4; TRPC6; calcium; mechanotransduction; mechanoreceptors; TRP; pathological remodeling; heart failure; physiological remodeling; exercise Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Fri Jan 26 10:49:49 CET 2024 ModDate: Fri Jan 26 11:02:59 CET 2024 ID: 0xedc460710fba06b5e48ac45524a0aff6, 0xedc460710fba06b5e48ac45524a0aff6 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: MOCBVF+VnURWPalladioL FontSubset: true FirstChar: 129 LastChar: 243 FontDescriptor: FontName: MOCBVF+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: PZNAJG+URWPalladioL-Ital FontSubset: true FirstChar: 3 LastChar: 121 FontDescriptor: FontName: PZNAJG+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: GXYDYO+URWPalladioL-Bold FontSubset: true FirstChar: 3 LastChar: 241 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