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Mechanical stretch induced transcriptomic profiles in cardiac myocytes II

ABSTRACT: The effect of cyclic mecanical stretch on cardiac gene expression was studied in neonatal rat ventricular myocytes (NRVMs).

ORGANISM(S): Rattus norvegicus

PROVIDER: GSE107551 | GEO | 2018/03/17

REPOSITORIES: GEO

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Mechanical stretch induced transcriptomic profiles in cardiac myocytes [24-48 hours]

Project description:The effect of cyclic mecanical stretch on cardiac microRNA expression was studied in neonatal rat ventricular myocytes (NRVMs).

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Project description:The effect of cyclic mecanical stretch on cardiac microRNA expression was studied in neonatal rat ventricular myocytes (NRVMs).

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Mechanical stretch induced transcriptomic profiles in cardiac myocytes II

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Mechanical stretch induced transcriptomic profiles in cardiac myocytes [1-12 hours]

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Project description:Myocardial damage caused for example by cardiac ischemia leads to ventricular volume overload resulting in increased stretch of the remaining myocardium. In adult mammals, these changes trigger an adaptive cardiomyocyte hypertrophic response which, if the damage is extensive, will ultimately lead to pathological hypertrophy and heart failure. Conversely, in response to extensive myocardial damage, cardiomyocytes in the adult zebrafish heart and neonatal mice proliferate and completely regenerate the damaged myocardium. We therefore hypothesized that in adult zebrafish, changes in mechanical loading due to myocardial damage may act as a trigger to induce cardiac regeneration. Based, on this notion we sought to identify mechanosensors which could be involved in detecting changes in mechanical loading and triggering regeneration. Here we show using a combination of knockout animals, RNAseq and in vitro assays that the mechanosensitive ion channel Trpc6a is required by cardiomyocytes for successful cardiac regeneration in adult zebrafish. Furthermore, using a cyclic cell stretch assay, we have determined that Trpc6a induces the expression of components of the AP1 transcription complex in response to mechanical stretch. Our data highlights how changes in mechanical forces due to myocardial damage can be detected by mechanosensors which in turn can trigger cardiac regeneration.

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Project description:The goal of this study was to examine the effect of the major axis of biaxial mechanical stretch on cardiac myocyte gene expression and to identify the signaling pathways and transcription factors regulating these changes. Neonatal cardiac myocytes were cultured on a micropatterned substrate, and the primary stretch axis was applied either parallel or transverse to the myofibril direction. RNA sequencing was conducted to study whole genomic expression changes after acute cardiac myocyte stretch. The results showed a more robust gene response to longitudinal than to transverse stretch. After 30 minutes of stretch, 53 and 168 genes were considered differentially expressed (DE) from transverse and longitudinal stretch, respectively. After 4 hours, the number of DE genes increased to 795 in longitudinal stretch while it decreased to 35 in transverse stretch. Gene ontology term (GO) analysis indicated enrichment of TF activity and protein kinase activity by both stretch axes; whereas longitudinal but not transverse stretch caused expression of genes involved in sarcomere organization and cytoskeletal protein binding.

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Project description:In this study, we performed two-dimensional liquid chromatography coupled to tandem mass spectrometry (2D-LC-MS/MS) to characterize the global transcriptome and proteome changes in Tbx18-iPMs compared to control, GFP-overexpressing neonatal rat ventricular myocytes (NRVMs). 67% of protein abundances changed significantly (4,475 of 6,661 proteins; p<0.05) at a false discovery rate of 1.6%. Downregulation was broad among metabolic pathways, such as TCA cycle, oxidative phosphorylation, glycolysis and fatty acid oxidation. Ion channels associated with ventricular excitation-contraction coupling (sodium, calcium, and potassium channels, Connexin 43, among others) were likewise downregulated. Pacemaker phenotype was characterized by upregulation of the pacemaker current channel (HCN4) but also mechnosensitive Piezo1, Trp channels Trpp2 (PKD2) and TrpM7, and Connexin45. Consistent with emergent pacemaker morphology, there was broad upregulation of cytoskeletal proteins and their small GTPase regulators. Extracellular matrix and growth factor/cytokine/interferon genes were also dynamically upregulated, indicating that Tbx18-iPMs are primed for sinoatrial node integration.

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