Genomic characterization reveals novel mechanisms underlying the valosin-containing protein-mediated cardiac protection against heart failure

Figures

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  Fig. 1. Overexpression of VCP attenuates pressure-overload induced cardiac pathologic remodeling. The morphological alterations after 5 weeks (5 W) transverse aortic constriction (TAC) in both wild type (WT) and VCP transgenic (TG) mice vs sham. A to B. Alterations of aortic systolic and diastolic blood pressure (ASBP and ADBP) measured by invasive catheter in vivo. C. The representative images of echography. D-E: Left ventricular (LV) posterior wall thickness at end-diastole and systole (LVPWd and LVPWs). F-G. LV end diastolic and systolic dimension (LVEDd and LVEDs). H. LV weight normalized by body weights (LVW/BW). I. The representative images and quantitated cross-sectional areas (CSA) of cardiomyocytes in LV tissues. J. The representative images and quantitated collagen density in LV tissue sections. K. The mRNA levels of ANP and BNP normalized with GAPDH. *p < 0.05, **p < 0.01 vs sham; #p < 0.05, ##p < 0.01 vs corresponding WT mice. N = 5 mice/group.

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  Fig. 2. Overexpression of the VCP prevents pressure-overload induced cardiac dysfunction. The alterations in cardiac function after 5 weeks (5 W) transverse aortic constriction (TAC) in both wild type (WT) and VCP transgenic (TG) mice vs sham. A-C. LV ejection fraction (EF), fractional shortening (FS), and contractile index in all the groups. D-E: LV end-systolic and -diastolic pressure (LVESP and LVEDP). F-G: The maximum or minimum rate of pressure (+dp/dt) in LV isovolumetric contraction. H. Lung weights normalized by body weights. I. The representative images and quantitated apoptosis ratio measured by histology ex vivo in LV tissues via TUNEL. *p < 0.05, **p < 0.01 vs sham; #p < 0.05, ##p < 0.01 vs corresponding WT mice. N = 5 mice/group.

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  Fig. 3. VCP elicits a distinct stress-associated transcriptomic alteration in VCP TG mouse hearts vs WT mice at the end of 5W TAC. A. VCP protein levels normalized to GAPDH. n = 4 mice/group. B–C. Profiles of RNA-seq analysis. Principal component analysis (PCA) (B) and Hierarchical clustering (C) of differentially expressed genes (DEGs) with FDR < 0.05. D. The volcano plot of DEGs between the two groups. Red and green dots represent up- and down-regulated in VCP TG group. E. GO functional analysis of DEGs based on based on the FC > 2 and p < 0.05 between WT and VCP TG mice at the end of 5 W TAC, in terms of the cellular components (CC), molecular function (MF) and biological process (BP) according to p-value. N = 3–4 mice/group for B-E. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

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  Fig. 4. Gene regulations were different between VCP TG and WT mice in response to TAC during the development of heart failure. Hierarchical clustering analysis (A–B) and GO functional analysis of DEGs (C–D) based on the comparison of 5 weeks (5 W) TAC vs sham in either WT (A and C) or VCP TG mice (B and D). CC: the cellular components; MF: the molecular function; and BP: biological process according to p-value. N = 3–4 mice/group.

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  Fig. 5. VCP exhibits a different regulation on the transcription factors by an Ingenuity Pathway Analysis (IPA). A. Il10-ra-mediated downstream genes were inhibited in WT TAC mice as compared to their sham controls. B.Rictor was activated in VCP TG mice upon 5 W TAC, resulting in the repression of the downstream genes related to the oxidative stress. N = 3–4 mice/group.

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  Fig. 6. VCP inhibits ROS-associated gene expression and oxidative damages under the TAC. A-C. The mRNA expression of the genes associated with mitochondrial ROS production including the subunits of mitochondrial complexes I (A), III (B) and IV (C). D-F. The representative images and quantitated indicators of ROS-caused oxidative damages in the LV tissues by the immunofluorescent staining (green) of 8-OHdG (D), 4HNE (E) and MDA (F), combining with the Wheat Germ Agglutinin (WGA) staining (red) for cell membrane and DAPI staining (blue) for nucleic acid. *p < 0.05, **p < 0.01 vs sham, #p < 0.05 vs corresponding WT mice. N = 3–4/groups. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

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  Fig. 7. VCP modulates the alternative splicing of genes in the stressed heart. LeafCutter cluster plots generated by LeafViz for DETXs detected in VCP TG vs WT mice at the end of 5 W TAC showed different modulation of alternative splicing represented by Stra13 (A), Atp5j(B)., Ndrg2 (C) and Atp2a2 (D). N = 3–4 mice/group.