Global analyses of genomic and epigenomic influences on gene expression reveal Serpina3n as a major regulator of cardiac gene expression in response to catecholamine challenge during heart failure

Figures

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  Figure 1.. The Hybrid mouse Diversity panel heart failure study a + B) representative circle plots showing the average change in fibrosis (A) or fractional shortening (B) Across the HMDP cohort in response to chronic isoproterenol challenge. C) Schematic for the current study, in which SNP, expression, and methylation information was combined together to identify eQTL, mQTL, and emQTL hotspots.

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  Figure 2.. Representative hotspots for expression and methylation. A) Control eQTL hotspots B) ISO emQTL hotspots C) Delta eQTL hotspots. In all cases, points represent the number of genes/CpGs regulated at a suggestive (p = 4.2 × 10−5) level within a 500kb window. The red line indicates a genome-wide significance threshold.

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  Figure 3.. Serpina3n is the candidate gene for a hotspot on chromosome 12. A) Circular ideogram showing the associations between an eQTL hotspot on chromosome 12 (green bar) with 445 other genes (black bars). Grey lines indicate suggestive association (p < 4.2 × 10−5) while red lines indicate significant associations (4.2 × 10−6). B) Selected enriched terms for genes regulated by this hotspot (full list at Table S5, see also Figure S1). C) Display of 8 of 10 structure-altering mutations in the Serpina3n molecule as rendered by MutationExplorer [50]. Red circles indicate mutation sites, while the yellow circle is the RCL loop active site for the protein. Not displayed are G13R and C20S, the two mutations predicted to be most deleterious, that are located in a disordered region not present in the available structure. D) Correlation of Serpina3n expression with total heart weight across the HMDP panel.

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  Figure 4.. In vitro analysis of Serpina3n knockdown in NRVMs. for all: error bars indicate the SEM, * indicates significance vs control scramble (calculated for other control samples and scramble samples), # indicates significance vs respective control sample (e.G Isoproterenol Serpina3n-a vs. Control Serpina3n-a),! indicates significance vs respective scramble sample (e.g., Isoproterenol Serpina3n-a vs Isoproterenol scramble). Serpina3n-a and Serpina3n-b are the two different siRnas used in this study A) Representative NRVM images B) NRVM cross-sectional areas for cells treated with Isoproterenol, angiotensin II, or phenylephrine and either a scramble siRNA or one of two Serpina3n siRnas. * = P < .05, ** = P < 1 × 10−5, ***=P < 1 × 10−10, ****=P < 1 × 10−20. N = 90 per group, see Table S6 for full details C-M) effects of Serpina3n knockdown on gene expression normalized to scramble control for C) Serpina3n, D+E) control genes Rplp0 and Tnni3, F+G) hypertrophy genes Nppa and Nppb and H-M) putative targets of Serpina3n: acads, Lzic, Wwtr, Pdk4, Agpat9, Lpcat3. For all genes, *=P < 0.05, **=P < 0.005, ***=P < 5 × 10−5. N = 4–15 per group, see Table S7 for full details.