Molecular Signature of HFpEF

Figures

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  Figure 1. 1-Month Aortic Banding Reveals a Cardiac RNAseq Signature of Transcriptional and Post-Transcriptional Regulation, Mitochondrial Dysfunction, and Inflammation Differential expression and gene enrichment analysis in hearts subjected to RNAseq 1-month post-banding were compared with sham controls. (A) Volcano plot of differentially expressed RNA transcripts indicating the number of genes increased and decreased in expression (fold change ≥1.5; FDR ≤ 0.1); (B) heatmap of the 99 genes found to be differentially expressed. Gene ontology (GO) analysis revealing the (C) biological processes and (D) molecular functions of those genes found to be differentially expressed. n = 3 sham and 5 banded. FDR = false discovery rate.

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  Figure 2. Cardio-Metabolic Remodeling Is a Characteristic Feature of Early HFpEF Development Metabolic phenotyping of hearts subjected to 1-month aortic banding compared with sham controls: (A) Volcano plot analysis showing metabolites with a fold change (FC) ≥1.25-fold; P ≤ 0.1; (B) heatmap of the top 50 significantly different metabolites; (C) metabolic network visualization of metabolites in 1-month banded hearts as compared with sham control hearts. Colored circles indicate a statistically significant difference with the size of the circle indicative of the degree of change: 1. gamma-glutamyl amino acids; 2. inosine purines; 3. cytidine pyrimidines; 4. adenine purines; 5. uracil pyrimidines; 6. long-chain polyunsaturated fatty acids; 7. poly-, un-, saturated fatty acid species; 8. phospholipids; 9. mono- and dicarboxylic fatty acids; 10. monoacylglycerols; 11. TCA cycle; 12. vitamin B6 metabolism; 13. ascorbate metabolism; 14. Met, Cys, SAM metabolism; 15. lysine metabolism; 16. Leu, Ile, Val metabolism; 17. His metabolism; 18. Ala, Asp metabolism; 19. Gly, Ser, Thr metabolism; 20. Trp metabolism; 21. glycolytic intermediates; 22. amino sugars; (D) enrichment overview of the metabolic…

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  Figure 3. Correlations Between the Cardio-Metabolic Profile and Mitochondrial Function Highlight an Energetic and Functional Decline During Early Cardiac Remodeling (A and B) Interrogation of mitochondrial function following 1-month of aortic banding: state 3 (substrate-mediated), state 4 (oligomycin inhibition of complex V), and FCCP-mediated (maximal respiratory capacity) oxygen consumption rates (OCRs) in isolated cardiac mitochondria. Quantification of state 3, state 4, and FCCP-mediated rates for (C) complex I (pyruvate + malate)-specific substrates and (D) complex II (succinate)-specific substrates + rotenone (complex I inhibitor); (E) quantification of respiratory control ratios (state 3/state 4), providing an index of oxygen consumption to ATP-production coupling; Metabolite correlations with (F) complex I – state 3 and (G) FCCP mediated rates; metabolite correlations with (H) complex II–state 3 and (I) respiratory control ratio (RCR). Metabolite correlations with (J) cardiac fibrosis. The raw metabolomic data from each individual cat were used to make the correlations to the functional parameters indicated in each graph. All correlations have a P ≤ 0.1. n = 3 sham and 5 banded. ∗P…

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  Figure 4. Changes in Metabolism, Cell Signaling, and Inflammation Define the Cardiac Transcriptomic Signature Following the Progression to HFpEF Metabolomic and RNAseq analyses of hearts 4 months post-banding compared with age-matched shams. (A) Volcano plot of differentially expressed RNA transcripts indicating the number of genes increased and decreased in expression (fold change [FC] ≥1.5; FDR ≤ 0.1); (B) heatmap of the top 100 genes found to be differentially expressed. Gene ontology (GO) analysis revealing the (C) biological processes and (D) molecular functions of those genes found to be differentially expressed. n = 5 sham and 5 banded. FDR = false discovery rate; HFpEF = heart failure with preserved ejection fraction.

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  Figure 5. Metabolic Analysis of the Heart 4 Months Post-Banding Reveals a Sustained Remodeling of Amino Acid Metabolism and a Greater Reliance on Aerobic Glycolysis for Energy Provision Metabolic phenotyping of hearts 4 months post-aortic banding compared with age-matched shams: (A) Volcano plot analysis showing metabolites with a fold change (FC) ≥1.25-fold; P ≤ 0.1; (B) heatmap of the top 50 significantly different metabolites; (C) metabolic network visualization of metabolites in 4-month banded hearts as compared with sham control hearts. Colored circles indicate a statistically significant difference with the size of the circle indicative of the degree of change: 1. gamma-glutamyl amino acids; 2. inosine purines; 3. cytidine pyrimidines; 4. adenine purines; 5. uracil pyrimidines; 6. long-chain polyunsaturated fatty acids; 7. poly-, un-, saturated fatty acid species; 8. phospholipids; 9. mono- and dicarboxylic fatty acids; 10. monoacylglycerols; 11. TCA cycle; 12. vitamin B6 metabolism; 13. ascorbate metabolism; 14. Met, Cys, SAM metabolism; 15. lysine metabolism; 16. Leu, Ile, Val metabolism; 17. His metabolism; 18. Ala, Asp metabolism; 19. Gly, Ser, Thr metabolism; 20. Trp metabolism;…

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  Figure 6. Skeletal Muscle-omics Profiling Indicates an Energetic Deficit in Early HFpEF Progression Transcriptomic and metabolomic analysis of skeletal muscle (Skm) following 1-month aortic banding compared with sham controls. (A) Volcano plot of differentially expressed RNA transcripts indicating the number of genes increased and decreased in expression (FC ≥1.5; FDR ≤0.1); (B) heatmap of differentially expressed genes; (C) volcano plot analysis showing metabolites with a FC ≥1.25-fold; P ≤ 0.1; (D) metabolite heatmap profile. n = 3 sham 1-month and 5 banded 1-month. Abbreviations as in Figures 2 and 3.

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  Figure 7. Remodeling of Skeletal Muscle 4 Months Post-Banding Is Defined by a Defective Oxidative Metabolism Transcriptomic and metabolomic analysis of skeletal muscle (Skm) following 4-month aortic banding compared with sham controls. (A) Volcano plot of differentially expressed RNA transcripts indicating the number of genes increased and decreased in expression (FC ≥ 1.5; FDR ≤0.1). (B) heatmap of differentially expressed genes (FC ≥1.5; P ≤ 0.1; total of 163 genes); (C) gene ontology (GO) analysis of the biological processes of the differentially expressed genes; (D) volcano plot analysis showing metabolites with a FC ≥1.25-fold; P ≤ 0.1; (E) metabolite heatmap profile. n = 5 sham 4-month and 5 banded 4-month. Abbreviations as in Figures 2 and 3.

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  Figure 8. Mitochondrial Dysfunction Persists in the Skeletal Muscle During HFpEF Progression (A and D) Interrogation of mitochondrial function following 4 months of aortic banding: state 3 (substrate-mediated), state 4 (oligomycin inhibition of complex V), and FCCP-mediated (maximal respiratory capacity) oxygen consumption rates in isolated cardiac mitochondria. Quantification of state 3, state 4, and FCCP-mediated rates for (B) complex I (pyruvate + malate)-specific substrates and (E) complex II (succinate)-specific substrates + rotenone (complex I inhibitor); (C and F) Quantification of respiratory control ratios (state 3/state 4), providing an index of oxygen consumption to ATP-production coupling. n = 4 sham 4-month and 4 banded 4-month. Abbreviations as in Figures 2 and 3.