Load-Dependent Effects of Sodium Glucose Co-Transporter Inhibitors on Work in Human Hypertrophic Cardiomyopathy Living Myocardial Slices

Figures

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  Figure 1.. Baseline Work Assessment of hLMS from Non-Failing or HCM Tissue in Glucose or Glucose Supplemented with Ketones. A. Workflow for generation and analysis of hLMS. Freshly explanted human heart tissue from either non-failing donors or from HCM individuals following surgical myectomy are brough to the lab and slices are immediately generated. hLMS are equilibrated in either glucose only containing buffer or glucose and ketone containing buffer. A set of work loops at prespecified preloads and afterloads are generated and processed for analysis. Principal components analysis of all work loop parameter data segmented either by B. heart type or C. fuel. Each dot represents one slice and shaded circles represent 95% confidence ellipses. D. Average work loops ± standard error from the mean (SEM) at all preloads and 50% afterload. Dotted lines represent the end diastolic and end systolic stress strain relationships generated from the full set of 12 work loops. E. Calculated work at 8% strain for each slice at 25, 50, and 75% afterloads. Boxes denote median, 25th and 75th percentile and whiskers represent 1.5 interquartile range (IQR). F. Mixed effect linear model output for work across…

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  Figure 2.. Validation of Human Living Myocardial Slice Work Loops as a Platform to Test Contractility-Modulating Drugs. A. Workflow for drug testing. Following baseline measurement of baseline work loops, LMS is destretched and drug is washed in for 20 minutes. After wash-in, work loop measurements are repeated in the presence of drug. B. PCA of the fold change of work loop parameter data between the drug run and baseline run of work loops. Each dot represents one slice and shaded circles represent 95% confidence ellipses. C. Mixed effect linear model output for work across all preloads at 50% afterload (Mean ± standard error) at baseline (gray) and following drug (pink). D. Average work loops ± standard error from the mean (SEM) at all afterloads and 8% strain. E. Centered work and F. Work strain slope across all loads and heart types at baseline and in response to isoproterenol. G. Centered work and H. Work strain slope across all loads and heart types at baseline and in response to mavacamten. For E-G, bars represent mean ± standard error and values denote p-values between two groups using mixed effects linar regression modeling, pwcompare, and lincom to generate custom contrasts of…

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  Figure 3.. Acute and Direct Effects of SGLTi on Work in Human Living Myocardial Slices. Average work loops ± standard error from the mean (SEM) at all preloads and 50% afterload for A. slices treated with sotagliflozin and B. slices treated with empagliflozin. Dotted lines refer to the end systolic and end diastolic stress strain relationships. C-D. Paired work measurements at baseline and C. in response to sotagliflozin or D. in response to empagliflozin. Boxes denote median, 25th and 75th percentile and whiskers represent 1.5 interquartile range (IQR). E. Mixed effect linear model output at baseline (S refers to eventual sotagliflozin treated slices and E refers to eventual empagliflozin treated slices) and following treatment with SGLTi. Displayed as mean ± standard error. F. Centered work ± standard error and G. work strain slope ± standard error at all afterloads. Values refer to p-values comparing drug to baseline using mixed effects linear regression modeling, pwcompare, and lincom to generate custom contrasts of regression slopes across conditions.