Analysis of Contractile Function of Permeabilized Human Hypertrophic Cardiomyopathy Multicellular Heart Tissue

Figures

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  Figure 1. Force development of permeabilized cardiac muscle strips isolated from seven HCM patients with either single heterozygous (MYBPC3, MYH7) or double heterozygous mutations (MYH7/LAMP2, MYBPC3/MYH7, MYBPC3/FLNC). Maximal force development (Fmax) related to cross-sectional area in non-failing control vs. HCM strips (A), grouped in missense or truncating (B), single or double heterozygous mutations (C), and classified according to a pure MYBPC3 or MYH7 genotype (D). Values are related to non-failing control (NF) strips. Data are expressed as mean ± SEM. **p < 0.01 and ***p < 0.001 vs. NF, mixed-effect model analysis followed by Dunnett multiple comparison post-test; for the different subgroup analyses, only qualitative comparisons were performed. n = number of strips; n/N = number of strips/number of patients as depicted in the bars.

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  Figure 2. Force-Ca2+ relationship of permeabilized cardiac muscle strips isolated from seven HCM patients with either single heterozygous (MYBPC3, MYH7) or double heterozygous mutations (MYH7/LAMP2, MYBPC3/MYH7, MYBPC3/FLNC). (A) Force-Ca2+ relationship curves in human strips, Hill slope set to 3. (B) pCa50 representing the negative logarithm of the Ca2+ concentration needed for half-maximal activation. (C–E) pCa50 in strips grouped according to mutation type (missense or truncating), to genetic status (single or double heterozygous mutations), and to a pure MYBPC3 or MYH7 genotype. (F) nHill coefficient (Hill slope). Values are related to non-failing control (NF) strips. Data are expressed as mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001 vs. NF, mixed-effect model analysis followed by Dunnett multiple comparison post-test; for the different subgroup analyses, only qualitative comparisons were performed; concentration response curves were fitted to the data points and curve comparison was done by using extra sum-of-squares F-test. n = number of strips; n/N = number of strips/number of patients as depicted in the bars.

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  Figure 3. Effect of 30 μM EGCg on pCa50 values in permeabilized cardiac muscle strips isolated from seven HCM patients with either single heterozygous (MYBPC3, MYH7) or double heterozygous mutations (MYH7/LAMP2, MYBPC3/MYH7, MYBPC3/FLNC). (A) Delta of pCa50 ± EGCg (30 μM) in non-failing control vs. HCM permeabilized cardiac muscle strips grouped according to mutation type (missense or truncating, (B), to genetic status (single or double heterozygous mutations, (C), and to a pure MYBPC3, MYH7, or ACTN2 genotype (D). Values are related to baseline. Data are expressed as mean ± SEM. ***p < 0.001 vs. baseline (without EGCg), paired Student’s t-test; for the different subgroup analyses, only qualitative comparisons were performed. n = number of strips; n/N = number of strips/number of patients as depicted in the bars.

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  Figure 4. Evaluation of expression of human cardiac genes regulated in heart failure in NF and HCM tissues. Normalized expression levels of selected genes determined with the NanoString nCounter Elements technology comparing gene expression modulating hypertrophy, fibrosis, calcium handling, and apoptosis in NF and HCM (A), (threshold <0.8- or > 1.25-fold change to NF). Control group consisted of RNA isolated from eight different NF control hearts. Gene expression related to (B) mutation type (missense or truncating) or (C) genetic status (single or double heterozygous mutation). Since for patient #7 (Table 1) obtained results repeatedly failed, quality control check data were not included in the analysis.