Analysis of Contractile Function of Permeabilized Human Hypertrophic Cardiomyopathy Multicellular Heart Tissue
Analysis of Contractile Function of Permeabilized Human Hypertrophic Cardiomyopathy Multicellular Heart Tissue
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DOI：
10.3389/fphys.2019.00239
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发表时间：
2019-03-28
影响因子：
4
通讯作者：
Friedrich, Felix W.
中科院分区：
文献类型：
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作者：
Kresin, Nico;Stuecker, Sabrina;Friedrich, Felix W.
背景：许多形式的肥厚型心肌病 (HCM) 表现出肌丝 Ca2+ 敏感性增加。这一观察结果主要是在 HCM 小鼠模型、肌丝系统和心肌细胞中进行的。对具有不同 HCM 相关基因突变的患者的多细胞组织的研究很少。我们研究了 HCM 患者多细胞心肌条的 Ca2+ 敏感性。我们还评估了表没食子儿茶素-3-没食子酸酯（EGCg）（一种Ca2+脱敏剂）的使用。方法：从具有单（MYBPC3、MYH7）或双杂合突变（MYBPC3/FLNC、MYH7/LAMP2、MYBPC3/MYH7）的心脏组织中条带分离并透化后，我们进行了收缩性测量+/- EGCg。我们还使用 NanoString 技术通过定制的心力衰竭基因组评估了基因表达。结果：HCM 中的 F-max 往往高于非失败 (NF) 对照条带，单杂合条带中的 F-max 高于双杂合条带中的 F-max。从大多数 HCM 与 NF 条带的趋势以及具有双杂合突变的组织与具有单杂合突变的组织的趋势来看，Ca2+ 敏感性较高。 EGCg 使大多数条带中的肌丝对 Ca2+ 不敏感，并且倾向于在截短突变（比错义突变）或单杂合突变（比双杂合突变）的条带中诱导更明显的转变。基因表达分析显示，与 NF 组织相比，HCM 中 ATP2A2、PPP1R1A 和 FHL2 较低，NPPA、NPPB、COL1A1、CTGF 和 POSTN 标记物水平较高。错义组织中的 NPPA、NPPB、ACTA1、CTGF、COL1A1 和 POSTN 水平高于截短突变组织。结论：我们报告天然多细胞心脏 HCM 条带中肌丝 Ca2+ 敏感性增加，从趋势上看，这种情况在具有双杂合突变的样本中更为明显。 EGCg 可能会产生不同的影响，具体取决于潜在的遗传状态（单杂合与双杂合）和类型（错义与截短）。
Background: Many forms of hypertrophic cardiomyopathy (HCM) show an increased myofilament Ca2+ sensitivity. This observation has been mainly made in HCM mouse models, myofilament systems, and cardiomyocytes. Studies of multicellular tissues from patients with different HCM-associated gene mutations are scarce. We investigated Ca2+ sensitivity in multicellular cardiac muscle strips of HCM patients. We furthermore evaluated the use of epigallocatechin-3-gallate (EGCg), a Ca2+ desensitizer.Methods: After strip isolation from cardiac tissues with single (MYBPC3, MYH7) or double heterozygous mutations (MYBPC3/FLNC, MYH7/LAMP2, MYBPC3/MYH7) and permeabilization, we performed contractility measurements +/- EGCg. We furthermore evaluated gene expression with a customized heart failure gene panel using the NanoString technology.Results: F-max tended to be higher in HCM than in non-failing (NF) control strips and in single than in double heterozygous strips. Ca2+ sensitivity was higher by trend in most HCM vs. NF strips and by trend in tissues with double vs. single heterozygous mutations. EGCg desensitized myofilaments to Ca2+ in most of the strips and tended to induce a more pronounced shift in strips with truncating than missense or single than double heterozygous mutations. Gene expression analysis revealed lower ATP2A2, PPP1R1A, and FHL2 and higher NPPA, NPPB, COL1A1, CTGF, and POSTN marker levels in HCM than in NF tissues. NPPA, NPPB, ACTA1, CTGF, COL1A1, and POSTN levels were higher in tissues with missense than truncating mutations.Conclusion: We report an increased myofilament Ca2+ sensitivity in native multicellular cardiac HCM strips, which by trend was more pronounced in samples with double heterozygous mutations. EGCg could have differential effects depending on the underlying genetic status (single vs. double heterozygous) and type (missense vs. truncating).