Hypertrophic cardiomyopathy β-cardiac myosin mutation (P710R) leads to hypercontractility by disrupting super relaxed state.
Hypertrophic cardiomyopathy β-cardiac myosin mutation (P710R) leads to hypercontractility by disrupting super relaxed state.
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DOI：
10.1073/pnas.2025030118
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发表时间：
2021-06-15
影响因子：
11.1
通讯作者：
Bernstein D
中科院分区：
文献类型：
--
作者：
Vander Roest AS;Liu C;Morck MM;Kooiker KB;Jung G;Song D;Dawood A;Jhingran A;Pardon G;Ranjbarvaziri S;Fajardo G;Zhao M;Campbell KS;Pruitt BL;Spudich JA;Ruppel KM;Bernstein D
心脏病是全世界主要的死亡原因，肥厚性心肌病(HCM)是最常见的遗传性心脏病，每200人中就有1人受到影响。肌球蛋白是导致心脏收缩的运动蛋白，它的突变是引起肥厚性心肌梗死的常见原因，但对肌球蛋白的生物力学有不同的影响。我们证明，与心脏病相关的突变的复杂生物力学效应可以使用多尺度实验和计算建模方法有效地研究和理解。这项工作证实了破坏一个特定的HCM突变的超松弛状态的重要作用，并提供了一种可以扩展的方法，以帮助理解不同突变的疾病机制。肥厚型心肌病是最常见的遗传性心脏病，与1,000多个突变有关，其中许多突变位于β-心肌肌球蛋白(MYH7)。具有不同HCM突变的肌球蛋白的分子研究揭示了对ATPase和从肌动蛋白脱离负荷敏感率的不同影响。很难预测这种不同的分子效应如何结合在一起影响细胞水平的力量，并进一步影响细胞表型。这项研究集中在P710R突变，与其他MYH7突变相比，P710R突变显著降低了体外运动速度和肌动蛋白激活的ATPase。对单个肌球蛋白分子的光学陷阱测量表明，这种突变降低了肌球蛋白马达的步长和肌动蛋白脱落率的负荷敏感度。相反，这种突变破坏了更长的双头肌球蛋白结构中超放松状态的稳定，释放了更多的头来产生力量。带有P710R突变的微图案化人诱导多能干细胞(HiPSC)-心肌细胞CRISPR产生的力(通过牵引力显微镜测量)与同基因对照细胞相比显著增加。通过免疫组织化学染色和电子显微镜观察，P710R突变还导致心肌细胞肥大和细胞骨架重构。抑制ERK或Akt可抑制P710R细胞肥大。最后，我们使用了一个计算模型，该模型集成了测量的分子变化来预测测量的牵引力。这些结果证实了超松弛状态的调节在驱动P710R突变的肥厚性心肌细胞过度收缩中的关键作用，并证明了多尺度方法在揭示疾病关键机制方面的价值。
Heart disease is the leading cause of death worldwide, and hypertrophic cardiomyopathy (HCM) is the most common inherited form of heart disease, affecting over 1 in 200 people. Mutations in myosin, the motor protein responsible for contraction of the heart, are a common cause of HCM but have diverse effects on the biomechanics of the myosin protein. We demonstrate that complex biomechanical effects of mutations associated with heart disease can be effectively studied and understood using a multiscale experimental and computational modeling approach. This work confirms an important role for disruption of the super relaxed state for one particular HCM mutation and provides an approach that can be extended to aid in the understanding of disease mechanisms for different mutations. Hypertrophic cardiomyopathy (HCM) is the most common inherited form of heart disease, associated with over 1,000 mutations, many in β-cardiac myosin (MYH7). Molecular studies of myosin with different HCM mutations have revealed a diversity of effects on ATPase and load-sensitive rate of detachment from actin. It has been difficult to predict how such diverse molecular effects combine to influence forces at the cellular level and further influence cellular phenotypes. This study focused on the P710R mutation that dramatically decreased in vitro motility velocity and actin-activated ATPase, in contrast to other MYH7 mutations. Optical trap measurements of single myosin molecules revealed that this mutation reduced the step size of the myosin motor and the load sensitivity of the actin detachment rate. Conversely, this mutation destabilized the super relaxed state in longer, two-headed myosin constructs, freeing more heads to generate force. Micropatterned human induced pluripotent derived stem cell (hiPSC)–cardiomyocytes CRISPR-edited with the P710R mutation produced significantly increased force (measured by traction force microscopy) compared with isogenic control cells. The P710R mutation also caused cardiomyocyte hypertrophy and cytoskeletal remodeling as measured by immunostaining and electron microscopy. Cellular hypertrophy was prevented in the P710R cells by inhibition of ERK or Akt. Finally, we used a computational model that integrated the measured molecular changes to predict the measured traction forces. These results confirm a key role for regulation of the super relaxed state in driving hypercontractility in HCM with the P710R mutation and demonstrate the value of a multiscale approach in revealing key mechanisms of disease.
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DOI：
10.1073/pnas.1809540115
发表时间：
2018-08-28
影响因子：
11.1
作者：
Anderson, Robert L.;Trivedi, Darshan V.;Spudich, James A.
通讯作者：
Spudich, James A.
影响因子：
8.8
作者：
Adhikari, Arjun S.;Kooiker, Kristina B.;Ruppel, Kathleen M.
通讯作者：
Ruppel, Kathleen M.
影响因子：
3.4
作者：
Campbell, Kenneth S.;Janssen, Paul M. L.;Campbell, Stuart G.
通讯作者：
Campbell, Stuart G.
影响因子：
37.8
作者：
Ho, CY;Sweitzer, NK;Solomon, SD
通讯作者：
Solomon, SD
影响因子：
64.5
作者：
Davis J;Davis LC;Correll RN;Makarewich CA;Schwanekamp JA;Moussavi-Harami F;Wang D;York AJ;Wu H;Houser SR;Seidman CE;Seidman JG;Regnier M;Metzger JM;Wu JC;Molkentin JD
通讯作者：
Molkentin JD