DNA Tension Probes Show that Cardiomyocyte Maturation Is Sensitive to the Piconewton Traction Forces Transmitted by Integrins.
DNA Tension Probes Show that Cardiomyocyte Maturation Is Sensitive to the Piconewton Traction Forces Transmitted by Integrins.
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DOI：
10.1021/acsnano.1c04303
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发表时间：
2022-03
期刊：
影响因子：
17.1
通讯作者：
S. A. Rashid;Aaron T. Blanchard;J. D. Combs;Natasha Fernandez;Yixiao Dong;H. Cho;K. Salaita
中科院分区：
文献类型：
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作者：
S. A. Rashid;Aaron T. Blanchard;J. D. Combs;Natasha Fernandez;Yixiao Dong;H. Cho;K. Salaita
心肌细胞（CMC）是构成心脏的单位细胞。CMC经历不同的分化和成熟途径阶段，以完全成熟为搏动细胞。这些细胞可以通过影响成熟途径的受体如整合素来感知和响应机械信号。例如，细胞牵引力对于功能性CMC的分化和发育是重要的，因为在不同基质硬度上培养的CMC功能不同。这一领域的大多数工作都集中在理解大量细胞外基质硬度在介导CMC功能命运中的作用。鉴于刚度传感机制是由个体整联蛋白受体介导的，该领域的一个重要问题涉及可以触发CMC功能成熟的整联蛋白皮牛顿（pN）力的具体大小。为了解决这一知识缺口，我们使用了在特定力阈值（12 pN、156 pN和160 pN）下断裂的DNA粘附系链来测试将峰值整合素张力加帽至特定幅度是否会影响CMC功能。我们发现，具有更大的力耐受性的粘附栓系导致功能成熟的CMC，如通过形态学、抽搐频率、瞬时钙通量测量和蛋白质表达（F-肌动蛋白、黏着斑蛋白、α-辅肌动蛋白、雅普和SERCA 2a）所确定的。此外，肌节辅肌动蛋白对齐和多核化显着增强的机械耐受性的整联蛋白栓。总之，结果表明CMC利用定义的pN整联蛋白力来影响早期发育。这项研究是一个重要的一步，生物物理特性的贡献pN部队在早期阶段的心脏分化。
Cardiac muscle cells (CMCs) are the unit cells that comprise the heart. CMCs go through different stages of differentiation and maturation pathways to fully mature into beating cells. These cells can sense and respond to mechanical cues through receptors such as integrins which influence maturation pathways. For example, cell traction forces are important for the differentiation and development of functional CMCs, as CMCs cultured on varying substrate stiffness function differently. Most work in this area has focused on understanding the role of bulk extracellular matrix stiffness in mediating the functional fate of CMCs. Given that stiffness sensing mechanisms are mediated by individual integrin receptors, an important question in this area pertains to the specific magnitude of integrin piconewton (pN) forces that can trigger CMC functional maturation. To address this knowledge gap, we used DNA adhesion tethers that rupture at specific thresholds of force (∼12, ∼56, and ∼160 pN) to test whether capping peak integrin tension to specific magnitudes affects CMC function. We show that adhesion tethers with greater force tolerance lead to functionally mature CMCs as determined by morphology, twitching frequency, transient calcium flux measurements, and protein expression (F-actin, vinculin, α-actinin, YAP, and SERCA2a). Additionally, sarcomeric actinin alignment and multinucleation were significantly enhanced as the mechanical tolerance of integrin tethers was increased. Taken together, the results show that CMCs harness defined pN integrin forces to influence early stage development. This study represents an important step toward biophysical characterization of the contribution of pN forces in early stage cardiac differentiation.