Cell-based delivery of dATP via gap junctions enhances cardiac contractility.
Cell-based delivery of dATP via gap junctions enhances cardiac contractility.
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通过间隙连接进行基于细胞的 dATP 传递可增强心肌收缩力。
DOI：
10.1016/j.yjmcc.2014.04.010
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发表时间：
2014-07
影响因子：
5
通讯作者：
Regnier, Michael
中科院分区：
文献类型：
--
作者：
Lundy, Scott D.;Murphy, Sean A.;Dupras, Sarah K.;Dai, Jin;Murry, Charles E.;Laflamme, Michael A.;Regnier, Michael
关键词：
移植人多能干细胞衍生的心肌细胞（hPSC-CMs）是治疗心肌梗死和逆转心力衰竭的有希望的策略，但迄今为止，大多数研究中的收缩益处仍然有限。我们以前已经表明，核苷酸2-脱氧腺苷三磷酸（dATP）可以取代ATP作为心肌肌球蛋白的能量底物，通过全面过度表达核糖核苷酸还原酶（R1 R2）增加细胞dATP含量可以显着增强心肌收缩力。由于dATP是一种小分子，我们假设它可以通过间隙连接在细胞之间容易地扩散，并增强相邻偶联野生型细胞的收缩性。为了验证这一假设，我们进行了研究，目的是（1）在体外验证间隙连接介导的dATP转移和（2）在体内研究R1 R2过表达的hPSC-CM作为增加心脏功能的新策略的用途。我们首先进行细胞内染料转移研究，使用dATP共轭荧光素，并证明心肌细胞之间的快速间隙连接介导的转移。然后，我们将野生型心肌细胞与过表达R1 R2的心肌细胞或成纤维细胞共培养，发现野生型心肌细胞收缩的程度和速率增加了两倍以上。最后，我们将过表达R1 R2的hPSC-CM移植到健康未受伤的大鼠心脏中，并注意到细胞移植后5天缩短分数从41±4%增加到53±5%。这些发现表明，dATP是一种通过间隙连接在细胞之间传播的正性肌力因子。我们的数据表明，移植产生dATP的hPSC-CM可以显著增加心脏细胞治疗的有效性。
The transplantation of human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) is a promising strategy to treat myocardial infarction and reverse heart failure, but to date the contractile benefit in most studies remains modest. We have previously shown that the nucleotide 2-deoxyadenosine triphosphate (dATP) can substitute for ATP as the energy substrate for cardiac myosin, and increasing cellular dATP content by globally overexpressing ribonucleotide reductase (R1R2) can dramatically enhance cardiac contractility. Because dATP is a small molecule, we hypothesized that it would diffuse readily between cells via gap junctions and enhance the contractility of neighboring coupled wild type cells. To test this hypothesis, we performed studies with the goals of (1) validating gap junction-mediated dATP transfer in vitro and (2) investigating the use of R1R2-overexpressing hPSC-CMs in vivo as a novel strategy to increase cardiac function. We first performed intracellular dye transfer studies using dATP conjugated to fluorescein and demonstrated rapid gap junction-mediated transfer between cardiomyocytes. We then cocultured wild type cardiomyocytes with either cardiomyocytes or fibroblasts overexpressing R1R2 and saw more than a twofold increase in the extent and rate of contraction of wild type cardiomyocytes. Finally, we transplanted hPSC-CMs overexpressing R1R2 into healthy uninjured rat hearts and noted an increase in fractional shortening from 41±4% to 53±5% just five days after cell transplantation. These findings demonstrate that dATP is an inotropic factor that spreads between cells via gap junctions. Our data suggest that transplantation of dATP-producing hPSC-CMs could significantly increase the effectiveness of cardiac cell therapy.
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影响因子：
5.5
作者：
Moreno-Gonzalez, Alicia;Gillis, Todd E.;Regnier, Michael
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