Identification of chemicals inducing cardiomyocyte proliferation in developmental stage-specific manner with pluripotent stem cells.
Identification of chemicals inducing cardiomyocyte proliferation in developmental stage-specific manner with pluripotent stem cells.
复制标题
DOI：
10.1161/circgenetics.113.000330
复制
发表时间：
2013-12
期刊：
影响因子：
--
通讯作者：
Yamashita JK
中科院分区：
文献类型：
--
作者：
Uosaki H;Magadum A;Seo K;Fukushima H;Takeuchi A;Nakagawa Y;Moyes KW;Narazaki G;Kuwahara K;Laflamme M;Matsuoka S;Nakatsuji N;Nakao K;Kwon C;Kass DA;Engel FB;Yamashita JK
出生后成熟后，心肌细胞的增殖受到高度限制，限制了心脏再生。阐明心肌细胞增殖和生长停止的调控机制是必要的。化学生物学是有效的解剖各种细胞事件的分子机制，并经常提供治疗潜力。我们一直在研究多能干细胞（PSCs）的心血管分化。干细胞与化学生物学的结合为研究心肌细胞增殖的分子机制和调控提供了新的途径。为了确定调节心肌细胞增殖的化学物质，我们基于小鼠psc来源的心肌细胞的增殖，对一个定义的化学文库进行了筛选，并确定了4个化合物组-糖原合成酶激酶3 （GSK3）抑制剂，p38丝裂原活化蛋白激酶（MAPK）和Ca2+/钙调素依赖性蛋白激酶II （CaMKII）抑制剂，以及细胞外信号调节激酶（ERK）激活剂。几种适当的化学物质组合协同增强了小鼠和人类PSCs衍生的心肌细胞的增殖，特别是小鼠心肌细胞的增殖高达14倍。我们还研究了鉴定的化学物质对不同发育阶段和物种的心肌细胞的影响。ERK激活剂和CaMKII抑制剂仅对早期发育阶段的心肌细胞有增殖作用，而GSK3和p38 MAPK抑制剂不仅在新生儿和成人心肌细胞中显著协同诱导细胞周期的再进入和进展。我们的方法成功地揭示了在不同发育阶段控制心肌细胞增殖的新分子靶点和机制，并为psc衍生的心肌细胞作为探索基于化学的心脏再生策略的有力工具。
The proliferation of cardiomyocytes is highly restricted after postnatal maturation, limiting heart regeneration. Elucidation of the regulatory machineries for the proliferation and growth arrest of cardiomyocytes is imperative. Chemical biology is efficient to dissect molecular mechanisms of various cellular events and often provide therapeutic potentials. We have been investigating cardiovascular differentiation with pluripotent stem cells (PSCs). The combination of stem cell and chemical biology can provide novel approaches to investigate the molecular mechanisms and manipulation of cardiomyocyte proliferation. To identify chemicals that regulate cardiomyocyte proliferation, we performed a screening of a defined chemical library based on proliferation of mouse PSC-derived cardiomyocytes and identified 4 chemical compound groups - inhibitors of glycogen synthase kinase-3 (GSK3), p38 mitogen-activated protein kinase (MAPK) and Ca2+/calmodulin-dependent protein kinase II (CaMKII), and activators of extracellular signal-regulated kinase (ERK). Several appropriate combinations of chemicals synergistically enhanced proliferation of cardiomyocytes derived from both mouse and human PSCs, notably up to a 14-fold increase in mouse cardiomyocytes. We also examined the effects of identified chemicals on cardiomyocytes in various developmental stages and species. Whereas ERK activators and CaMKII inhibitors showed proliferative effects only on cardiomyocytes in early developmental stages, GSK3 and p38 MAPK inhibitors substantially and synergistically induced reentry and progression of cell cycle in not only neonatal but also adult cardiomyocytes. Our approach successfully uncovered novel molecular targets and mechanisms controlling cardiomyocyte proliferation in distinct developmental stages and offered PSC-derived cardiomyocytes as a potent tool to explore chemical-based cardiac regenerative strategies.