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Analysis for Molecular Mechanism of Cardiomyocyte Differentiation by Using a Cell Line Which Efficiently Differentiates into Cardiomyocytes

利用高效分化为心肌细胞的细胞系分析心肌细胞分化的分子机制

基本信息

批准号：
12835003
负责人：
HAYASHI Doubun
金额：
$ 2.3万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12835003/
关键词：
Regeneration Medicine BMP Smad TAK1 ATF-2 Transcription Factor Cardiomyocyte Differentiation P19CL6 TAk1 協調作用

项目摘要

The aims of basic research for understanding the molecular mechanism of cardiomyocyte differentiation and cardiac development include the establishment of the genetic technique by which cardiomyocyte differentiation can be induced, which may contribute to the practical therapeutic application for tissue regeneration and the subsequent gene-therapy for heart failure. From this respect, we investigated the molecular mechanism by which cardiomyocyte differentiation is regulated by using a cardiomyogenic cell line, P19CL6, which can differentiate efficiently into beating cardiomyocytes.We previously demonstrated that bone morphogenetic proteins (BMPs) induce cardiornyocyte differentiation through the MAPKKK TAK1. Transcription factors Smads have been reported to play a critical role in BMP signaling and the ATF/CREB family transcription factor ATF-2 has recently been shown to be a common nuclear target of the Smad and the TAK1 pathways. We here examined whether Smads and ATF-2 are involved … More in cardiomyocyte differentiation using P19CL6, a clonal derivative of murine P19 cells. Although P19CL6 efficiently differentiates into cardiomyocytes when treated with dimethyl sulfbxide, P19CL6noggin, a P19CL6 cell line constitutively overexpressing the BMP antagonist noggin, did not differentiate into cardiomyocytes. Co-overexpression of Smadl, a ligand-specific Smad, and Smad4, a common mediator Smad, restored the ability of P19CL6noggin to differentiate into cardiomyocytes, whereas stable overexpression of Smad6, an inhibitory Smad, completely blocked differentiation of P19CL6, suggesting that the Smad pathway is necessary for cardiomyocyte differentiation. ATF-2 stimulated βMHC promoter acitvity by the synergistic manner with Smad1/4 and TAK1, while overexpression of the dominant negative form of ATF-2 reduced the promoter activities of several cardiac-specific genes and inhibited cardiomyocyte differentiation of P19CL6. These results suggest that Smads, TAK1, and their common target ATF-2 cooperatively play a critical role in cardiomyocyte differentiation. Less

基础研究的目的是了解心肌细胞分化和心脏发育的分子机制，包括建立诱导心肌细胞分化的遗传技术，这可能有助于组织再生的实际治疗应用和随后的心力衰竭基因治疗。从这个角度出发，我们研究了心肌细胞分化的分子机制，通过使用心肌细胞系，P19 CL 6，它可以有效地分化为跳动的cardiomyocytes.我们以前证明，骨形态发生蛋白（BMP）诱导心肌细胞分化通过MAPKKK TAK 1。据报道，转录因子Smads在BMP信号传导中起关键作用，ATF/CREB家族转录因子ATF-2最近已被证明是Smad和TAK 1通路的共同核靶点。我们在这里检查是否Smads和ATF-2参与 ...更多信息在心肌细胞分化中使用P19 CL 6，鼠P19细胞的克隆衍生物。虽然P19 CL 6有效地分化成心肌细胞时，用二甲基亚砜，P19 CL 6头蛋白，组成型过表达的BMP拮抗剂头蛋白的P19 CL 6细胞系，没有分化成心肌细胞。Smad 1（配体特异性的Smad）和Smad 4（共同介质Smad）的共过表达恢复了P19 CL 6 noggin向心肌细胞分化的能力，而Smad 6（抑制性的Smad）的稳定过表达完全阻断了P19 CL 6的分化，这表明Smad通路是心肌细胞分化所必需的。ATF-2通过与Smad 1/4和TAK 1的协同作用激活βMHC启动子活性，而过表达显性负性ATF-2则降低了多个心肌特异性基因的启动子活性，抑制了P19 CL 6的心肌分化。这些结果表明，Smads，TAK 1，和他们的共同目标ATF-2合作在心肌细胞分化中发挥关键作用。少