Deep sequencing analysis of MesP1 dependent cardiovascular signalling pathways during stem cell differentiation

干细胞分化过程中MesP1依赖性心血管信号通路的深度分析测序

• 批准号: 197123046
• 负责人: Professor Dr. Robert David
• 金额: --
• 依托单位: Klinik und Poliklinik für Herzchirurgie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/197123046?language=en
• 关键词: Deep; sequencing; analysis; MesP1; dependent

The proliferative potential of stem cell derived cardiomyocytes is limited and reasonable yields to repair a human infarcted heart have yet to be achieved. It is crucial to decipher the biological processes underlying cardiovascular stem cell differentiation in order to preprogram pluripotent cells for cardiovascular cell therapy and tissue engineering. We have shown that the transcription factor MesP1 represents a master regulator sufficient to induce cardiovasculogenesis in stem cells as well as vertebrate embryos. Thereby, we defined the Dkk-1 promoter as a direct target of MesP1 leading to blockage of canonical wnt-signaling. More recently we demonstrated proof of principle for cardiovascular subtype specific programming of ES cells: MesP1 forced the appearance of early/intermediate type cardiomyocytes whereas Nkx2.5 led to preferentially differentiated ventricular cells. Relying on stably and inducible MesP1 overexpressing ES cell lines here we plan the analysis of cardiovascular stem cell differentiation using a global approach based on deep sequencing at the mRNA (Transcriptome) level. Thereby MesP1 positive will be compared to MesP1 negative lateral plate mesodermal cells. In addition novel MesP1 target promoters will be identified via ChIP-Seq. Overall we expect to obtain novel important information of yet unknown signalling pathways and factors driving these events as well as to come up with novel surface markers possibly allowing purification of cardiovascular precursor cells without genetic modification.

干细胞衍生的心肌细胞的增殖潜力是有限的，并且尚未实现修复人类梗塞心脏的合理产量。因此，研究心血管干细胞分化的生物学机制，对于为心血管细胞治疗和组织工程提供多能干细胞的预编程至关重要。我们已经表明，转录因子MesP 1代表一个主调节足以诱导干细胞以及脊椎动物胚胎心血管发生。因此，我们将Dkk-1启动子定义为MesP 1的直接靶点，导致经典wnt信号传导的阻断。最近，我们证明了ES细胞的心血管亚型特异性编程的原理证明：MesP 1迫使早期/中间型心肌细胞的出现，而Nkx2.5导致优先分化的心室细胞。依靠稳定和可诱导的MesP 1过表达ES细胞系，我们计划使用基于mRNA（转录组）水平深度测序的全球方法分析心血管干细胞分化。因此，将MesP 1阳性与MesP 1阴性侧板中胚层细胞进行比较。此外，新的MesP 1靶启动子将通过ChIP-Seq鉴定。总的来说，我们希望获得新的重要信息，但未知的信号通路和因素驱动这些事件，以及拿出新的表面标记物可能允许纯化心血管前体细胞没有遗传修饰。