Comparative expression analysis of Shox2-deficient embryonic stem cell-derived sinoatrial node-like cells
Comparative expression analysis of Shox2-deficient embryonic stem cell-derived sinoatrial node-like cells
复制标题
DOI：
10.1016/j.scr.2017.03.018
复制
发表时间：
2017-05-01
影响因子：
1.2
通讯作者：
Rappold, Gudrun A.
中科院分区：
文献类型：
--
作者：
Hoffmann, Sandra;Schmitteckert, Stefanie;Rappold, Gudrun A.
同源结构域转录因子Shox2控制着天然心脏起搏器--窦房结的发育和功能。此外，SHOX2突变与人类心律失常有关。为了详细研究Shox2依赖的SAN细胞的发育机制，我们以Shox2为分子工具，建立了基于小鼠胚胎干细胞(ESC)的模型。Shox2(+/+)和Shox2(-/-)ESC克隆按照5种不同的方法分离和分化，以评估最有效的SAN样细胞的富集率。细胞亚型特异性标记基因的表达分析显示，基于CD166的细胞分选后最有效的富集物。用NCounter技术比较Shox2(+/+)和Shox2(-/-)ESCs的心脏表达谱。在其他基因中，我们发现NPPB是ESCs分化过程中一个新的假定的Shox2靶点。可以证实NPPB在Shox2(-/-)胚胎心脏组织中的差异表达。综上所述，我们建立了一种基于胚胎干细胞的心脏分化模型，并成功地纯化了Shox2(+/+)和Shox2(-/-)SAN样细胞。这为在生理和病理生理学条件下研究分子机制和评价新的治疗方法提供了很好的基础。(C)2017年作者。爱思唯尔出版公司(Elsevier B.V.)
The homeodomain transcription factor Shox2 controls the development and function of the native cardiac pacemaker, the sinoatrial node (SAN). Moreover, SHOX2 mutations have been associated with cardiac arrhythmias in humans. For detailed examination of Shox2-dependent developmental mechanisms in SAN cells, we established a murine embryonic stem cell (ESC)-based model using Shox2 as a molecular tool. Shox2(+/+) and Shox2(-/-) ESC clones were isolated and differentiated according to five different protocols in order to evaluate the most efficient enrichment of SAN-like cells. Expression analysis of cell subtype-specific marker genes revealed most efficient enrichment after CD166-based cell sorting. Comparative cardiac expression profiles of Shox2(+/+) and Shox2(-/-) ESCs were examined by nCounter technology. Among other genes, we identified Nppb as a novel putative Shox2 target during differentiation in ESCs. Differential expression of Nppb could be confirmed in heart tissue of Shox2(-/-)embryos. Taken together, we established an ESC-based cardiac differentiation model and successfully purified Shox2(+/+) and Shox2(-/-) SAN-like cells. This now provides an excellent basis for the investigation of molecular mechanisms under physiological and pathophysiological conditions for evaluating novel therapeutic approaches. (C) 2017 The Authors. Published by Elsevier B.V.