Signal Transduction and Targeted Therapy ( IF 40.8 ) Pub Date : 2024-09-06 , DOI: 10.1038/s41392-024-01942-w
Zheyi Lin 1, 2, 3, 4, 5, 6 , Bowen Lin 1, 2, 3, 4, 5, 6 , Chengwen Hang 1, 2, 3, 4, 5, 6 , Renhong Lu 1, 2, 3, 4, 5, 6 , Hui Xiong 1, 2, 3, 4, 6, 7 , Junyang Liu 1, 2, 3, 4, 6, 7 , Siyu Wang 1, 2 , Zheng Gong 1, 2 , Mingshuai Zhang 1, 2, 3, 4, 6, 7 , Desheng Li 1, 2, 3, 4, 6 , Guojian Fang 1, 2, 3, 4, 5, 6 , Jie Ding 1, 2, 3, 4, 5, 6 , Xuling Su 1, 2 , Huixin Guo 8 , Dan Shi 1, 2, 3, 4, 6 , Duanyang Xie 1, 2, 3, 4, 5, 6 , Yi Liu 1, 2, 3, 4, 5, 6 , Dandan Liang 1, 2, 3, 4, 5, 6, 9 , Jian Yang 1, 2, 3, 4, 6, 7, 9 , Yi-Han Chen 1, 2, 3, 4, 5, 6, 9
Cardiac biological pacing (BP) is one of the future directions for bradyarrhythmias intervention. Currently, cardiac pacemaker cells (PCs) used for cardiac BP are mainly derived from pluripotent stem cells (PSCs). However, the production of high-quality cardiac PCs from PSCs remains a challenge. Here, we developed a cardiac PC differentiation strategy by adopting dual PC markers and simulating the developmental route of PCs. First, two PC markers, Shox2 and Hcn4, were selected to establish Shox2:EGFP; Hcn4:mCherry mouse PSC reporter line. Then, by stepwise guiding naïve PSCs to cardiac PCs following naïve to formative pluripotency transition and manipulating signaling pathways during cardiac PCs differentiation, we designed the FSK method that increased the yield of SHOX2+; HCN4+ cells with typical PC characteristics, which was 12 and 42 folds higher than that of the embryoid body (EB) and the monolayer M10 methods respectively. In addition, the in vitro cardiac PCs differentiation trajectory was mapped by single-cell RNA sequencing (scRNA-seq), which resembled in vivo PCs development, and ZFP503 was verified as a key regulator of cardiac PCs differentiation. These PSC-derived cardiac PCs have the potential to drive advances in cardiac BP technology, help with the understanding of PCs (patho)physiology, and benefit drug discovery for PC-related diseases as well.
中文翻译:
从小鼠多能干细胞产生高质量心脏起搏细胞的新范例
心脏生物起搏(BP)是缓慢性心律失常干预的未来方向之一。目前,用于心脏血压的心脏起搏细胞(PC)主要来源于多能干细胞(PSC)。然而,利用 PSC 生产高质量的心脏 PC 仍然是一个挑战。在这里,我们通过采用双PC标记并模拟PC的发育路线,制定了心脏PC分化策略。首先,选择两个PC标记Shox2和Hcn4 ,建立Shox2:EGFP; Hcn4:mCherry小鼠 PSC 报告基因系。然后,通过在幼稚PSC向形成性多能性转变后逐步引导幼稚PSC向心脏PC,并在心脏PC分化过程中操纵信号通路,我们设计了提高SHOX2 +产量的FSK方法; HCN4 +细胞具有典型的PC特征,分别比胚状体(EB)和单层M10方法高12倍和42倍。此外,通过单细胞RNA测序(scRNA-seq)绘制了体外心脏PC的分化轨迹,类似于体内PC的发育,并且ZFP503被验证为心脏PC分化的关键调节因子。这些源自 PSC 的心脏 PC 有潜力推动心脏血压技术的进步,有助于了解 PC(病理)生理学,并有利于 PC 相关疾病的药物发现。