State Key Laboratory of Cardiovascular Diseases, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai, 200120, China.
Shanghai Arrhythmia Research Center, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai, 200120, China.
Signal Transduct Target Ther. 2024 Sep 6;9(1):230. doi: 10.1038/s41392-024-01942-w.
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.
心脏生物起搏 (BP) 是未来治疗心动过缓的方向之一。目前，用于心脏 BP 的心脏起搏器细胞 (PCs) 主要来源于多能干细胞 (PSCs)。然而，从 PSCs 中产生高质量的心脏 PCs 仍然是一个挑战。在这里，我们采用双 PC 标志物并模拟 PC 发育途径，开发了一种心脏 PC 分化策略。首先，选择两个 PC 标志物 Shox2 和 Hcn4 来建立 Shox2:EGFP; Hcn4:mCherry 小鼠 PSC 报告系。然后，通过逐步引导原始 PSCs 向心脏 PCs 分化，遵循原始到形成性多能性过渡，并在心脏 PCs 分化过程中操纵信号通路，我们设计了 FSK 方法，该方法提高了具有典型 PC 特征的 SHOX2; HCN4 细胞的产量，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 PCs 分化轨迹，该轨迹类似于体内 PCs 发育，并且验证了 ZFP503 是心脏 PCs 分化的关键调节因子。这些 PSC 来源的心脏 PCs 有可能推动心脏 BP 技术的进步，有助于理解 PCs 的 (病理)生理学，并有益于与 PCs 相关疾病的药物发现。
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