The homeodomain transcription factor SHOX2 is involved in the development and function of the heart's primary pacemaker, the sinoatrial node (SAN), and has been associated with cardiac conduction-related diseases such as atrial fibrillation and sinus node dysfunction. To shed light on Shox2-dependent genetic processes involved in these diseases, we established a murine embryonic stem cell (ESC) cardiac differentiation model to investigate Shox2 pathways in SAN-like cardiomyocytes. Differential RNA-seq-based expression profiling of Shox2^+/+ and Shox2^−/− ESCs revealed 94 dysregulated transcripts in Shox2^−/− ESC-derived SAN-like cells. Of these, 15 putative Shox2 target genes were selected for further validation based on comparative expression analysis with SAN- and right atria-enriched genes. Network-based analyses, integrating data from the Mouse Organogenesis Cell Atlas and the Ingenuity pathways, as well as validation in mouse and zebrafish models confirmed a regulatory role for the novel identified Shox2 target genes including Cav1, Fkbp10, Igfbp5, Mcf2l and Nr2f2. Our results indicate that genetic networks involving SHOX2 may contribute to conduction traits through the regulation of these genes.

同源结构域转录因子SHOX2参与心脏主要起搏器窦房结（SAN）的发育和功能，并与诸如心房颤动和窦房结功能障碍等与心脏传导相关的疾病有关。为了阐明与这些疾病相关的Shox2依赖性遗传过程，我们建立了小鼠胚胎干细胞（ESC）心脏分化模型，以研究SAN样心肌细胞中的Shox2途径。Shox2 ^{+ / +}和Shox2 ^-/- ESC的差异基于RNA序列的表达谱显示，Shox2 ^-/-中有94个转录失调^。ESC衍生的类SAN细胞。其中，选择15个推定的Shox2靶基因用于基于SAN和右心房富集基因的比较表达分析进行进一步验证。基于网络的分析，整合了来自小鼠器官发生细胞图谱和Ingenuity途径的数据，以及在小鼠和斑马鱼模型中的验证，证实了新型鉴定的Shox2目标基因（包括Cav1，Fkbp10，Igfbp5，Mcf2l和Nr2f2）的调控作用。我们的结果表明，涉及SHOX2的遗传网络可能通过调节这些基因来促进传导性状。