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Abstract：
BACKGROUND: ELMSAN1 (ELM2-SANT domain-containing scaffolding protein 1) is a newly identified scaffolding protein of the MiDAC (mitotic deacetylase complex), playing a pivotal role in early embryonic development. Studies on Elmsan1 knockout mice showed that its absence results in embryo lethality and heart malformation. However, the precise function of ELMSAN1 in heart development and formation remains elusive. To study its potential role in cardiac lineage, we employed human-induced pluripotent stem cells (hiPSCs) to model early cardiogenesis and investigated the function of ELMSAN1.
RESULTS: We generated ELMSAN1-deficient hiPSCs through knockdown and knockout techniques. During cardiac differentiation, ELMSAN1 depletion inhibited pluripotency deactivation, decreased the expression of cardiac-specific markers, and reduced differentiation efficiency. The impaired expression of genes associated with contractile sarcomere structure, calcium handling, and ion channels was also noted in ELMSAN1-deficient cardiomyocytes derived from hiPSCs. Additionally, through a series of structural and functional assessments, we found that ELMSAN1-null hiPSC cardiomyocytes are immature, exhibiting incomplete sarcomere Z-line structure, decreased calcium handling, and impaired electrophysiological properties. Of note, we found that the cardiac-specific role of ELMSAN1 is likely associated with histone H3K27 acetylation level. The transcriptome analysis provided additional insights, indicating maturation reduction with the energy metabolism switch and restored cell proliferation in ELMSAN1 knockout cardiomyocytes.
CONCLUSIONS: In this study, we address the significance of the direct involvement of ELMSAN1 in the differentiation and maturation of hiPSC cardiomyocytes. We first report the impact of ELMSAN1 on multiple aspects of hiPSC cardiomyocyte generation, including cardiac differentiation, sarcomere formation, calcium handling, electrophysiological maturation, and proliferation.
RESULTS: We generated ELMSAN1-deficient hiPSCs through knockdown and knockout techniques. During cardiac differentiation, ELMSAN1 depletion inhibited pluripotency deactivation, decreased the expression of cardiac-specific markers, and reduced differentiation efficiency. The impaired expression of genes associated with contractile sarcomere structure, calcium handling, and ion channels was also noted in ELMSAN1-deficient cardiomyocytes derived from hiPSCs. Additionally, through a series of structural and functional assessments, we found that ELMSAN1-null hiPSC cardiomyocytes are immature, exhibiting incomplete sarcomere Z-line structure, decreased calcium handling, and impaired electrophysiological properties. Of note, we found that the cardiac-specific role of ELMSAN1 is likely associated with histone H3K27 acetylation level. The transcriptome analysis provided additional insights, indicating maturation reduction with the energy metabolism switch and restored cell proliferation in ELMSAN1 knockout cardiomyocytes.
CONCLUSIONS: In this study, we address the significance of the direct involvement of ELMSAN1 in the differentiation and maturation of hiPSC cardiomyocytes. We first report the impact of ELMSAN1 on multiple aspects of hiPSC cardiomyocyte generation, including cardiac differentiation, sarcomere formation, calcium handling, electrophysiological maturation, and proliferation.
摘要：
背景：ELMSAN1（含ELM2-SANT结构域的支架蛋白1）是MiDAC（有丝分裂去乙酰化酶复合物）的一种新鉴定的支架蛋白，在早期胚胎发育中起着举足轻重的作用。对Elmsan1基因敲除小鼠的研究表明，它的缺失会导致胚胎死亡和心脏畸形。然而,ELMSAN1在心脏发育和形成中的精确功能仍然难以捉摸。为了研究它在心脏谱系中的潜在作用，我们采用人诱导的多能干细胞(hiPSCs)建立早期心脏发生模型,并研究了ELMSAN1的功能.
结果：我们通过敲低和敲除技术产生了ELMSAN1缺陷型hiPSCs。在心脏分化过程中,ELMSAN1耗竭抑制了多能性失活，降低了心脏特异性标志物的表达，并降低了差异化效率。与收缩肌节结构相关的基因表达受损，钙处理，在源自hiPSC的ELMSAN1缺陷型心肌细胞中也注意到离子通道。此外,通过一系列结构和功能评估，我们发现ELMSAN1-nullhiPSC心肌细胞是不成熟的，表现出不完整的肌节Z线结构，减少钙处理，和受损的电生理特性。值得注意的是，我们发现ELMSAN1的心脏特异性作用可能与组蛋白H3K27乙酰化水平相关.转录组分析提供了更多的见解，表明在ELMSAN1敲除的心肌细胞中，随着能量代谢开关的成熟减少和恢复的细胞增殖。
结论：在这项研究中，我们讨论了ELMSAN1直接参与hiPSC心肌细胞分化和成熟的重要性。我们首先报道了ELMSAN1对hiPSC心肌细胞生成的多个方面的影响，包括心脏分化，肌节形成,钙处理，电生理成熟，和扩散。
结果：我们通过敲低和敲除技术产生了ELMSAN1缺陷型hiPSCs。在心脏分化过程中,ELMSAN1耗竭抑制了多能性失活，降低了心脏特异性标志物的表达，并降低了差异化效率。与收缩肌节结构相关的基因表达受损，钙处理，在源自hiPSC的ELMSAN1缺陷型心肌细胞中也注意到离子通道。此外,通过一系列结构和功能评估，我们发现ELMSAN1-nullhiPSC心肌细胞是不成熟的，表现出不完整的肌节Z线结构，减少钙处理，和受损的电生理特性。值得注意的是，我们发现ELMSAN1的心脏特异性作用可能与组蛋白H3K27乙酰化水平相关.转录组分析提供了更多的见解，表明在ELMSAN1敲除的心肌细胞中，随着能量代谢开关的成熟减少和恢复的细胞增殖。
结论：在这项研究中，我们讨论了ELMSAN1直接参与hiPSC心肌细胞分化和成熟的重要性。我们首先报道了ELMSAN1对hiPSC心肌细胞生成的多个方面的影响，包括心脏分化，肌节形成,钙处理，电生理成熟，和扩散。
