Yunnan Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan, PR China; Department of Cardiovascular Surgery, the First Peoples' Hospital of Yunnan Province, Kunming, Yunnan, PR China.
Department of Cardiology, The First Peoples' Hospital of Yunnan Province, Kunming, Yunnan, PR China.
Mech Dev. 2020 Jun;162:103611. doi: 10.1016/j.mod.2020.103611. Epub 2020 May 3.
Mutations affecting cardiac structural genes can lead to congenital heart diseases (CHDs). Axonemal Central Pair Apparatus Protein (HYDIN) is a ciliary protein previously linked to congenital cardiomyopathy. However, the role of HYDIN in the aetiology of CHDs is thus far unknown. Herein, we explore the function of HYDIN in heart development and CHDs.
The function of HYDIN in cardiac differentiation was assessed in vitro using HYDIN siRNAs, HYDIN overexpression, and HYDIN short hairpin RNA (shRNA)-GATA binding protein 4 (GATA4) cDNA rescue constructs in the human embryonic stem cell (hESC) line HES3. To assess Hydin's function in vivo, we generated shRNA-mediated Hydin knockdown transgenic mice. We characterized the functional mechanisms of the most common human HYDIN variant associated with atrial septal defect (ASD) risk (71098693 mutant, c.A2207C) in cardiac-differentiating HES3 cells.
HYDIN functions as a positive regulator of human cardiomyocyte differentiation and promotes expression of cardiac contractile genes in hESC cells. This is mediated through GATA4, a critical transcription factor in heart development. Cardiac-specific Hydin knockdown in vivo leads to Gata4 downregulation and enhanced atrial septal defect (ASD) risk in mice. The c.A2207C HYDIN mutation reduces GATA4 expression in hESC cells.
HYDIN loss-of-function inhibits GATA4 expression and enhances ASD risk. We also establish the regulation of a key transcription factor in heart development by a ciliary protein.
影响心脏结构基因的突变可导致先天性心脏病（CHD）。轴丝中央对附属蛋白（HYDIN）是一种以前与先天性心肌病相关的纤毛蛋白。然而，HYDIN 在 CHD 发病机制中的作用迄今尚不清楚。在此，我们探讨了 HYDIN 在心脏发育和 CHD 中的作用。
在人类胚胎干细胞（hESC）系 HES3 中，使用 HYDIN siRNA、HYDIN 过表达和 HYDIN 短发夹 RNA（shRNA）-GATA 结合蛋白 4（GATA4）cDNA 挽救构建体，体外评估 HYDIN 在心脏分化中的功能。为了评估 Hydin 在体内的功能，我们生成了 shRNA 介导的 Hydin 敲低转基因小鼠。我们在心脏分化的 HES3 细胞中对与房间隔缺损（ASD）风险相关的最常见人类 HYDIN 变体（71098693 突变，c.A2207C）的功能机制进行了特征描述。
HYDIN 作为人类心肌细胞分化的正调节剂，可促进 hESC 细胞中心脏收缩基因的表达。这是通过 GATA4 介导的，GATA4 是心脏发育的关键转录因子。体内心脏特异性 Hydin 敲低导致 Gata4 下调，并增加小鼠的 ASD 风险。HYDIN 的 c.A2207C 突变降低了 hESC 细胞中的 GATA4 表达。
HYDIN 功能丧失抑制 GATA4 表达并增强 ASD 风险。我们还建立了纤毛蛋白对心脏发育关键转录因子的调节。
