Song Yangliu, Chen Weicheng, Huang Zitong, Tian Guixiang, Li Mengru, Zhao Zhengshan, Feng Zhiyu, Wu Feizhen, Qian Maoxiang, Ma Xiaojing, Sheng Wei, Huang Guoying
Institutes of Biomedical Sciences, and Children's Hospital of Fudan University, Shanghai, China.
Shanghai Key Laboratory of Birth Defects, Shanghai, China.
Front Cell Dev Biol. 2021 Jan 28;9:631942. doi: 10.3389/fcell.2021.631942. eCollection 2021.
As a key component in the NOTCH signaling pathway, HES1 plays an important role in vertebrate heart development. Variants in the HES1 coding sequence are known to be associated with congenital heart disease (CHD). However, little is known about HES1 non-coding sequence variants and their association with the risk of developing CHD. We initially analyzed the non-coding sequence of the HES1 gene in 12 unrelated CHD families by direct sequencing and identified a previously unreported promoter region variant (NM_005524.4: c.-1279-1278 insAC, rs148941464) in the HES1 gene in four CHD families. The homozygous variant in patients was inherited from carrier parents with normal phenotypes, indicating a likely recessive genetic model. Given that the HES1 gene is predicted to be likely to exhibit haploinsufficiency (%HI: 11.44), we hypothesized that the HES1 homozygous variant is a genetic risk factor underlying CHD. We then carried out sequencing of this HES1 variant in 629 sporadic non-syndromic CHD cases and 696 healthy controls and performed association analysis. Interestingly, we observed a significant association of the homozygous HES1 promoter variant with CHD (18.92% of cases vs. 9.91% of controls; OR: 2.291, 95% CI: 1.637-3.207, = 9.72 × 10). No significant association with CHD was observed for the HES1 promoter heterozygous variant ( > 0.05). However, association analysis tests of the HES1 homozygous variant with each subtype of CHD revealed that this homozygous variant was strongly associated with transposition of the great arteries (TGA) (OR: 3.726, 95% CI: 1.745-7.956, = 0.0003). Moreover, the prevalence of HES1 homozygous variants in CHD patients with TGA (27.66%) was significantly higher than that in patients with other CHD subtypes or controls. Similar results were observed in a replication group of TGA ( = 64). Functional studies demonstrated that the homozygous variant in the HES1 promoter can disrupt its ability to bind RXRA, an inhibitory transcription factor, which results in abnormally high expression of the HES1 gene, indicating that this variant harbors gain-of-function effects. Our findings reveal that the non-coding homozygous variant in the HES1 promoter has a gain-of-function effect and is associated with an increased risk of CHD development, especially the severe TGA subtype.
作为NOTCH信号通路的关键组成部分，HES1在脊椎动物心脏发育中起着重要作用。已知HES1编码序列中的变异与先天性心脏病（CHD）相关。然而，关于HES1非编码序列变异及其与患CHD风险的关联却知之甚少。我们最初通过直接测序分析了12个无亲缘关系的CHD家族中HES1基因的非编码序列，并在4个CHD家族中鉴定出HES1基因中一个先前未报道的启动子区域变异（NM_005524.4:c.-1279-1278 insAC，rs148941464）。患者中的纯合变异是从表型正常的携带者父母遗传而来，表明可能是隐性遗传模式。鉴于预测HES1基因可能表现出单倍体不足（%HI:11.44），我们假设HES1纯合变异是CHD潜在的遗传危险因素。然后我们对629例散发性非综合征性CHD病例和696例健康对照进行了该HES1变异的测序，并进行了关联分析。有趣的是，我们观察到HES1启动子纯合变异与CHD存在显著关联（病例组为18.92%，对照组为9.91%；OR:2.291，95%CI:1.637 - 3.207，=9.72×10）。未观察到HES1启动子杂合变异与CHD有显著关联（>0.05）。然而，HES1纯合变异与CHD各亚型的关联分析测试表明，该纯合变异与大动脉转位（TGA）密切相关（OR:3.726，95%CI:1.745 - 7.956，=0.0003）。此外，TGA的CHD患者中HES1纯合变异的患病率（27.66%）显著高于其他CHD亚型患者或对照组。在TGA的复制组（=64）中也观察到了类似结果。功能研究表明，HES1启动子中的纯合变异会破坏其与抑制性转录因子RXRA结合的能力，从而导致HES1基因异常高表达, 表明该变异具有功能获得效应。我们的研究结果表明，HES1启动子中的非编码纯合变异具有功能获得效应，并与CHD发生风险增加相关，尤其是严重的TGA亚型。
