Two rare variants reveal the significance of Grainyhead-like 3 Arginine 391 underlying non-syndromic cleft palate only
Two rare variants reveal the significance of Grainyhead-like 3 Arginine 391 underlying non-syndromic cleft palate only
复制标题
两个罕见的变异揭示了 Grainyhead-like 3 精氨酸 391 仅在非综合征性腭裂中的重要性。
DOI：
10.1111/odi.14164
复制
发表时间：
2022-04-19
期刊：
影响因子：
3.8
通讯作者：
Chen，Feng
中科院分区：
文献类型：
--
作者：
Huang，Wenbin;He，Qing;Chen，Feng
目的非综合征性腭裂（NSCPO）是最常见的颅面先天性缺陷之一，其遗传病因尚未确定。已经证实Grainyhead‐like 3 （GRHL3）在NSCPO的发病机制中起重要作用。本研究旨在确定和验证中国队列中首次报道的导致NSCPO的grhl3变异。方法对1例中国非scpo患者进行全外显子组测序（WES），鉴定出一种罕见的grhl3变异（p.a g391 his）。引入经验证的有害变异p.a arg391cys作为阳性对照。通过斑马鱼胚胎注射、报告基因测定、活细胞成像和RNA测序来检测变异的致病性。结果斑马鱼胚胎显微注射实验表明，过表达这些变异会破坏斑马鱼胚胎的正常发育。报道者实验显示arg391干扰GRHL3的转录活性，并发挥显性负作用。有趣的是，Arg391His和Arg391Cys在活细胞成像中显示出与野生型GRHL3不同的核定位模式。大量RNA测序表明，这两个变体改变了基因表达模式。总之，本研究鉴定并表征了一种罕见的GRHL3变异，揭示了精氨酸391在GRHL3中的关键作用。我们的研究结果将有助于促进对非scpo的理解和遗传咨询。
ObjectivesNon‐syndromic cleft palate only (NSCPO) is one of the most common craniofacial birth defects with largely undetermined genetic etiology. It has been established that Grainyhead‐like 3 (GRHL3) plays an essential role in the pathogenesis of NSCPO. This study aimed to identify and verify the first‐reportedGRHL3variant underlying NSCPO among the Chinese cohort.MethodsWe performed whole‐exome sequencing (WES) on a Chinese NSCPO patient and identified a rare variant ofGRHL3(p.Arg391His). A validated deleterious variant p.Arg391Cys was introduced as a positive control. Zebrafish embryos injection, reporter assays, live‐cell imaging, and RNA sequencing were conducted to test the pathogenicity of the variants.ResultsZebrafish embryos microinjection demonstrated that overexpression of the variants could disrupt the normal development of zebrafish embryos. Reporter assays showed that Arg391His disturbed transcriptional activity of GRHL3 and exerted a dominant‐negative effect. Interestingly, Arg391His and Arg391Cys displayed distinct nuclear localization patterns from that of wild‐type GRHL3 in live‐cell imaging. Bulk RNA sequencing suggested that the two variants changed the pattern of gene expression.ConclusionsIn aggregate, this study identified and characterized a rareGRHL3variant in NSCPO, revealing the critical role of Arginine 391 in GRHL3. Our findings will help facilitate understanding and genetic counseling of NSCPO.