NOTCH2 variant D1853H is mutated in two non-syndromic premature ovarian insufficiency patients from a Chinese pedigree
NOTCH2 variant D1853H is mutated in two non-syndromic premature ovarian insufficiency patients from a Chinese pedigree
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NOTCH2 变异体 D1853H 在两名来自中国血统的非综合征性卵巢早衰患者中发生突变。
DOI：
10.1186/s13048-020-00645-4
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发表时间：
2020-04-20
影响因子：
4
通讯作者：
Yin, Chenghong
中科院分区：
文献类型：
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作者：
Li, Lin;Feng, Fan;Yin, Chenghong
卵巢功能不全（POI）是一种严重的女性不孕症，以40岁前4-6个月闭经为特征，伴有促卵泡激素（FSH）水平升高（约25 IU/L）。虽然已经报道了几个基因参与POI的遗传基础，但POI的分子机制尚不清楚。方法全外显子组测序（WES）。进行了桑格测序以验证先证者及其母亲的变异。利用计算机算法分析该变异的突变效应。蛋白质三维结构建模用于预测突变的蛋白质结构。构建载体，转染质粒，对各组进行rna测序（RNA-seq），解剖野生型（WT）和D1853H NOTCH2突变体表达组的差异表达基因。基因本体分析还用于分析差异表达基因之间富集的生物过程或途径。结果我们报告了两例来自中国血统的非综合征型POI患者。先证者（女儿）22岁时FSH水平为46 IU/L。她的月经初潮是在12岁，但她在20岁时闭经。通过WES，发现一种罕见的杂合变异(C . 5557g > C；D1853H)在NOTCH2基因中被鉴定。基因分析表明p.D1853H为致病等位基因。蛋白质三维结构建模表明，D1853H可能增强或减弱静电表面电位。通过分子分析，我们发现表达D1853H NOTCH2突变体的细胞在激活NOTCH信号通路下游靶基因方面具有相似的作用。然而，106个蛋白编码基因在表达D1853H的细胞和表达WT NOTCH2的细胞之间存在差异表达，这些基因在胶原降解、NCAM1相互作用和hdac脱乙酰组蛋白中富集，揭示了导致POI的病理机制未知。结论NOTCH2中罕见的杂合变异可能与POI有关。这一发现为研究人员和临床医生更好地了解POI的病因、分子机制和遗传咨询提供了帮助。
Background Premature ovarian insufficiency (POI) is a severe disorder of female infertility, characterized by 4-6 months of amenorrhea before the age of 40 years, with elevated follicle stimulating hormone (FSH) levels (> 25 IU/L). Although several genes have been reported to contribute to the genetic basis of POI, the molecular mechanism of POI remains unclear. Methods Whole-exome sequencing (WES) was performed. Sanger sequencing was carried out to validate the variant in the proband and her mother. In silico algorithms were used to analyze the mutational effect of the variant. Protein 3D structural modeling was used for predicting mutated protein structures. Vector construction and plasmids transfection were performed, and subsequently RNA-sequencing (RNA-seq) was carried out in each group to dissect the differentially expressed genes in wild-type (WT) and D1853H NOTCH2 mutant expressing groups. Gene Ontology analysis was also used to analyze the enriched biological processes or pathways among the differentially expressed genes. Results We report two non-syndromic POI patients from a Chinese pedigree. The FSH level of the proband (the daughter) was 46 IU/L at the age of 22. Her menarche was at the age of 12, but she was amenorrhea at the age of 20. By WES, a rare heterozygous variant (c.5557G > C;p.D1853H) in the NOTCH2 gene was identified. In silico analysis suggested that p.D1853H was a pathogenic allele. Protein 3D structural modeling suggested that D1853H may enhance or weaken the electrostatic surface potential. By molecular analysis, we found that cells expressing the D1853H NOTCH2 mutant had similar effect in activating the NOTCH signaling pathway downstream target genes. However, 106 protein-coding genes were differentially expressed between D1853H expressing cells and WT NOTCH2 expressing cells, and these genes were enriched for collagen degradation, NCAM1 interactions and HDACs deacetylate histones, revealing a unknown underlying mechanism of the pathology that leads to POI. Conclusions We conclude that the rare heterozygous variant in NOTCH2 may be associated with POI. This finding provides researchers and clinicians with a better understanding of the etiology, molecular mechanism and genetic consulting of POI.