家族性卵巢早衰合并感音神经性耳聋（Perrault综合征）致病基因筛选及功能研究

Perrault syndrome (PS) is inherited as an autosomal recessive disease. Female patients' phenotype is ovarian dysplasia, amenorrhea, and sensorineural deafness. The pathogenesis is not clear, and research papers are limited to case reports till now. This project will focus on the causative gene and molecular mechanism of familial Perrault's syndrome（PS）. First, screening of the reported candidate genes and variants will be performed in 2 rare pedigree of familial PS; Whole-gene exome sequencing will be used if no positive results got. And then bioinformatics analysis and functional studies both in vivo and in vitro will be performed to clarify the possible pathogenic mechanism of the candidate genes and to elucidate the deleterious effect when causative mutations occurred. The results will provide new clues for identifying novel genes and related mechanism contributing to PS and ovarian function. The project would also be helpful for understanding the etiology of genetic heterogeneity and providing the important experimental and theoretical basis for clinical diagnosis of premature ovarian failure.

Perrault综合征（Perrault syndrome ，PS) 是一种临床罕见的常染色体隐性遗传病，女性患者表现为卵巢发育不良、闭经和感音神经性耳聋，其发病机制尚不清楚，目前的报道多局限于病例个案报道。本项目将重点研究家族性卵巢早衰合并感音神经性耳聋（PS）的致病基因和作用机理：首先对已收集的两个PS家系的成员进行已报道的候选基因和突变位点的筛查，若无阳性结果，则行全基因组外显子组测序初步确定候选基因和突变位点，然后通过生物信息学分析、体内外功能实验等阐明基因突变对蛋白功能及卵巢功能的影响，验证致病基因在PS发病中的作用。本项目的研究结果为发现卵巢功能相关基因、阐明PS发病机理提供新的线索，对认识卵巢早衰病因的遗传异质性和临床基因诊断提供重要的实验和理论基础。

1.我们对一例卵巢早衰（Premature ovarian failure，POF）合并感音性耳聋（Perrault综合征）家系进行全外显子测序（whole exome sequencing，WES），发现位于线粒体代谢相关基因C10ORF2纯合错意突变位点R463Q，该突变位于编码蛋白的解旋酶区域，在各物种间相对保守。功能实验证实突变R463Q能够影响线粒体的储备呼吸能力，提示该突变引发的生殖细胞呼吸能力衰减可能是POI的致病机制之一。2.首次在384例中国汉族散发POF患者中发现WT1基因错义突变p.Pro126Ser和p.Arg370His，随后通过功能实验证明此两处错义突变均可影响WT1基因对多个下游基因的转录调控，从而影响卵母细胞-颗粒细胞相互作用，导致卵巢功能衰竭。3.首次在170例中国汉族低促性腺素性功能减退症患者中对KISS1基因进行突变筛查，发现2处已知单核苷酸多态位点（Single nucleotide polymorphisms, SNPs）：rs12998 (c.58G>A)和rs4889 (c.242C>G)，SNPs位点发生率与对照人群基因型频率相比没有统计学差异，提示这两处已知SNPs位点不引起KISS1基因功能的改变，KISS1基因突变不是中国汉族促性腺素性功能减退患者的常见病因。