非梗阻性无精子症新致病基因FER1L6在精子发生障碍中的作用及机制研究

非梗阻性无精子症（NOA）是男性不育症中最严重的类型，然而大多患者病因不明，遗传因素是重要病因之一。前期我们通过全外显子测序，在2例NOA患者中分别发现了睾丸内高表达基因FER1L6的纯合和复合杂合突变。此前有报道FER1L6同源蛋白Ferlin在线虫及果蝇精子发生和卵子发生中发挥重要作用，而且FER1L6突变可能与人类卵巢早衰有关，但其在NOA中的作用尚未见报道。基于前期发现，本课题拟开展以下工作：（1）构建FER1L6基因敲除小鼠模型，进一步证实FER1L6在精子发生中的作用；（2）利用FER1L6基因突变患者和敲除小鼠模型，综合运用生物信息学分析以及各种细胞生物学技术，探索FER1L6参与精子发生的分子调控网络；（3）调查大样本的NOA患者及对照组FER1L6基因突变情况。以期从多层次、多水平阐明FER1L6在精子发生的作用及机制，为NOA的基因诊断与临床治疗提供新的分子靶点。

Non-obstructive azoospermia (NOA) is the most severe subtype of male infertility. Although the etiology remains elusive, the genetic factor is considered to be one of the most critical driving forces of NOA. In previous studies, we found homozygous and compound heterozygous deleterious variants of FER1L6 gene, which is highly expressed in testis, in two NOA patients by whole exome sequencing. It is reported that FER1L6 homologous protein Ferlin plays an important role in spermatogenesis and oogenesis in C. elegans and Drosophila, and FER1L6 mutations are involved in premature ovarian failure in human, while its functions in male NOA are still unclear. Based on preliminary results, we propose to: ① construct FER1L6 knockout mice and validate the crucial role of FER1L6 in spermatogenesis; ② explore the molecular regulatory network of spermatogenesis including FER1L6 by using FER1L6 knockout mice and patients with FER1L6 mutations, performing bioinformatics analysis and a series of cell biology assays; ③ investigate the frequency of FER1L6 gene mutations via a large sample comparison between idiopathic NOA and normal population. Our study will elucidate the role of FER1L6 in spermatogenetic, and provide a potential target for NOA gene diagnosis and clinical treatment.