CFAP65基因缺陷导致弱畸精子症和ICSI失败的分子机制

弱畸精子症是导致男性不育的重要因素，先前研究发现这一类疾病与遗传因素有关。申请人前期研究发现CFAP65基因突变导致严重的弱畸精子症（包括精子鞭毛和顶体结构异常）及ICSI结局失败。初步的蛋白组学分析也证实精子发生过程中顶体相关蛋白表达减弱。但是，CFAP65基因变异导致弱畸精子症和ICSI失败的分子机制尚不清楚。本研究拟：1）采用Cfap65基因敲除小鼠，通过小鼠模型上基于大量胚胎统计的ICSI分析，结合合子基因组激活、DNA损伤检测等实验，探究Cfap65相关的精子畸形对早期胚胎发育的影响。2）通过精子细胞及胚胎的转录组、蛋白质组学分析，明确Cfap65基因突变导致弱畸精子症、胚胎发育不良的关键分子机制。3）利用人工卵母细胞激活、mRNA显微注射等技术尝试挽救小鼠中Cfap65基因缺陷导致的不良ICSI结局。本研究项目可为ICSI结局不良的弱畸精子症的研究和干预措施建立提供新的思路。

Asthenoteratozoospermia is one of the main factors causing male infertility and has been found to be a disorder of genetic origin according to previous studies. Our recent study showed that deleterious mutations in CFAP65 can induce severe asthenoteratozoospermia (including flagellum malformations and acrosome hypoplasia) and failed ICSI outcome. Preliminary proteomic analysis also confirmed the decreased expression of acrosomal associated proteins during spermatogenesis. However, the molecular mechanisms underlying CFAP65 associated asthenoteratozoospermia and failed ICSI outcome are still unclear. This study is designed as follows: 1) Cfap65 mutated male mice will be used to investigate the effect of Cfap65 related sperm malformations on early embryo development through ICSI analysis based on a large number of embryo statistics, combined with zygotic genome activation and DNA damage detections. 2) Transcriptome and proteomic analyses of sperm cells and embryos will also be combined to reveal the key molecular mechanisms of abnormal spermatogenesis and failed ICSI outcome caused by Cfap65 mutations. 3) Artificial oocyte activation and mRNA microinjection will be adopted to try to rescue the failed ICSI outcome caused by the defect in Cfap65. This research project will provide new ideas for the study of asthenoteratozoospermia with poor ICSI outcomes and the establishment of interventions.