Biallelic mutations in CFAP65 cause male infertility with multiple morphological abnormalities of the sperm flagella in humans and mice
Biallelic mutations in CFAP65 cause male infertility with multiple morphological abnormalities of the sperm flagella in humans and mice
复制标题
CFAP65的双等位基因突变导致男性不育，并导致人类和小鼠精子鞭毛出现多种形态异常
DOI：
10.1136/jmedgenet-2019-106344
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发表时间：
2020-02-01
影响因子：
4
通讯作者：
Zhang, Feng
中科院分区：
文献类型：
--
作者：
Li, Weiyu;Wu, Huan;Zhang, Feng
背景男性不育是一个世界性的普遍问题，主要是由于精子活力受损。精子鞭毛的多种形态异常（MMAF）表现为异常精子，鞭毛缺失、短、卷曲、弯曲和口径不规则，导致活动力严重下降。以前的研究报告了几个MMAF相关基因占约一半的MMAF cases.Methods和结果我们进行了遗传分析，使用全外显子组测序在88个中国汉族MMAF先证者。CFAP 65纯合突变在4个无关的近亲家庭中被发现，CFAP 65复合杂合突变在两个无关的MMAF病例中被发现。所有这些CFAP 65突变均为无效，包括4个移码突变（c.1775delC [p.Pro592Leufs*8]，c.3072_3079dup [p.Arg1027Profs*41]，c.1946delC [p.Pro649Argfs*5]和c.1580delT [p.Leu527Argfs*31]）和三个停止增益突变（c.4855C>T [p.Arg1619*]，c.5270T>A [p.Leu1757*]和c.5341G>T [p.Glu1781*]）。此外，在突尼斯和伊朗血统的两名MMAF受影响的男性中分别鉴定出两种可能影响剪接的纯合CFAP 65变体。CFAP 65的这些双等位基因变体通过桑格测序验证，并且在聚集来自不同人群的序列信息的大数据集中不存在或非常罕见。CFAP 65编码纤毛和鞭毛相关蛋白65，在睾丸中高度优先表达。在这里，我们还使用CRISPR-Cas9技术在小鼠直向同源物Cfap 65中产生了移码突变。值得注意的是，Cfap 65突变的雄性小鼠的表型与人类MMAF.Conclusions人类受试者和Cfap 65突变的小鼠进行的实验观察表明，CFAP 65的双等位基因突变的存在下，导致MMAF表型和损害精子活力。
Background Male infertility is a prevalent issue worldwide, mostly due to the impaired sperm motility. Multiple morphological abnormalities of the sperm flagella (MMAF) present aberrant spermatozoa with absent, short, coiled, bent and irregular-calibre flagella resulting in severely decreased motility. Previous studies reported several MMAF-associated genes accounting for approximately half of MMAF cases.Methods and result We conducted genetic analysis using whole-exome sequencing in 88 Han Chinese MMAF probands. CFAP65 homozygous mutations were identified in four unrelated consanguineous families, and CFAP65 compound heterozygous mutations were found in two unrelated cases with MMAF. All these CFAP65 mutations were null, including four frameshift mutations (c.1775delC [p.Pro592Leufs*8], c.3072_3079dup [p.Arg1027Profs*41], c.1946delC [p.Pro649Argfs*5] and c.1580delT [p.Leu527Argfs*31]) and three stop-gain mutations (c.4855C>T [p.Arg1619*], c.5270T>A [p.Leu1757*] and c.5341G>T [p.Glu1781*]). Additionally, two homozygous CFAP65 variants likely affecting splicing were identified in two MMAF-affected men of Tunisian and Iranian ancestries, respectively. These biallelic variants of CFAP65 were verified by Sanger sequencing and were absent or very rare in large data sets aggregating sequence information from various human populations. CFAP65, encoding the cilia and flagella associated protein 65, is highly and preferentially expressed in the testis. Here we also generated a frameshift mutation in mouse orthologue Cfap65 using CRISPR-Cas9 technology. Remarkably, the phenotypes of Cfap65-mutated male mice were consistent with human MMAF.Conclusions Our experimental observations performed on both human subjects and on Cfap65-mutated mice demonstrate that the presence of biallelic mutations in CFAP65 causes the MMAF phenotype and impairs sperm motility.