PDF(Pubmed)
Abstract：
BACKGROUND: Cystic fibrosis (CF) is an autosomal recessive disorder rarely found in Asian populations. Most males with CF are infertile because of obstructive azoospermia (OA) caused by congenital bilateral absence of the vas deferens (CBAVD). Compound heterozygous mutations of cystic fibrosis transmembrane conductance regulator (CFTR) are among the most common pathogenic factors in CBAVD. However, few genealogical analyses have been performed.
METHODS: In this study, whole-exome sequencing and cosegregation analysis were performed in a Chinese pedigree involving two siblings with CBAVD. Moreover, in vitro gene expressions were used to analyze the pathogenicity of a novel CFTR mutation.
RESULTS: We identified compound heterozygous mutations of CFTR comprising the known disease-causing variant c.1210-11T>G (also known as IVS9-5 T) and c.2144delA;p.q715fs in two siblings with CBAVD. To verify the effects in vitro, we transfected vectors expressing wild-type and mutated CFTR into 293T cells. The results showed that the CFTR protein containing the frameshift mutation (c.2144delA) was 60 kD smaller. With testicular sperm aspiration/intracytoplasmic sperm injection-embryo transfer (TESA/ICSI-ET), both CBAVD patients fathered healthy offspring.
CONCLUSIONS: Our study revealed that compound heterozygous mutations of CFTR are involved in CBAVD, expanding the known CFTR gene mutation spectrum of CBAVD patients and providing more evidence that compound heterozygous mutations can cause familial CBAVD.
METHODS: In this study, whole-exome sequencing and cosegregation analysis were performed in a Chinese pedigree involving two siblings with CBAVD. Moreover, in vitro gene expressions were used to analyze the pathogenicity of a novel CFTR mutation.
RESULTS: We identified compound heterozygous mutations of CFTR comprising the known disease-causing variant c.1210-11T>G (also known as IVS9-5 T) and c.2144delA;p.q715fs in two siblings with CBAVD. To verify the effects in vitro, we transfected vectors expressing wild-type and mutated CFTR into 293T cells. The results showed that the CFTR protein containing the frameshift mutation (c.2144delA) was 60 kD smaller. With testicular sperm aspiration/intracytoplasmic sperm injection-embryo transfer (TESA/ICSI-ET), both CBAVD patients fathered healthy offspring.
CONCLUSIONS: Our study revealed that compound heterozygous mutations of CFTR are involved in CBAVD, expanding the known CFTR gene mutation spectrum of CBAVD patients and providing more evidence that compound heterozygous mutations can cause familial CBAVD.
摘要：
背景：囊性纤维化（CF）是一种在亚洲人群中很少发现的常染色体隐性遗传疾病。大多数患有CF的男性由于先天性双侧输精管缺失（CBAVD）引起的阻塞性无精子症（OA）而不育。囊性纤维化跨膜传导调节因子（CFTR）的复合杂合突变是CBAVD中最常见的致病因素之一。然而,很少进行系谱分析。
方法：在本研究中，在一个涉及两个CBAVD兄弟姐妹的中国家系中进行了全外显子组测序和共分离分析.此外，体外基因表达用于分析新的CFTR突变的致病性。
结果：我们鉴定了CFTR的复合杂合突变，包括已知的致病变体c.1210-11T>G（也称为IVS9-5T）和c.2144delA；p。q715fs在两个兄弟姐妹与CBAVD。为了验证其体外效果，我们将表达野生型和突变CFTR的载体转染到293T细胞中。结果表明，含有移码突变（c.2144delA）的CFTR蛋白小60kD。睾丸精子抽吸/胞浆内精子注射-胚胎移植（TESA/ICSI-ET），两名CBAVD患者都是健康后代的父亲。
结论：我们的研究表明CFTR的复合杂合突变与CBAVD有关，扩大了CBAVD患者已知的CFTR基因突变谱,为复合杂合突变可引起家族性CBAVD提供了更多证据。
方法：在本研究中，在一个涉及两个CBAVD兄弟姐妹的中国家系中进行了全外显子组测序和共分离分析.此外，体外基因表达用于分析新的CFTR突变的致病性。
结果：我们鉴定了CFTR的复合杂合突变，包括已知的致病变体c.1210-11T>G（也称为IVS9-5T）和c.2144delA；p。q715fs在两个兄弟姐妹与CBAVD。为了验证其体外效果，我们将表达野生型和突变CFTR的载体转染到293T细胞中。结果表明，含有移码突变（c.2144delA）的CFTR蛋白小60kD。睾丸精子抽吸/胞浆内精子注射-胚胎移植（TESA/ICSI-ET），两名CBAVD患者都是健康后代的父亲。
结论：我们的研究表明CFTR的复合杂合突变与CBAVD有关，扩大了CBAVD患者已知的CFTR基因突变谱,为复合杂合突变可引起家族性CBAVD提供了更多证据。
