Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Department of Biological Statistics and Computational Biology, Cornell University, Ithaca, NY 14853, USA.
Hum Mol Genet. 2020 Dec 18;29(20):3402-3411. doi: 10.1093/hmg/ddaa230.
Approximately 7% of men worldwide suffer from infertility, with sperm abnormalities being the most common defect. Though genetic causes are thought to underlie a substantial fraction of idiopathic cases, the actual molecular bases are usually undetermined. Because the consequences of most genetic variants in populations are unknown, this complicates genetic diagnosis even after genome sequencing of patients. Some patients with ciliopathies, including primary ciliary dyskinesia and Bardet-Biedl syndrome, also suffer from infertility because cilia and sperm flagella share several characteristics. Here, we identified two deleterious alleles of RABL2A, a gene essential for normal function of cilia and flagella. Our in silico predictions and in vitro assays suggest that both alleles destabilize the protein. We constructed and analyzed mice homozygous for these two single-nucleotide polymorphisms, Rabl2L119F (rs80006029) and Rabl2V158F (rs200121688), and found that they exhibit ciliopathy-associated disorders including male infertility, early growth retardation, excessive weight gain in adulthood, heterotaxia, pre-axial polydactyly, neural tube defects and hydrocephalus. Our study provides a paradigm for triaging candidate infertility variants in the population for in vivo functional validation, using computational, in vitro and in vivo approaches.
全世界约有 7%的男性患有不育症,而精子异常是最常见的缺陷。尽管遗传原因被认为是许多特发性病例的基础,但实际的分子基础通常是不确定的。由于大多数人群中遗传变异的后果未知,这使得即使对患者进行基因组测序后,遗传诊断也变得复杂。一些纤毛病患者,包括原发性纤毛运动障碍和 Bardet-Biedl 综合征,也因纤毛和精子鞭毛具有几个共同特征而患有不育症。在这里,我们鉴定了 RABL2A 的两个有害等位基因,该基因对于纤毛和鞭毛的正常功能至关重要。我们的计算机预测和体外检测表明,这两个等位基因都使蛋白质不稳定。我们构建并分析了这两种单核苷酸多态性(Rabl2L119F [rs80006029]和 Rabl2V158F [rs200121688])的纯合子小鼠,发现它们表现出纤毛病相关疾病,包括男性不育、早期生长迟缓、成年后体重过度增加、异位症、前轴多指畸形、神经管缺陷和脑积水。我们的研究为使用计算、体外和体内方法对人群中候选不育变异进行体内功能验证提供了范例。