Tan X J, Huang H, He F, Zhu L, Li H, Jiang Y S, Li H, Huang X H, Sun Z S, Li Z H
Reproductive and Genetic Center of Central Hospital of Xiangtan, Xiangtan 411100, China.
Zhonghua Xin Xue Guan Bing Za Zhi. 2016 May 24;44(5):411-5. doi: 10.3760/cma.j.issn.0253-3758.2016.05.009.
To define the potential causative gene mutation in a Chinese pedigree with progressive cardiac conduction defect (PCCD).
Sanger sequencing was performed to define potential causative gene mutation in a four-generation family with 68 members including seven PCCD patients (5 male) from 2010 to 2015.No causative gene was detected by screening known candidate genes related to PCCD including SCN5A, NKX2.5 and LMNA.High-throughput sequencing technology on exon-enriched DNA was then used to search the causative genes in 2 patients and one normal family member.
Eight new non-synonymous single nucleotide variants including AQP7 gene (exon5: c.T343C: p.Y115H), CACNA1B gene (NM_001243812: exon19: c.A2986G: p.T996A), CATSPERB gene (exon27: c.C3254G: p.P1085R), CLCA2 gene (exon11: c.G1725T: p.W575C), CLCA3P gene (ncRNA_intronic), MYLK-AS1 gene (ncRNA_intronic), TTN gene (ncRNA_UTR3), LMNA gene (LMNA: NM_170708: exon5: c.C922T: p.Q308X) were identified by comparing and filtering the results with known public databases.Then, more detailed biological analysis on these 8 genes was conducted.Traditional Sanger sequencing validated the exome sequencing results, and found that the mutation c. 1725G﹥T in gene CLCA2 segregated with the phenotype of this PCCD pedigree.The mutation c. 1725G﹥T in gene CLCA2 was thus be considered as the causative PCCD gene in this pedigree from the perspective of genetics and genomics.
The heterozygote mutation c. 1725G﹥T in gene CLCA2 might be causative gene in this PCCD pedigree.This finding adds new gene mutation variant responsible for PCCD.
确定一个患有进行性心脏传导缺陷(PCCD)的中国家系中的潜在致病基因突变。
2010年至2015年,对一个包含68名成员的四代家系进行桑格测序,以确定潜在的致病基因突变,该家系中有7名PCCD患者(5名男性)。通过筛查与PCCD相关的已知候选基因,包括SCN5A、NKX2.5和LMNA,未检测到致病基因。随后,对2名患者和1名正常家庭成员的外显子富集DNA使用高通量测序技术来寻找致病基因。
通过与已知公共数据库比较和筛选结果,鉴定出8个新的非同义单核苷酸变异,包括水通道蛋白7基因(外显子5:c.T343C:p.Y115H)、电压依赖性钙通道α1B亚基基因(NM_001243812:外显子19:c.A2986G:p.T996A)、阳离子通道精子相关B基因(外显子27:c.C3254G:p.P1085R)、氯通道蛋白2基因(外显子11:c.G1725T:p.W575C)、氯通道蛋白3假基因(非编码RNA内含子)、肌球蛋白轻链激酶反义RNA1基因(非编码RNA内含子)、肌联蛋白基因(非编码RNA 3′非翻译区)、核纤层蛋白A/C基因(LMNA:NM_170708:外显子5:c.C922T:p.Q308X)。然后,对这8个基因进行了更详细的生物学分析。传统的桑格测序验证了外显子组测序结果,发现氯通道蛋白2基因中的突变c.1725G﹥T与该PCCD家系的表型共分离。因此,从遗传学和基因组学的角度来看,氯通道蛋白2基因中的突变c.1725G﹥T被认为是该家系中导致PCCD的基因。
氯通道蛋白2基因中的杂合突变c.1725G﹥T可能是该PCCD家系的致病基因。这一发现增加了导致PCCD的新基因突变变体。