PDF(Pubmed)
Abstract：
OBJECTIVE: Aggregating evidence convincingly establishes the predominant genetic basis underlying congenital heart defects (CHD), though the heritable determinants contributing to CHD in the majority of cases remain elusive. In the current investigation, BMP10 was selected as a prime candidate gene for human CHD mainly due to cardiovascular developmental abnormalities in Bmp10-knockout animals. The objective of this retrospective study was to identify a new BMP10 mutation responsible for CHD and characterize the functional effect of the identified CHD-causing BMP10 mutation.
METHODS: Sequencing assay of BMP10 was fulfilled in a cohort of 276 probands with various CHD and a total of 288 non-CHD volunteers. The available family members from the proband harboring an identified BMP10 mutation were also BMP10-genotyped. The effect of the identified CHD-causative BMP10 mutation on the transactivation of TBX20 and NKX2.5 by BMP10 was quantitatively analyzed in maintained HeLa cells utilizing a dual-luciferase reporter assay system.
RESULTS: A novel heterozygous BMP10 mutation, NM_014482.3:c.247G>T;p.(Glu83*), was identified in one proband with patent ductus arteriosus (PDA), which was confirmed to co-segregate with the PDA phenotype in the mutation carrier\'s family. The nonsense mutation was not observed in 288 non-CHD volunteers. Functional analysis unveiled that Glu83*-mutant BMP10 had no transactivation on its two representative target genes TBX20 and NKX2.5, which were both reported to cause CHD.
CONCLUSIONS: These findings provide strong evidence indicating that genetically compromised BMP10 predisposes human beings to CHD, which sheds light on the new molecular mechanism that underlies CHD and allows for antenatal genetic counseling and individualized precise management of CHD.
METHODS: Sequencing assay of BMP10 was fulfilled in a cohort of 276 probands with various CHD and a total of 288 non-CHD volunteers. The available family members from the proband harboring an identified BMP10 mutation were also BMP10-genotyped. The effect of the identified CHD-causative BMP10 mutation on the transactivation of TBX20 and NKX2.5 by BMP10 was quantitatively analyzed in maintained HeLa cells utilizing a dual-luciferase reporter assay system.
RESULTS: A novel heterozygous BMP10 mutation, NM_014482.3:c.247G>T;p.(Glu83*), was identified in one proband with patent ductus arteriosus (PDA), which was confirmed to co-segregate with the PDA phenotype in the mutation carrier\'s family. The nonsense mutation was not observed in 288 non-CHD volunteers. Functional analysis unveiled that Glu83*-mutant BMP10 had no transactivation on its two representative target genes TBX20 and NKX2.5, which were both reported to cause CHD.
CONCLUSIONS: These findings provide strong evidence indicating that genetically compromised BMP10 predisposes human beings to CHD, which sheds light on the new molecular mechanism that underlies CHD and allows for antenatal genetic counseling and individualized precise management of CHD.
摘要：
目的：汇总的证据令人信服地确定了先天性心脏病（CHD）的主要遗传基础，尽管在大多数情况下导致冠心病的可遗传决定因素仍然难以捉摸。在目前的调查中,BMP10被选为人类CHD的主要候选基因，主要是由于Bmp10敲除动物的心血管发育异常。这项回顾性研究的目的是鉴定引起CHD的新BMP10突变，并表征鉴定的引起CHD的BMP10突变的功能作用。
方法：在276名患有各种CHD的先证者和总共288名非CHD志愿者的队列中完成了BMP10的测序测定。来自具有鉴定的BMP10突变的先证者的可用家族成员也被BMP10基因分型。利用双荧光素酶报告基因测定系统,在维持的HeLa细胞中定量分析鉴定的CHD致病BMP10突变对BMP10对TBX20和NKX2.5的反式激活的影响。
结果：一个新的杂合BMP10突变，NM_014482.3：c.247G>T；p。(Glu83*),在一个先证者中发现动脉导管未闭（PDA），证实与突变携带者家族中的PDA表型共分离。在288名非CHD志愿者中未观察到无义突变。功能分析揭示了Glu83*-突变体BMP10对其两个代表性靶基因TBX20和NKX2.5没有反式激活，据报道这两个靶基因都会导致CHD。
结论：这些发现提供了强有力的证据，表明基因受损的BMP10使人类容易患上冠心病，这揭示了CHD背后的新分子机制，并允许产前遗传咨询和CHD的个性化精确管理。
方法：在276名患有各种CHD的先证者和总共288名非CHD志愿者的队列中完成了BMP10的测序测定。来自具有鉴定的BMP10突变的先证者的可用家族成员也被BMP10基因分型。利用双荧光素酶报告基因测定系统,在维持的HeLa细胞中定量分析鉴定的CHD致病BMP10突变对BMP10对TBX20和NKX2.5的反式激活的影响。
结果：一个新的杂合BMP10突变，NM_014482.3：c.247G>T；p。(Glu83*),在一个先证者中发现动脉导管未闭（PDA），证实与突变携带者家族中的PDA表型共分离。在288名非CHD志愿者中未观察到无义突变。功能分析揭示了Glu83*-突变体BMP10对其两个代表性靶基因TBX20和NKX2.5没有反式激活，据报道这两个靶基因都会导致CHD。
结论：这些发现提供了强有力的证据，表明基因受损的BMP10使人类容易患上冠心病，这揭示了CHD背后的新分子机制，并允许产前遗传咨询和CHD的个性化精确管理。
