Exome analysis of a family with pleiotropic congenital heart disease.
Exome analysis of a family with pleiotropic congenital heart disease.
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DOI：
10.1161/circgenetics.111.961797
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发表时间：
2012-04-01
期刊：
影响因子：
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通讯作者：
Bowles NE
中科院分区：
文献类型：
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作者：
Arrington CB;Bleyl SB;Matsunami N;Bonnell GD;Otterud BE;Nielsen DC;Stevens J;Levy S;Leppert MF;Bowles NE
在孤立性或非综合征性先天性心脏病（CHD）患者中已经发现了许多单基因缺陷。然而，由于显著的遗传异质性，候选基因方法在大多数情况下在发现高风险等位基因方面的成功有限。使用外显子组测序鉴定一个高外显多效性CHD家族的高危基因变异采用外显子组测序法对一个CHD家系的2名成员的DNA进行分析。过滤变异体以消除常见变异体和测序伪影，然后基于变异体和基因功能的预测影响进行优先排序。使用PCR、高分辨率解链分析和DNA测序来筛选家族的其余部分，以评估变体分离。筛选后，2个个体共有2000多个罕见变异（包括单核苷酸取代和插入缺失）。其中，46个是非同义的，3个被预测为改变剪接，6个导致移码。优先化将可能涉及CHD的变异数量减少到18个。在家系中，没有一种变异与CHD完全分离。然而，除了一个受影响的个体之外，在所有受影响的个体中都发现了一种变体Myh6 Ala290Pro。这种变体之前曾在一名三尖瓣闭锁和大的第二房间隔缺损患者中发现。下一代测序很可能成为解开CHD复杂遗传学的首选方法，但通过分析家庭传播获得的信息将是至关重要的。
A number of single gene defects have been identified in patients with isolated or nonsyndromic congenital heart defects (CHD). However, due to significant genetic heterogeneity candidate gene approaches have had limited success in finding high-risk alleles in most cases. Use exome sequencing to identify high-risk gene variants in a family with highly penetrant pleiotropic CHD. DNA samples from 2 members of a family with diverse CHD were analyzed by exome sequencing. Variants were filtered to eliminate common variants and sequencing artifacts and then prioritized based upon the predicted effect of the variant and on gene function. The remainder of the family was screened using PCR, high resolution melting analysis and DNA sequencing to evaluate variant segregation. After filtering, more than 2000 rare variants (including single nucleotide substitutions and indels) were shared by the 2 individuals. Of these, 46 were non-synonymous, 3 were predicted to alter splicing, and 6 resulted in a frameshift. Prioritization reduced the number of variants potentially involved in CHD to 18. None of the variants completely segregated with CHD in the kindred. However, one variant, Myh6 Ala290Pro, was identified in all but one affected individual. This variant was previously identified in a patient with tricuspid atresia and large secundum ASD. It is likely that next generation sequencing will become the method of choice for unraveling the complex genetics of CHD, but information gained by analysis of transmission through families will be crucial.