Gross BMPR2 gene rearrangements constitute a new cause for primary pulmonary hypertension
Gross BMPR2 gene rearrangements constitute a new cause for primary pulmonary hypertension
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DOI：
10.1097/01.gim.0000156525.09595.e9
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发表时间：
2005-03-01
影响因子：
8.8
通讯作者：
Loyd, JE
中科院分区：
文献类型：
--
作者：
Cogan, JD;Vnencak-Jones, CL;Loyd, JE
目的：据报道，大约50%的家族性原发性肺动脉高压（FPPH）患者存在骨形态发生蛋白受体2型（BMPR2）基因突变。这些突变绝大多数是通过PCR扩增和单个外显子测序确定的。我们研究的目的是确定单独外显子测序未发现的额外BMPR2突变是否可以解释这些阴性病例的重要部分。方法：我们检测了来自12个家族的DNA样本，之前发现BMPR2突变为阴性，以确定任何大的BMPR2基因重排。结果：Southern blot分析发现，在4个（33%）无亲缘关系的类群中存在较大的基因重排。通过逆转录酶PCR （RT-PCR）对其中两种类型的BMPR2转录本进行进一步分析，发现其中一种是杂合的，存在外显子10的重复，另一种是杂合的，存在外显子4至5的缺失。非同源重组被认为是这些大插入/删除的原因。结论：我们的研究结果证明了外显子-外显子测序相关的固有问题，以及其他筛选方法（如Southern blot和RT-PCR）在鉴定BMPR2突变中的重要性。
Purpose: Approximately 50% of patients with familial primary pulmonary hypertension (FPPH) have been reported to have mutations within the bone morphogenic protein receptor type 2 (BMPR2) gene. The vast majority of these mutations were identified by PCR amplification and sequencing of individual exons. The aim of our study was to determine if additional BMPR2 mutations not found by exon sequencing alone could account for a significant portion of these negative cases. Methods: We examined DNA samples from 12 families, previously found to be negative for BMPR2 mutations, to identify any large BMPR2 gene rearrangements. Results: Southern blot analysis found large gene rearrangements in four (33%) unrelated kindreds. Further analysis by reverse transcriptase PCR (RT-PCR) of BMPR2 transcripts from two of these kindreds found one to be heterozygous for a exon 10 duplication and the second to be heterozygous for a deletion of exons 4 to 5. Nonhomologous recombination is believed to be the cause of these large insertions/deletions. Conclusion: Our results demonstrate the inherent problems associated with exon-by-exon sequencing and the importance of other screening methods such as Southern blot and RT-PCR in the identification of BMPR2 mutations.