Hudson Gavin, Yu-Wai-Man Patrick, Griffiths Phillip G, Caporali Leonardo, Salomao Solange S, Berezovsky Adriana, Carelli Valerio, Zeviani Massimo, Chinnery Patrick F
Institute for Human Genetics, Newcastle University, Newcastle upon Tyne, UK.
Mol Vis. 2010 Dec 15;16:2760-4.
Leber hereditary optic neuropathy (LHON) is a common cause of inherited blindness, primarily due to one of three mitochondrial DNA (mtDNA) mutations. These mtDNA pathogenic mutations have variable clinical penetrance. Recent linkage evidence raised the possibility that the nuclear gene optic atrophy 1 (OPA1) determines whether mtDNA mutation carriers develop blindness. To validate these findings we studied OPA1 in three independent LHON cohorts: sequencing the gene in discordant male sib pairs, carrying out a family-based association study of common functional genetic variants, and carrying out a population-based association study of the same genetic variants.
We tested 3 hypothesis in three separate study groups. Study group 1: Direct sequencing of OPA1 coding regions was performed using sequencing methodologies (Applied Biosystems, Foster City, CA). Chromatograms were compared with the GenBank reference sequence NM_015560.1. Splice-site prediction was performed using GeneSplicer. Study group 2: Genotyping for rs166850 and rs10451941 was performed by restriction fragment length polymorphism (RFLP) analysis with specific primers for both genotypes, using The restriction enzymes RsaI and FspBI to discriminate genotypes. Study group 3: Genotyping for rs166850 and rs10451941 was performed by primer extension of allele-specific extensions products by matrix-associated laser desorption/ionisation time-of-flight (MALDI-TOF, Seqeunom, San Diego, CA) mass spectrometry. Allele and genotype frequencies were compared using Pearson's chi-square test. Multiple logistic regression was performed to look for interactions between the variables. All analyses were performed using SPSS software version 17.0 (SPSS Inc.).
In all three groups we were unable to find an association between OPA1 genetic variation and visual failure in LHON mtDNA mutation carriers.
Our findings suggest that genetic variation in OPA1 is unlikely to make a major contribution to the risk of blindness in LHON mutation carriers.
Leber遗传性视神经病变（LHON）是遗传性失明的常见病因，主要由三种线粒体DNA（mtDNA）突变之一引起。这些mtDNA致病突变具有不同的临床外显率。最近的连锁证据提示核基因视神经萎缩1（OPA1）可能决定mtDNA突变携带者是否会发展为失明。为验证这些发现，我们在三个独立的LHON队列中研究了OPA1：对不一致的男性同胞对进行该基因测序，对常见功能基因变异进行基于家系的关联研究，并对相同的基因变异进行基于人群的关联研究。
我们在三个独立的研究组中检验了3个假设。研究组1：使用测序方法（Applied Biosystems，加利福尼亚州福斯特城）对OPA1编码区进行直接测序。将色谱图与GenBank参考序列NM_015560.1进行比较。使用GeneSplicer进行剪接位点预测。研究组2：通过限制性片段长度多态性（RFLP）分析对rs166850和rs10451941进行基因分型，针对两种基因型使用特异性引物，使用限制性内切酶RsaI和FspBI区分基因型。研究组3：通过基质辅助激光解吸/电离飞行时间（MALDI-TOF，Sequenom，加利福尼亚州圣地亚哥）质谱对等位基因特异性延伸产物进行引物延伸，对rs166850和rs10451941进行基因分型。使用Pearson卡方检验比较等位基因和基因型频率。进行多因素逻辑回归以寻找变量之间的相互作用。所有分析均使用SPSS软件17.0版（SPSS公司）进行。
在所有三个组中，我们均未发现OPA1基因变异与LHON mtDNA突变携带者的视力丧失之间存在关联。
我们的研究结果表明，OPA1基因变异不太可能对LHON突变携带者的失明风险产生重大影响。
