Shimizu Norihiro, Mashimo Yoichi, Yokouchi Hirotaka, Nishio Yosuke, Sawai Setsu, Ichikawa Tomohiko, Ogi Tomoo, Baba Takayuki, Onouchi Yoshihiro
Department of Ophthalmology and Visual Science, Chiba University Graduate School of Medicine, Chiba, Japan.
Maebara Shimizu Eye Clinic, Funabashi, Japan.
J Hum Genet. 2025 Apr;70(4):199-205. doi: 10.1038/s10038-025-01318-0. Epub 2025 Feb 13.
Mutations in fibrillin-1 (FBN1) cause various clinical conditions, such as Marfan syndrome (MFS). However, the genotype-phenotype relationships underlying MFS and other conditions relevant to FBN1 mutations have not been fully elucidated. We performed whole-exome sequencing on three participants, including an affected mother-daughter pair, in a three-generation Japanese family with isolated ectopia lentis (IEL). The sequencing identified a novel single-nucleotide variant (c.1327+3A>C) in intron 11 of FBN1 that was shared between the two patients. We confirmed the co-segregation of the variant with IEL in two additional affected relatives in the family. The Combined Annotation-Dependent Depletion score of the variant was 26.1, which was indicated by SpliceAI to influence splicing, with a score of 0.93. Reverse transcription-polymerase chain reaction (RT-PCR) of mRNAs isolated from peripheral blood mononuclear cells revealed aberrant bands in all four affected individuals. Subsequent sequencing revealed that these bands originated from FBN1 transcripts lacking exon 11. The causality of the variant in the skipping of exon 11, which results in an in-frame deletion of 60 amino acids corresponding to the "hinge" region of FBN1 protein, was confirmed in a minigene experiment. Interestingly, the same result was observed for a minigene for c.1327+1G>A, a variant previously identified in two unrelated EL families without MFS manifestations. These results suggest that the c.1327+3A>C mutation in FBN1 likely leads to IEL. The findings expand our knowledge of FBN1 and provide insights into FBN1-related diseases.
原纤蛋白-1(FBN1)的突变会导致多种临床病症,如马凡综合征(MFS)。然而,MFS以及与FBN1突变相关的其他病症的基因型-表型关系尚未完全阐明。我们对一个三代日本孤立性晶状体异位(IEL)家族中的三名参与者进行了全外显子组测序,其中包括一对患病的母女。测序在FBN1的第11内含子中鉴定出一个新的单核苷酸变异(c.1327+3A>C),该变异在两名患者中共有。我们在该家族另外两名患病亲属中证实了该变异与IEL的共分离。该变异的联合注释依赖损耗分数为26.1,SpliceAI表明其会影响剪接,分数为0.93。从外周血单个核细胞分离的mRNA进行逆转录-聚合酶链反应(RT-PCR)显示,所有四名患病个体均出现异常条带。随后的测序表明,这些条带源自缺少外显子11的FBN1转录本。在一个小基因实验中证实了该变异导致外显子11跳跃的因果关系,这导致FBN1蛋白“铰链”区域对应的60个氨基酸的框内缺失。有趣的是,对于c.1327+1G>A的小基因也观察到了相同的结果,该变异先前在两个无MFS表现的无关EL家族中被鉴定出来。这些结果表明,FBN1中的c.1327+3A>C突变可能导致IEL。这些发现扩展了我们对FBN1的认识,并为FBN1相关疾病提供了见解。