Molecular Biology Laboratory, Istituto Auxologico Italiano IRCCS, Milan, Italy.
Neuro-Ophthalmology Center and Electrophysiology Laboratory, Department of Ophthalmology, Istituto Auxologico Italiano IRCCS, Milan, Italy.
Exp Eye Res. 2022 Aug;221:109143. doi: 10.1016/j.exer.2022.109143. Epub 2022 Jun 11.
Congenital Stationary Night Blindness type 2 (CSNB2) and Aland island Eye Disease (AIED) associated with CACNA1F mutation demonstrate a significant phenotype overlapping. We report two cases with different clinical presentation carrying two novel mutations in CACNA1F gene. Subjects underwent a complete neurophtahlmological examination associated with structural and electrofunctional insight. Next Generation Sequencing (NGS) analysis of 31 genes previously associated with retinal dystrophy (RD) was performed. Messenger RNAs derived from probands 'peripheral blood samples were analyzed by RT-PCR and cDNA sequencing. The neuro-ophthalmological examinations revealed different clinical, structural and morphological presentations, more severe in patient 1 compared with patient 2. Molecular analysis revealed, that both patients had the hemizygous form of two novel mutations in CACNA1F gene. Patient 1 presented a duplication (c.425dupC) in exon 4, resulting in shifting of the reading frame with the insertion of a premature Stop codon. In Patient 2 variant c.5156G > C localized in the donor's splicing site of exon 43 was identified. Complementary DNA sequencing demonstrated skipping of exon 43 with a deletion of 55 amino acids that causes a frame shift with insertion of a Stop codon. These findings suggest that the effect and the localization of the mutations in the CACNA1F gene can explain different clinical phenotypes. Clinical spectrum is more severe and resembles the AIED phenotype when the mutation affects the first part of the protein, while it is more similar to CSNB2 if the mutation is localized at the end of the protein. Genetic testing results to be an essential tool to provide more accurate diagnosis and prognosis in patients with inherited retinal degenerative disorders and could help, in the future, to develop more specific therapeutic strategies.
先天性静止性夜盲症 2 型 (CSNB2) 和阿兰岛眼病 (AIED) 与 CACNA1F 突变相关,表现出显著的表型重叠。我们报告了两例具有不同临床表现的病例,他们携带 CACNA1F 基因中的两个新突变。受试者接受了完整的神经眼科检查,包括结构和电功能检查。对 31 个先前与视网膜变性 (RD) 相关的基因进行了下一代测序 (NGS) 分析。从先证者的外周血样本中提取信使 RNA,通过 RT-PCR 和 cDNA 测序进行分析。神经眼科检查显示出不同的临床、结构和形态表现,患者 1 比患者 2 更严重。分子分析显示,两名患者均携带 CACNA1F 基因两个新突变的半合子形式。患者 1 在第 4 外显子中存在重复 (c.425dupC),导致阅读框移位并插入一个提前终止密码子。在患者 2 中,鉴定出位于第 43 外显子供体位点的变异 c.5156G > C。互补 DNA 测序显示第 43 外显子缺失,导致 55 个氨基酸缺失,从而导致移码并插入终止密码子。这些发现表明 CACNA1F 基因突变的效应和定位可以解释不同的临床表型。当突变影响蛋白质的前半部分时,临床表型更严重,类似于 AIED 表型,而当突变位于蛋白质的末端时,更类似于 CSNB2。基因检测结果是为遗传性视网膜退行性疾病患者提供更准确诊断和预后的重要工具,并有助于未来开发更具特异性的治疗策略。