IFT88 mutations identified in individuals with non-syndromic recessive retinal degeneration result in abnormal ciliogenesis
IFT88 mutations identified in individuals with non-syndromic recessive retinal degeneration result in abnormal ciliogenesis
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DOI：
10.1007/s00439-018-1897-9
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发表时间：
2018-07-01
期刊：
影响因子：
5.3
通讯作者：
Ayyagari, Radha
中科院分区：
文献类型：
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作者：
Chekuri, Anil;Guru, Aditya A.;Ayyagari, Radha
通过全基因组测序（WGS）来鉴定导致白种人家族遗传性视网膜变性（IRD）的变异。对选定的具有致病潜力的罕见变异进行分离分析，确定了与 IRD 分离的鞭毛内转运蛋白 88 (IFT88) 基因中的一组复合杂合变化 p.Arg266*:c.796C > T 和 p.Ala568Thr:c.1702G > A。通过瞬时转染在 mIMDC3 细胞中以及通过使用 CRISPR-cas9 系统引入突变在 HeLa 细胞中表达带有 p.Arg266* 和 p.Ala568Thr 突变的 IFT88，表明这两种突变都会导致异常纤毛结构的形成。通过 CRISPR-Cas9 基因组编辑在 HeLa 细胞中以纯合状态引入 IFT88 p.Arg266* 变体，结果表明突变转录本经历无义介导的衰变，导致 IFT88 转录本显着缺失。此外，在这些细胞中观察到纤毛发生异常。这些观察结果表明，本研究中观察到的 IFT88 等位基因的罕见且独特的组合提供了对 IFT88 在人类中的生理作用以及 IRD 谱系中视网膜病理学潜在机制的深入了解。
Whole genome sequencing (WGS) was performed to identify the variants responsible for inherited retinal degeneration (IRD) in a Caucasian family. Segregation analysis of selected rare variants with pathogenic potential identified a set of compound heterozygous changes p.Arg266*:c.796C > T and p.Ala568Thr:c.1702G > A in the intraflagellar transport protein-88 (IFT88) gene segregating with IRD. Expression of IFT88 with the p.Arg266* and p.Ala568Thr mutations in mIMDC3 cells by transient transfection and in HeLa cells by introducing the mutations using CRISPR-cas9 system suggested that both mutations result in the formation of abnormal ciliary structures. The introduction of the IFT88 p.Arg266* variant in the homozygous state in HeLa cells by CRISPR-Cas9 genome-editing revealed that the mutant transcript undergoes nonsense-mediated decay leading to a significant depletion of IFT88 transcript. Additionally, abnormal ciliogenesis was observed in these cells. These observations suggest that the rare and unique combination of IFT88 alleles observed in this study provide insight into the physiological role of IFT88 in humans and the likely mechanism underlying retinal pathology in the pedigree with IRD.