Interpreting Variants of Uncertain Significance in PCD: Abnormal Splicing Caused by a Missense Variant of DNAAF3
ABSTRACT Background Primary ciliary dyskinesia (PCD) is a rare autosomal recessive disorder characterized by dysfunction of motile cilia. While approximately 50 genes have been identified, around 25% of PCD patients remain genetically unexplained; elucidating the pathogenicity of specific variants r...
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Wiley
2025-01-01
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| Series: | Molecular Genetics & Genomic Medicine |
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| Online Access: | https://doi.org/10.1002/mgg3.70036 |
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| author | Haixia Zheng Chongsheng Cheng Miao He Wangji Zhou Yixuan Li Jinrong Dai Ting Zhang Kai‐Feng Xu Xue Zhang Xinlun Tian Yaping Liu |
| author_facet | Haixia Zheng Chongsheng Cheng Miao He Wangji Zhou Yixuan Li Jinrong Dai Ting Zhang Kai‐Feng Xu Xue Zhang Xinlun Tian Yaping Liu |
| author_sort | Haixia Zheng |
| collection | DOAJ |
| description | ABSTRACT Background Primary ciliary dyskinesia (PCD) is a rare autosomal recessive disorder characterized by dysfunction of motile cilia. While approximately 50 genes have been identified, around 25% of PCD patients remain genetically unexplained; elucidating the pathogenicity of specific variants remains a challenge. Methods Whole exome sequencing (WES) and Sanger sequencing were conducted to identify potential pathogenic variants of PCD. Minigene assays were performed to evaluate the pathogenicity of variants. Transmission electron microscopy (TEM) and high‐speed video analysis (HSVA) were conducted to analyze the function of cilia in respiratory epithelial cells. Results We identified two variants of DNAAF3: c.557G>A, p.G186E in exon 5, and c.1364G>A, p.G455D at the terminal nucleotide of exon 10 in a 16‐year‐old male patient. Through a minigene assay, we demonstrated that the c.1364G>A variant led to a four‐nucleotide skipping. The cilia in epithelial ciliary cells of the proband were almost immotile. The absence of outer dynein arms and inner dynein arms was also observed. Conclusions Our study identified two compound heterozygous variants of DNAAF3, a pathogenic gene for PCD, and proved that a novel missense variant c.1364G>A affects splicing. Our findings not only expanded the spectrum of mutations in the DNAAF3 gene but also highlighted the importance of investigating variants of uncertain significance (VUS) for comprehensive genetic diagnoses. |
| format | Article |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-01-01 |
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| series | Molecular Genetics & Genomic Medicine |
| spelling | doaj-art-4b174286df0e492eb608b2a7a9ed2f1c2025-01-24T08:16:42ZengWileyMolecular Genetics & Genomic Medicine2324-92692025-01-01131n/an/a10.1002/mgg3.70036Interpreting Variants of Uncertain Significance in PCD: Abnormal Splicing Caused by a Missense Variant of DNAAF3Haixia Zheng0Chongsheng Cheng1Miao He2Wangji Zhou3Yixuan Li4Jinrong Dai5Ting Zhang6Kai‐Feng Xu7Xue Zhang8Xinlun Tian9Yaping Liu10McKusick‐Zhang Center for Genetic Medicine, State Key Laboratory for Complex Severe and Rare Diseases, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences School of Basic Medicine, Peking Union Medical College Beijing ChinaDepartment of Pulmonary and Critical Care Medicine, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital Chinese Academy of Medical Sciences & Peking Union Medical College Beijing ChinaMcKusick‐Zhang Center for Genetic Medicine, State Key Laboratory for Complex Severe and Rare Diseases, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences School of Basic Medicine, Peking Union Medical College Beijing ChinaDepartment of Pulmonary and Critical Care Medicine, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital Chinese Academy of Medical Sciences & Peking Union Medical College Beijing ChinaMcKusick‐Zhang Center for Genetic Medicine, State Key Laboratory for Complex Severe and Rare Diseases, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences School of Basic Medicine, Peking Union Medical College Beijing ChinaDepartment of Pulmonary and Critical Care Medicine, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital Chinese Academy of Medical Sciences & Peking Union Medical College Beijing ChinaDepartment of Pulmonary and Critical Care Medicine, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital Chinese Academy of Medical Sciences & Peking Union Medical College Beijing ChinaDepartment of Pulmonary and Critical Care Medicine, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital Chinese Academy of Medical Sciences & Peking Union Medical College Beijing ChinaMcKusick‐Zhang Center for Genetic Medicine, State Key Laboratory for Complex Severe and Rare Diseases, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences School of Basic Medicine, Peking Union Medical College Beijing ChinaDepartment of Pulmonary and Critical Care Medicine, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital Chinese Academy of Medical Sciences & Peking Union Medical College Beijing ChinaThe State Key Laboratory for Complex Severe and Rare Diseases, the State Key Sci‐Tech Infrastructure for Translational Medicine Peking Union Medical College Hospital Beijing ChinaABSTRACT Background Primary ciliary dyskinesia (PCD) is a rare autosomal recessive disorder characterized by dysfunction of motile cilia. While approximately 50 genes have been identified, around 25% of PCD patients remain genetically unexplained; elucidating the pathogenicity of specific variants remains a challenge. Methods Whole exome sequencing (WES) and Sanger sequencing were conducted to identify potential pathogenic variants of PCD. Minigene assays were performed to evaluate the pathogenicity of variants. Transmission electron microscopy (TEM) and high‐speed video analysis (HSVA) were conducted to analyze the function of cilia in respiratory epithelial cells. Results We identified two variants of DNAAF3: c.557G>A, p.G186E in exon 5, and c.1364G>A, p.G455D at the terminal nucleotide of exon 10 in a 16‐year‐old male patient. Through a minigene assay, we demonstrated that the c.1364G>A variant led to a four‐nucleotide skipping. The cilia in epithelial ciliary cells of the proband were almost immotile. The absence of outer dynein arms and inner dynein arms was also observed. Conclusions Our study identified two compound heterozygous variants of DNAAF3, a pathogenic gene for PCD, and proved that a novel missense variant c.1364G>A affects splicing. Our findings not only expanded the spectrum of mutations in the DNAAF3 gene but also highlighted the importance of investigating variants of uncertain significance (VUS) for comprehensive genetic diagnoses.https://doi.org/10.1002/mgg3.70036DNAAF3minigene assaymolecular diagnosisprimary ciliary dyskinesia (PCD)variants of uncertain significance (VUS) |
| spellingShingle | Haixia Zheng Chongsheng Cheng Miao He Wangji Zhou Yixuan Li Jinrong Dai Ting Zhang Kai‐Feng Xu Xue Zhang Xinlun Tian Yaping Liu Interpreting Variants of Uncertain Significance in PCD: Abnormal Splicing Caused by a Missense Variant of DNAAF3 Molecular Genetics & Genomic Medicine DNAAF3 minigene assay molecular diagnosis primary ciliary dyskinesia (PCD) variants of uncertain significance (VUS) |
| title | Interpreting Variants of Uncertain Significance in PCD: Abnormal Splicing Caused by a Missense Variant of DNAAF3 |
| title_full | Interpreting Variants of Uncertain Significance in PCD: Abnormal Splicing Caused by a Missense Variant of DNAAF3 |
| title_fullStr | Interpreting Variants of Uncertain Significance in PCD: Abnormal Splicing Caused by a Missense Variant of DNAAF3 |
| title_full_unstemmed | Interpreting Variants of Uncertain Significance in PCD: Abnormal Splicing Caused by a Missense Variant of DNAAF3 |
| title_short | Interpreting Variants of Uncertain Significance in PCD: Abnormal Splicing Caused by a Missense Variant of DNAAF3 |
| title_sort | interpreting variants of uncertain significance in pcd abnormal splicing caused by a missense variant of dnaaf3 |
| topic | DNAAF3 minigene assay molecular diagnosis primary ciliary dyskinesia (PCD) variants of uncertain significance (VUS) |
| url | https://doi.org/10.1002/mgg3.70036 |
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