Novel mutations in COL4A3, COL4A4, and COL4A5 in Chinese patients with Alport Syndrome.
Alport syndrome (AS) is a clinically and genetically heterogeneous, progressive nephropathy caused by mutations in COL4A3, COL4A4, and COL4A5, which encode type IV collagen. The large sizes of these genes and the absence of mutation hot spots have complicated mutational analysis by routine polymeras...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2017-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC5436713?pdf=render |
| _version_ | 1819109278705778688 |
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| author | Jian-Hong Liu Xiu-Xiu Wei Ang Li Ying-Xia Cui Xin-Yi Xia Wei-Song Qin Ming-Chao Zhang Er-Zhi Gao Jun Sun Chun-Lin Gao Feng-Xia Liu Qiu-Yue Wu Wei-Wei Li Asan Zhi-Hong Liu Xiao-Jun Li |
| author_facet | Jian-Hong Liu Xiu-Xiu Wei Ang Li Ying-Xia Cui Xin-Yi Xia Wei-Song Qin Ming-Chao Zhang Er-Zhi Gao Jun Sun Chun-Lin Gao Feng-Xia Liu Qiu-Yue Wu Wei-Wei Li Asan Zhi-Hong Liu Xiao-Jun Li |
| author_sort | Jian-Hong Liu |
| collection | DOAJ |
| description | Alport syndrome (AS) is a clinically and genetically heterogeneous, progressive nephropathy caused by mutations in COL4A3, COL4A4, and COL4A5, which encode type IV collagen. The large sizes of these genes and the absence of mutation hot spots have complicated mutational analysis by routine polymerase chain reaction (PCR)-based approaches. Here, in order to design a rapid and effective method for the genetic diagnosis of AS, we developed a strategy by utilizing targeted capture associated with next-generation sequencing (NGS) to analyze COL4A3, COL4A4, and COL4A5 simultaneously in 20 AS patients. All the coding exons and flanking sequences of COL4A3, COL4A4, and COL4A5 from the probands were captured followed by HiSeq 2500 sequencing. Candidate mutations were validated by classic Sanger sequencing and quantitative (q)PCR. Sixteen patients (16/20, 75%) showed X-linked inheritance, and four patients (4/20, 20%) showed autosomal recessive inheritance. None of the individuals had autosomal-dominant AS. Fifteen novel mutations, 6 known mutations, and 2 novel fragment deletions were detected by targeted capture and NGS. Of these novel mutations, 12, 3, and 2 mutations were detected in COL4A5, COL4A4, and COL4A3, respectively. A comparison of the clinical manifestations caused by different types of mutations in COL4A5 suggested that nonsense mutations and glycine substitution by an acidic amino acid are more severe than the other missense mutations. Pathogenic mutations were detected in 20 patients. These novel mutations can expand the genotypic spectrum of AS. Our results demonstrated that targeted capture and NGS technology are effective in the genetic diagnosis of AS. |
| first_indexed | 2024-12-22T03:23:17Z |
| format | Article |
| id | doaj.art-5d8fd1cbfd01476ca700cca2280e75f4 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-22T03:23:17Z |
| publishDate | 2017-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj.art-5d8fd1cbfd01476ca700cca2280e75f42022-12-21T18:40:40ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01125e017768510.1371/journal.pone.0177685Novel mutations in COL4A3, COL4A4, and COL4A5 in Chinese patients with Alport Syndrome.Jian-Hong LiuXiu-Xiu WeiAng LiYing-Xia CuiXin-Yi XiaWei-Song QinMing-Chao ZhangEr-Zhi GaoJun SunChun-Lin GaoFeng-Xia LiuQiu-Yue WuWei-Wei LiAsanZhi-Hong LiuXiao-Jun LiAlport syndrome (AS) is a clinically and genetically heterogeneous, progressive nephropathy caused by mutations in COL4A3, COL4A4, and COL4A5, which encode type IV collagen. The large sizes of these genes and the absence of mutation hot spots have complicated mutational analysis by routine polymerase chain reaction (PCR)-based approaches. Here, in order to design a rapid and effective method for the genetic diagnosis of AS, we developed a strategy by utilizing targeted capture associated with next-generation sequencing (NGS) to analyze COL4A3, COL4A4, and COL4A5 simultaneously in 20 AS patients. All the coding exons and flanking sequences of COL4A3, COL4A4, and COL4A5 from the probands were captured followed by HiSeq 2500 sequencing. Candidate mutations were validated by classic Sanger sequencing and quantitative (q)PCR. Sixteen patients (16/20, 75%) showed X-linked inheritance, and four patients (4/20, 20%) showed autosomal recessive inheritance. None of the individuals had autosomal-dominant AS. Fifteen novel mutations, 6 known mutations, and 2 novel fragment deletions were detected by targeted capture and NGS. Of these novel mutations, 12, 3, and 2 mutations were detected in COL4A5, COL4A4, and COL4A3, respectively. A comparison of the clinical manifestations caused by different types of mutations in COL4A5 suggested that nonsense mutations and glycine substitution by an acidic amino acid are more severe than the other missense mutations. Pathogenic mutations were detected in 20 patients. These novel mutations can expand the genotypic spectrum of AS. Our results demonstrated that targeted capture and NGS technology are effective in the genetic diagnosis of AS.http://europepmc.org/articles/PMC5436713?pdf=render |
| spellingShingle | Jian-Hong Liu Xiu-Xiu Wei Ang Li Ying-Xia Cui Xin-Yi Xia Wei-Song Qin Ming-Chao Zhang Er-Zhi Gao Jun Sun Chun-Lin Gao Feng-Xia Liu Qiu-Yue Wu Wei-Wei Li Asan Zhi-Hong Liu Xiao-Jun Li Novel mutations in COL4A3, COL4A4, and COL4A5 in Chinese patients with Alport Syndrome. PLoS ONE |
| title | Novel mutations in COL4A3, COL4A4, and COL4A5 in Chinese patients with Alport Syndrome. |
| title_full | Novel mutations in COL4A3, COL4A4, and COL4A5 in Chinese patients with Alport Syndrome. |
| title_fullStr | Novel mutations in COL4A3, COL4A4, and COL4A5 in Chinese patients with Alport Syndrome. |
| title_full_unstemmed | Novel mutations in COL4A3, COL4A4, and COL4A5 in Chinese patients with Alport Syndrome. |
| title_short | Novel mutations in COL4A3, COL4A4, and COL4A5 in Chinese patients with Alport Syndrome. |
| title_sort | novel mutations in col4a3 col4a4 and col4a5 in chinese patients with alport syndrome |
| url | http://europepmc.org/articles/PMC5436713?pdf=render |
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