Genetic Analysis and Functional Study of a Pedigree With Bruck Syndrome Caused by PLOD2 Variant
BackgroundBruck syndrome (BS) is a rare autosomal recessive inherited osteogenesis imperfecta disease characterized by increased bone fragility and joint contracture. The pathogenic gene of type I BS is FKBPl0, whereas that of type II BS is PLOD2. No significant difference has been found in the clin...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-05-01
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| Series: | Frontiers in Pediatrics |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fped.2022.878172/full |
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| author | Ruo-li Wang Ruo-li Wang Ruo-li Wang Dan-dan Ruan Ya-nan Hu Yu-mian Gan Xin-fu Lin Xin-fu Lin Zhu-ting Fang Zhu-ting Fang Li-sheng Liao Li-sheng Liao Fa-qiang Tang Fa-qiang Tang Wu-bing He Wu-bing He Wu-bing He Jie-wei Luo Jie-wei Luo |
| author_facet | Ruo-li Wang Ruo-li Wang Ruo-li Wang Dan-dan Ruan Ya-nan Hu Yu-mian Gan Xin-fu Lin Xin-fu Lin Zhu-ting Fang Zhu-ting Fang Li-sheng Liao Li-sheng Liao Fa-qiang Tang Fa-qiang Tang Wu-bing He Wu-bing He Wu-bing He Jie-wei Luo Jie-wei Luo |
| author_sort | Ruo-li Wang |
| collection | DOAJ |
| description | BackgroundBruck syndrome (BS) is a rare autosomal recessive inherited osteogenesis imperfecta disease characterized by increased bone fragility and joint contracture. The pathogenic gene of type I BS is FKBPl0, whereas that of type II BS is PLOD2. No significant difference has been found in the clinical phenotype between the two types of BS. In this study, we performed genetic analysis of a BS pedigree caused by PLOD2 variant and studied the corresponding cellular function.MethodsSerum biochemistry, parathyroid hormone (PTH), 25-hydroxyvitamin D [25-(OH) D], osteocalcin, and 24-h urinary calcium levels of a family member with BS was assessed. The genes of the proband were analyzed by second-generation sequencing and exon capture techniques. Sanger sequencing was also performed for the suspected responsible variant of the family member. Wild- and variant-type lentivirus plasmids were constructed by gene cloning and transfected into HEK293T cells. Cell function was verified by real-time quantitative polymerase chain reaction, western blotting, and immunofluorescence detection.ResultsIn this pedigree, the proband was found to have a homozygous variant c.1856G > A (p.Arg619His) in exon 17 of PLOD2 (NM_182943.3). His consanguineous parents and sisters were p.Arg619His heterozygous carriers. The mRNA expression of PLOD2 in the constructed p.Arg619His variant cells was significantly upregulated, while the expression of PLOD2 and collagen I protein in the cell lysate was significantly downregulated. Immunofluorescence revealed that the wild-type PLOD2 was mainly located in the cytoplasm, and the expression of the PLOD2 protein after c.1856G > A variant was significantly downregulated, with almost no expression, aligning with the western blot results. The serum sodium, potassium, calcium, phosphorus, magnesium, alkaline phosphatase, PTH, 25-(OH) D, osteocalcin, and 24 h urinary calcium levels of the proband, his parents, and sisters were normal.ConclusionThrough gene and cell function analyses, PLOD2 Arg619His missense variant was preliminarily confirmed to cause BS by reducing protein expression. |
| first_indexed | 2024-04-13T08:23:53Z |
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| language | English |
| last_indexed | 2024-04-13T08:23:53Z |
| publishDate | 2022-05-01 |
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| series | Frontiers in Pediatrics |
| spelling | doaj.art-b8701ac6f4e6479abef5b44b8644a00e2022-12-22T02:54:33ZengFrontiers Media S.A.Frontiers in Pediatrics2296-23602022-05-011010.3389/fped.2022.878172878172Genetic Analysis and Functional Study of a Pedigree With Bruck Syndrome Caused by PLOD2 VariantRuo-li Wang0Ruo-li Wang1Ruo-li Wang2Dan-dan Ruan3Ya-nan Hu4Yu-mian Gan5Xin-fu Lin6Xin-fu Lin7Zhu-ting Fang8Zhu-ting Fang9Li-sheng Liao10Li-sheng Liao11Fa-qiang Tang12Fa-qiang Tang13Wu-bing He14Wu-bing He15Wu-bing He16Jie-wei Luo17Jie-wei Luo18Shengli Clinical Medical College, Fujian Provincial Hospital, Fujian Medical University, Fuzhou, ChinaDepartment of Emergency, Fujian Provincial Hospital, Fuzhou, ChinaFujian Trauma Medical Center, Fuzhou, ChinaShengli Clinical Medical College, Fujian Provincial Hospital, Fujian Medical University, Fuzhou, ChinaShengli Clinical Medical College, Fujian Provincial Hospital, Fujian Medical University, Fuzhou, ChinaShengli Clinical Medical College, Fujian Provincial Hospital, Fujian Medical University, Fuzhou, ChinaShengli Clinical Medical College, Fujian Provincial Hospital, Fujian Medical University, Fuzhou, ChinaDepartment of Pediatrics, Fujian Provincial Hospital, Fuzhou, ChinaShengli Clinical Medical College, Fujian Provincial Hospital, Fujian Medical University, Fuzhou, ChinaDepartment of Intervention, Fujian Provincial Hospital, Fuzhou, ChinaShengli Clinical Medical College, Fujian Provincial Hospital, Fujian Medical University, Fuzhou, ChinaDepartment of Hematology, Fujian Provincial Hospital, Fuzhou, ChinaShengli Clinical Medical College, Fujian Provincial Hospital, Fujian Medical University, Fuzhou, ChinaDepartment of Orthopedics, Fujian Provincial Hospital, Fuzhou, ChinaShengli Clinical Medical College, Fujian Provincial Hospital, Fujian Medical University, Fuzhou, ChinaDepartment of Emergency, Fujian Provincial Hospital, Fuzhou, ChinaFujian Trauma Medical Center, Fuzhou, ChinaShengli Clinical Medical College, Fujian Provincial Hospital, Fujian Medical University, Fuzhou, ChinaDepartment of Traditional Chinese Medicine, Fujian Provincial Hospital, Fuzhou, ChinaBackgroundBruck syndrome (BS) is a rare autosomal recessive inherited osteogenesis imperfecta disease characterized by increased bone fragility and joint contracture. The pathogenic gene of type I BS is FKBPl0, whereas that of type II BS is PLOD2. No significant difference has been found in the clinical phenotype between the two types of BS. In this study, we performed genetic analysis of a BS pedigree caused by PLOD2 variant and studied the corresponding cellular function.MethodsSerum biochemistry, parathyroid hormone (PTH), 25-hydroxyvitamin D [25-(OH) D], osteocalcin, and 24-h urinary calcium levels of a family member with BS was assessed. The genes of the proband were analyzed by second-generation sequencing and exon capture techniques. Sanger sequencing was also performed for the suspected responsible variant of the family member. Wild- and variant-type lentivirus plasmids were constructed by gene cloning and transfected into HEK293T cells. Cell function was verified by real-time quantitative polymerase chain reaction, western blotting, and immunofluorescence detection.ResultsIn this pedigree, the proband was found to have a homozygous variant c.1856G > A (p.Arg619His) in exon 17 of PLOD2 (NM_182943.3). His consanguineous parents and sisters were p.Arg619His heterozygous carriers. The mRNA expression of PLOD2 in the constructed p.Arg619His variant cells was significantly upregulated, while the expression of PLOD2 and collagen I protein in the cell lysate was significantly downregulated. Immunofluorescence revealed that the wild-type PLOD2 was mainly located in the cytoplasm, and the expression of the PLOD2 protein after c.1856G > A variant was significantly downregulated, with almost no expression, aligning with the western blot results. The serum sodium, potassium, calcium, phosphorus, magnesium, alkaline phosphatase, PTH, 25-(OH) D, osteocalcin, and 24 h urinary calcium levels of the proband, his parents, and sisters were normal.ConclusionThrough gene and cell function analyses, PLOD2 Arg619His missense variant was preliminarily confirmed to cause BS by reducing protein expression.https://www.frontiersin.org/articles/10.3389/fped.2022.878172/fullBruck syndromePLOD2 variantcollagen cross-linkingosteogenesis imperfectajoint contracture |
| spellingShingle | Ruo-li Wang Ruo-li Wang Ruo-li Wang Dan-dan Ruan Ya-nan Hu Yu-mian Gan Xin-fu Lin Xin-fu Lin Zhu-ting Fang Zhu-ting Fang Li-sheng Liao Li-sheng Liao Fa-qiang Tang Fa-qiang Tang Wu-bing He Wu-bing He Wu-bing He Jie-wei Luo Jie-wei Luo Genetic Analysis and Functional Study of a Pedigree With Bruck Syndrome Caused by PLOD2 Variant Frontiers in Pediatrics Bruck syndrome PLOD2 variant collagen cross-linking osteogenesis imperfecta joint contracture |
| title | Genetic Analysis and Functional Study of a Pedigree With Bruck Syndrome Caused by PLOD2 Variant |
| title_full | Genetic Analysis and Functional Study of a Pedigree With Bruck Syndrome Caused by PLOD2 Variant |
| title_fullStr | Genetic Analysis and Functional Study of a Pedigree With Bruck Syndrome Caused by PLOD2 Variant |
| title_full_unstemmed | Genetic Analysis and Functional Study of a Pedigree With Bruck Syndrome Caused by PLOD2 Variant |
| title_short | Genetic Analysis and Functional Study of a Pedigree With Bruck Syndrome Caused by PLOD2 Variant |
| title_sort | genetic analysis and functional study of a pedigree with bruck syndrome caused by plod2 variant |
| topic | Bruck syndrome PLOD2 variant collagen cross-linking osteogenesis imperfecta joint contracture |
| url | https://www.frontiersin.org/articles/10.3389/fped.2022.878172/full |
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