A novel FZD4 mutation p.E160K causes familial exudative vitreoretinopathy_Chinese Journal of Ocular Fundus Diseases

Authors：

XuMin , TianYuanyuan , ChenXue , HuangJiancheng , DingSijia , JiangChao ,  ZhaoChen

Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China;

Corresponding author：

ZhaoChen, Email: dr_zhaochen@163.com

Keywords：

Retinal diseases/genetics; Retinal diseases/etiology; Genes; Mutation; Sequence analysis

DOI：

10.3760/cma.j.issn.1005-1015.2016.06.005

Video：

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Objective To reveal the pathogenic mutation in a three-generation Chinese family with autosomal dominant familial exudative vitreoretinopathy (FEVR). Methods Three patients and a healthy spouse from the index family with FEVR were recruited. The proband was a 5 years old boy. His mother and grandpa were presented with typical FEVR presentations, while his father with normal ocular fundus. DNA was extracted from peripheral blood samples taken from all four participants. All coding and exon-intron boundary regions of five targeted genes, including NDP, FZD4, LRP5, TSPAN12 and ZNF408 were amplified with polymerase chain reaction and sequenced using direct sequencing. In silico analyses were applied to determine the conservation of the mutation site, pathogenic effect and the potential protein crystal structural changes caused by the mutation. Results FZD4 c.478G > A, a susceptible mutation was found after four high frequency mutation sites which MAF values were higher than 0.001 was filtered among 5 single nucleotide variations detected in four participants, leading to the residue 160 changing from glutamate to lysine (p.E160K). Co-segregation analysis between genotypes and phenotypes revealed FZD4 p.E160K as the disease-causing mutation for this family. Conservational analysis suggested that this mutation site was highly conserved among all tested species. Functional analysis predicated that this mutation may be a damaging mutation. Crystal structural analysis also indicated that this mutation could lead to the elimination of the hydrogen bond between residue 160 and asparagine at residue 152, thus altering the tertiary structure of the protein and further impairing the protein function. Conclusion Our study demonstrates FZD4 p.E160K as a novel pathogenic mutation for FEVR.

Citation： XuMin, TianYuanyuan, ChenXue, HuangJiancheng, DingSijia, JiangChao, ZhaoChen. A novel FZD4 mutation p.E160K causes familial exudative vitreoretinopathy. Chinese Journal of Ocular Fundus Diseases, 2016, 32(6): 582-586. doi: 10.3760/cma.j.issn.1005-1015.2016.06.005 Copy

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2.	Li Y, Muller B, Fuhrmann C, et al. The autosomal dominant familial exudative vitreoretinopathy locus maps on 11q and is closely linked to D11S533[J]. Am J Hum Genet, 1992, 51(4):749-754.
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11.	陈雪, 高翔, 赵堪兴, 等.目标序列捕获测序发现视网膜色素变性的热点突变SNRNP200 p.S1087L及其新临床表型[J].中华眼科杂志, 2013, 49(12):1104-1110. DOI:10.3760/cma.j.issn.0412-4081.2013.12.009.Chen X, Gao X, Zhao KX, et al.Targeted sequencing identifies a hotspot mutation SNRNP200 p.S1087L correlates with novel phenotypes in retinitis pigmentosa[J]. Chin J Ophthalmol, 2013, 49(12):1104-1110. DOI:10.3760/cma.j.issn. 0412-4081. 2013.12.009.
12.	Adzhubei IA, Schmidt S, Peshkin L, et al. A method and server for predicting damaging missense mutations[J]. Nat Methods, 2010, 7(4):248-249.DOI:10.1038/nmeth0410-248.
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14.	Arnold K, Bordoli L, Kopp J, et al. The SWISS-MODEL workspace:a web-based environment for protein structure homology modelling[J]. Bioinformatics, 2006, 22(2):195-201.DOI:10.1093/bioinformatics/bti770.
15.	Kiefer F, Arnold K, Kunzli M, et al. The SWISS-MODEL Repository and associated resources[J]. Nucleic Acids Res, 2009, 37:387-392. DOI:10.1093/nar/gkn750.
16.	Milhem RM, Ben-Salem S, Al-Gazali L, et al.Identification of the cellular mechanisms that modulate trafficking of frizzled family receptor 4(FZD4) missense mutants associated with familial exudative vitreoretinopathy[J].Invest Ophthalmol Vis Sci, 2014, 55(6):3423-3431.DOI:10.1167/iovs.14-13885.
17.	Clevers H, Nusse R. Wnt/beta-catenin signaling and disease[J]. Cell, 2012, 149(6):1192-1205.DOI:10.1016/j.cell.2012.05.012.
18.	Kondo H, Hayashi H, Oshima K, et al. Frizzled 4 gene (FZD4) mutations in patients with familial exudative vitreoretinopathy with variable expressivity[J]. Br J Ophthalmol, 2003, 87(10):1291-1295.
19.	Fei P, Zhu X, Jiang Z, et al. Identification and functional analysis of novel FZD4 mutations in Han Chinese with familial exudative vitreoretinopathy[J]. Sci Rep, 2015, 5:16120. DOI:10.1038/srep16120.
20.	Qin M, Hayashi H, Oshima K, et al. Complexity of the genotype-phenotype correlation in familial exudative vitreoretinopathy with mutations in the LRP5 and/or FZD4 genes[J]. Hum Mutat, 2005, 26(2):104-112.DOI:10.1002/humu.20191.
21.	李忆安, 张琦, 彭婕, 等.卷曲蛋白4联合低密度脂蛋白相关蛋白5基因突变的家族性渗出性玻璃体视网膜病变二例[J].中华眼底病杂志, 2016, 32(1):79-81.DOI:10.3760/cma.j.issn.1005-1015.2016.01.019.Li YA, Zhang Q, Peng J, et al.Two cases of frizzled 4 gene (FZD4) mutation joint with low density lipoprotein associated protein 5 gene mutation in familial exudative vitreoretinopathy retinopathy[J].Chin J Ocul Fundus Dis, 2016, 32(1):79-81.DOI:10.3760/cma.j.issn.1005-1015.2016.01.019.
22.	Qin M, Kondo H, Tahira T, et al. Moderate reduction of Norrin signaling activity associated with the causative missense mutations identified in patients with familial exudative vitreoretinopathy[J]. Hum Genet, 2008, 122(6):615-623.

1. Chen ZY, Battinelli EM, Fielder A, et al. A mutation in the Norrie disease gene (NDP) associated with X-linked familial exudative vitreoretinopathy[J]. Nat Genet, 1993, 5(2):180-183.DOI:10.1038/nq1093-180.
2. Li Y, Muller B, Fuhrmann C, et al. The autosomal dominant familial exudative vitreoretinopathy locus maps on 11q and is closely linked to D11S533[J]. Am J Hum Genet, 1992, 51(4):749-754.
3. Robitaille J, Macdonald ML, Kaykas A, et al. Mutant frizzled-4 disrupts retinal angiogenesis in familial exudative vitreoretinopathy[J]. Nat Genet, 2002, 32(2):326-330.DOI:10.1038/nq957.
4. Toomes C, Bottomley HM, Jackson RM, et al. Mutations in LRP5 or FZD4 underlie the common familial exudative vitreoretinopathy locus on chromosome 11q[J]. Am J Hum Genet, 2004, 74(4):721-730.DOI:10.1086/383202.
5. Nikopoulos K, Gilissen C, Hoischen A, et al. Next-generation sequencing of a 40 Mb linkage interval reveals TSPAN12 mutations in patients with familial exudative vitreoretinopathy[J]. Am J Hum Genet, 2010, 86(2):240-247.DOI:10.1016/j.ajhg.2009.12.016.
6. Collin RW, Nikopoulos K, Dona M, et al. ZNF408 is mutated in familial exudative vitreoretinopathy and is crucial for the development of zebrafish retinal vasculature[J]. Proc Natl Acad Sci USA, 2013, 110(24):9856-9861.DOI:10.1073/pnas.1220864110.
7. MacDonald BT, Tamai K, He X.Wnt/beta-catenin signaling:components, mechanisms, and diseases[J].Dev Cell, 2009, 17(1):9-26. DOI:10.1016/j.Devcel. 2009.06.016.DOI:10.1016/j.devcel.2009.06.016.
8. Seo SH, Yu YS, Park SW, et al. Molecular characterization of FZD4, LRP5, and TSPAN12 in familial exudative vitreoretinopathy[J]. Invest Ophthalmol Vis Sci, 2015, 56(9):5143-5151.DOI:10.1167/iovs.14-15680.
9. Nikopoulos K, Venselaar H, Collin RW, et al. Overview of the mutation spectrum in familial exudative vitreoretinopathy and Norrie disease with identification of 21 novel variants in FZD4, LRP5, and NDP[J]. Hum Mutat, 2010, 31(6):656-666.DOI:10.1002/humu.21250.
10. 刘巾男, 胥婷婷, 辛梅, 等.家族性渗出性玻璃体视网膜病变的眼底分析[J].中国循证医学杂志, 2014, 14(6):668-670.DOI:10.7507/1672-2531.20140113.Liu JN, Xu TT, Xin M, et al.Fundus analysis of familial exudative vitreoretinopathy[J].Chin J Evid-Based Med, 2014, 14(6):668-670.DOI:10.7507/1672-2531.20140113.
11. 陈雪, 高翔, 赵堪兴, 等.目标序列捕获测序发现视网膜色素变性的热点突变SNRNP200 p.S1087L及其新临床表型[J].中华眼科杂志, 2013, 49(12):1104-1110. DOI:10.3760/cma.j.issn.0412-4081.2013.12.009.Chen X, Gao X, Zhao KX, et al.Targeted sequencing identifies a hotspot mutation SNRNP200 p.S1087L correlates with novel phenotypes in retinitis pigmentosa[J]. Chin J Ophthalmol, 2013, 49(12):1104-1110. DOI:10.3760/cma.j.issn. 0412-4081. 2013.12.009.
12. Adzhubei IA, Schmidt S, Peshkin L, et al. A method and server for predicting damaging missense mutations[J]. Nat Methods, 2010, 7(4):248-249.DOI:10.1038/nmeth0410-248.
13. Tavtigian SV, Deffenbaugh AM, Yin L, et al. Comprehensive statistical study of 452 BRCA1 missense substitutions with classification of eight recurrent substitutions as neutral[J]. J Med Genet, 2006, 43(4):295-305.DOI:10.1136/jmg. 2005.033878.
14. Arnold K, Bordoli L, Kopp J, et al. The SWISS-MODEL workspace:a web-based environment for protein structure homology modelling[J]. Bioinformatics, 2006, 22(2):195-201.DOI:10.1093/bioinformatics/bti770.
15. Kiefer F, Arnold K, Kunzli M, et al. The SWISS-MODEL Repository and associated resources[J]. Nucleic Acids Res, 2009, 37:387-392. DOI:10.1093/nar/gkn750.
16. Milhem RM, Ben-Salem S, Al-Gazali L, et al.Identification of the cellular mechanisms that modulate trafficking of frizzled family receptor 4(FZD4) missense mutants associated with familial exudative vitreoretinopathy[J].Invest Ophthalmol Vis Sci, 2014, 55(6):3423-3431.DOI:10.1167/iovs.14-13885.
17. Clevers H, Nusse R. Wnt/beta-catenin signaling and disease[J]. Cell, 2012, 149(6):1192-1205.DOI:10.1016/j.cell.2012.05.012.
18. Kondo H, Hayashi H, Oshima K, et al. Frizzled 4 gene (FZD4) mutations in patients with familial exudative vitreoretinopathy with variable expressivity[J]. Br J Ophthalmol, 2003, 87(10):1291-1295.
19. Fei P, Zhu X, Jiang Z, et al. Identification and functional analysis of novel FZD4 mutations in Han Chinese with familial exudative vitreoretinopathy[J]. Sci Rep, 2015, 5:16120. DOI:10.1038/srep16120.
20. Qin M, Hayashi H, Oshima K, et al. Complexity of the genotype-phenotype correlation in familial exudative vitreoretinopathy with mutations in the LRP5 and/or FZD4 genes[J]. Hum Mutat, 2005, 26(2):104-112.DOI:10.1002/humu.20191.
21. 李忆安, 张琦, 彭婕, 等.卷曲蛋白4联合低密度脂蛋白相关蛋白5基因突变的家族性渗出性玻璃体视网膜病变二例[J].中华眼底病杂志, 2016, 32(1):79-81.DOI:10.3760/cma.j.issn.1005-1015.2016.01.019.Li YA, Zhang Q, Peng J, et al.Two cases of frizzled 4 gene (FZD4) mutation joint with low density lipoprotein associated protein 5 gene mutation in familial exudative vitreoretinopathy retinopathy[J].Chin J Ocul Fundus Dis, 2016, 32(1):79-81.DOI:10.3760/cma.j.issn.1005-1015.2016.01.019.
22. Qin M, Kondo H, Tahira T, et al. Moderate reduction of Norrin signaling activity associated with the causative missense mutations identified in patients with familial exudative vitreoretinopathy[J]. Hum Genet, 2008, 122(6):615-623.

Chinese Journal of Ocular Fundus Diseases

A novel FZD4 mutation p.E160K causes familial exudative vitreoretinopathy

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