PDF(Pubmed)
Abstract：
BACKGROUND: To report newly found TSPAN12 mutations with a unique form of familial exudative vitreoretinopathy (FEVR) and find out the possible mechanism of a repeated novel intronic variant in TSPAN12 led to FEVR.
RESULTS: Nine TSPAN12 mutations with a unique form of FEVR were detected by panel-based NGS. MINI-Gene assay showed two splicing modes of mRNA that process two different bands A and B, and mutant-type shows replacement with the splicing mode of Exon11 hopping. Construction of wild-type and mutant TSPAN12 vector showed the appearance of premature termination codons (PTC). In vitro expression detection showed significant down-regulated expression level of TSPAN12 mRNAs and proteins in cells transfected with mutant vectors compared with in wild-type group. On the contrary, translation inhibitor CHX and small interfering RNA of UPF1 (si-UPF1) significantly increased mRNA or protein expression of TSPAN12 in cells transfected with the mutant vectors.
CONCLUSIONS: Nine mutations in TSPAN12 gene are reported in 9 FEVR patients with a unique series of ocular abnormalities. The three novel TSPAN12 mutations trigger NMD would cause the decrease of TSPAN12 proteins that participate in biosynthesis and assembly of microfibers, which might lead to FEVR, and suggest that intronic sequence analysis might be a vital tool for genetic counseling and prenatal diagnoses.
RESULTS: Nine TSPAN12 mutations with a unique form of FEVR were detected by panel-based NGS. MINI-Gene assay showed two splicing modes of mRNA that process two different bands A and B, and mutant-type shows replacement with the splicing mode of Exon11 hopping. Construction of wild-type and mutant TSPAN12 vector showed the appearance of premature termination codons (PTC). In vitro expression detection showed significant down-regulated expression level of TSPAN12 mRNAs and proteins in cells transfected with mutant vectors compared with in wild-type group. On the contrary, translation inhibitor CHX and small interfering RNA of UPF1 (si-UPF1) significantly increased mRNA or protein expression of TSPAN12 in cells transfected with the mutant vectors.
CONCLUSIONS: Nine mutations in TSPAN12 gene are reported in 9 FEVR patients with a unique series of ocular abnormalities. The three novel TSPAN12 mutations trigger NMD would cause the decrease of TSPAN12 proteins that participate in biosynthesis and assembly of microfibers, which might lead to FEVR, and suggest that intronic sequence analysis might be a vital tool for genetic counseling and prenatal diagnoses.
摘要：
背景：报道新发现的具有家族性渗出性玻璃体视网膜病变（FEVR）独特形式的TSPAN12突变，并找出TSPAN12中重复的新型内含子变异导致FEVR的可能机制。
结果：通过基于面板的NGS检测到9个具有独特形式的FEVR的TSPAN12突变。MINI-Gene分析显示mRNA的两种剪接模式，处理两个不同的条带A和B，和突变型显示与Exon11跳跃的剪接模式的替换。野生型和突变型TSPAN12载体的构建显示过早终止密码子(PTC)的出现。体外表达检测显示，与野生型组相比，用突变载体转染的细胞中TSPAN12mRNA和蛋白质的表达水平显着下调。相反,翻译抑制剂CHX和UPF1的小干扰RNA（si-UPF1）显着增加了用突变载体转染的细胞中TSPAN12的mRNA或蛋白质表达。
结论：在9例FEVR患者中报道了TSPAN12基因的9个突变，这些患者具有一系列独特的眼部异常。三个新的TSPAN12突变触发NMD会导致参与微纤维生物合成和组装的TSPAN12蛋白的减少，这可能会导致FEVR，并表明内含子序列分析可能是遗传咨询和产前诊断的重要工具。
结果：通过基于面板的NGS检测到9个具有独特形式的FEVR的TSPAN12突变。MINI-Gene分析显示mRNA的两种剪接模式，处理两个不同的条带A和B，和突变型显示与Exon11跳跃的剪接模式的替换。野生型和突变型TSPAN12载体的构建显示过早终止密码子(PTC)的出现。体外表达检测显示，与野生型组相比，用突变载体转染的细胞中TSPAN12mRNA和蛋白质的表达水平显着下调。相反,翻译抑制剂CHX和UPF1的小干扰RNA（si-UPF1）显着增加了用突变载体转染的细胞中TSPAN12的mRNA或蛋白质表达。
结论：在9例FEVR患者中报道了TSPAN12基因的9个突变，这些患者具有一系列独特的眼部异常。三个新的TSPAN12突变触发NMD会导致参与微纤维生物合成和组装的TSPAN12蛋白的减少，这可能会导致FEVR，并表明内含子序列分析可能是遗传咨询和产前诊断的重要工具。
