Department of Pediatrics, Juntendo University Graduate School of Medicine, Tokyo, Japan.
Department of Biochemistry, Juntendo University Graduate School of Medicine, Tokyo, Japan.
PLoS One. 2020 Nov 9;15(11):e0241869. doi: 10.1371/journal.pone.0241869. eCollection 2020.
Chronic enteropathy associated with SLCO2A1 gene (CEAS) is caused by loss-of-function mutations in SLCO2A1, which encodes a prostaglandin (PG) transporter. In this study, we report a sibling case of CEAS with a novel pathogenic variant of the SLCO2A1 gene. Compound heterozygous variants in SLCO2A1 were identified in an 8-year-old boy and 12-year-old girl, and multiple chronic nonspecific ulcers were observed in the patients using capsule endoscopy. The splice site mutation (c.940 + 1G>A) of the paternal allele was previously reported to be pathogenic, whereas the missense variant (c.1688T>C) of the maternal allele was novel and had not yet been reported. The affected residue (p.Leu563Pro) is located in the 11th transmembrane domain (helix 11) of SLCO2A1. Because SLCO2A1 mediates the uptake and clearance of PGs, the urinary PG metabolites were measured by liquid chromatography coupled to tandem mass spectrometry. The urinary tetranor-prostaglandin E metabolite levels in the patients were significantly higher than those in unaffected individuals. We established cell lines with doxycycline-inducible expression of wild type SLCO2A1 (WT-SLCO2A1) and the L563P mutant. Immunofluorescence staining showed that WT-SLCO2A1 and the L563P mutant were dominantly expressed on the plasma membranes of these cells. Cells expressing WT-SLCO2A1 exhibited time- and dose-dependent uptake of PGE2, while the mutant did not show any uptake activity. Residue L563 is very close to the putative substrate-binding site in SLCO2A1, R561 in helix 11. However, in a molecular model of SLCO2A1, the side chain of L563 projected outside of helix 11, indicating that L563 is likely not directly involved in substrate binding. Instead, the substitution of Pro may twist the helix and impair the transporter function. In summary, we identified a novel pathogenic variant of SLCO2A1 that caused loss-of-function and induced CEAS.
与 SLCO2A1 基因相关的慢性肠病(CEAS)是由 SLCO2A1 基因功能丧失突变引起的,该基因编码一种前列腺素(PG)转运体。在本研究中,我们报告了一例由 SLCO2A1 基因新的致病性变异引起的 CEAS 同胞病例。在一名 8 岁男孩和一名 12 岁女孩中发现了 SLCO2A1 的复合杂合变异,患者使用胶囊内镜观察到多个慢性非特异性溃疡。父系等位基因的剪接位点突变(c.940 + 1G>A)先前被报道为致病性的,而母系等位基因的错义变异(c.1688T>C)是新的,尚未报道。受影响的残基(p.Leu563Pro)位于 SLCO2A1 的第 11 个跨膜域(螺旋 11)中。由于 SLCO2A1 介导 PG 的摄取和清除,通过液相色谱-串联质谱法测量尿 PG 代谢物。患者的尿四氢前列腺素 E 代谢物水平明显高于未受影响的个体。我们建立了具有诱导表达野生型 SLCO2A1(WT-SLCO2A1)和 L563P 突变体的细胞系。免疫荧光染色显示,WT-SLCO2A1 和 L563P 突变体在这些细胞的质膜上呈优势表达。表达 WT-SLCO2A1 的细胞表现出时间和剂量依赖性的 PGE2 摄取,而突变体则没有表现出任何摄取活性。残基 L563 非常接近 SLCO2A1 中假定的底物结合位点,即螺旋 11 中的 R561。然而,在 SLCO2A1 的分子模型中,L563 的侧链突出于螺旋 11 之外,表明 L563 不太可能直接参与底物结合。相反,脯氨酸的取代可能会扭曲螺旋并损害转运体功能。总之,我们鉴定了一种新的 SLCO2A1 致病性变异,导致功能丧失并引起 CEAS。