Clinical, molecular and functional investigation on an infant with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD).
Clinical, molecular and functional investigation on an infant with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD).
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因 Citrin 缺乏引起的新生儿肝内胆汁淤积 (NICCD) 婴儿的临床、分子和功能研究
DOI：
10.1371/journal.pone.0089267
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Song YZ
中科院分区：
文献类型：
--
作者：
Zhang ZH;Lin WX;Deng M;Zhao ST;Zeng HS;Chen FP;Song YZ
背景与目的近10年来，SLC 25 A13基因分析为枸橼酸缺乏症（CD）的明确诊断提供了可靠依据。同时，这些研究也产生了一些尚待解决的问题，包括SLC 25 A13错义突变的致病性和突变c.615+5G>A的mRNA产物。本研究旨在探讨一种新的错义突变对Citrin蛋白的天冬氨酸/谷氨酸载体（AGC）功能的影响，并探讨在同一CD婴儿中c.615+5G>A的异常转录。方法与结果通过筛查SLC 25 A13突变和外显子测序，证实该患者为c.615+5G>A复合杂合子和新的c.1064G>A（p.Arg355Gln）突变。一个保留了整个内含子6的异常转录本，r。[615通过RT-PCR和cDNA测序，检测到一个由c.615 + 5G>A产生的突变体[615 + 1789 ins; 615 + 5g> a]（GenBank登录号KJ 128074）。在对新的错义突变c.1064G>A进行生物信息学分析后，对3株分别用重组载体和空载体转化的agc 1 Δ酵母菌进行了生长能力测定。除生物信息学致病性证据外，经突变重组体转化的agc 1 Δ菌株的生长能力与空载体转化的菌株相同，但在功能分析中显著低于正常对照。结论通过对SLC 25 A13基因的遗传学、转录和功能分析，明确诊断CD患儿。本研究为c.615+5G>A突变的剪接位点性质提供了直接的实验室证据，而新的c.1064G>A变异在生物信息学和功能上证明是一种致病性突变，丰富了SLC 25 A13突变谱。
Background and Objective SLC25A13 analysis has provided reliable evidences for the definitive diagnosis of citrin deficiency (CD) in the past decade. Meanwhile, these studies generated some issues yet to be resolved, including the pathogenicity of SLC25A13 missense mutations and the mRNA product from the mutation c.615+5G>A. This study aims to investigate the effect of a novel missense mutation on the aspartate/glutamate carrier (AGC) function of citrin protein, and to explore the aberrant transcript from c.615+5G>A in the same CD infant. Methods and Results By means of screening for prevalent SLC25A13 mutations and exons sequencing, the patient proved a compound heterozygote of c.615+5G>A and a novel c.1064G>A (p.Arg355Gln) mutation. An aberrant transcript with retention of the entire intron 6, r.[615+1_615+1789ins; 615+5 g>a] (GenBank accession number KJ128074), which was resulted from c.615+5G>A, was detected by RT-PCR and cDNA sequencing. After bioinformatic analyses of the novel missense mutation c.1064G>A, the growth abilities of three agc1Δ yeast strains were tested, which had been transformed with recombinant or empty vectors, respectively. Besides the bioinformatically pathogenic evidences, the growth ability of the agc1Δ strains transformed with mutant recombinant was the same as with empty vector, but significantly lower than that with normal control in functional analysis. Conclusions A CD infant was definitely diagnosed in this paper by a genetic, transcriptional and functional analysis of SLC25A13 gene. This study provided direct laboratory evidences supporting the splice-site nature of the c.615+5G>A mutation, and the novel c.1064G>A variation, which proved a pathogenic mutation bioinformatically and functionally, enriched the SLC25A13 mutation spectrum.
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影响因子：
8
作者：
Dimmock, David;Kobayashi, Keiko;Scaglia, Fernando
通讯作者：
Scaglia, Fernando
影响因子：
--
作者：
Jiang Guang-yu;Cheng Zhao-ming;Liu Kai-shan
通讯作者：
Liu Kai-shan
影响因子：
6.5
作者：
Lehner, F;Demmelmair, H;Koletzko, B
通讯作者：
Koletzko, B
影响因子：
2.6
作者：
GIETZ, RD;SCHIESTL, RH;WOODS, RA
通讯作者：
WOODS, RA
影响因子：
5.8
作者：
Green, Richard E.;Lewis, Benjamin P.;Brenner, Steven E.
通讯作者：
Brenner, Steven E.