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J Zhejiang Univ (Med Sci)  2019, Vol. 48 Issue (4): 390-396    DOI: 10.3785/j.issn.1008-9292.2019.08.07
Genetic analysis of newborns with abnormal metabolism of 3-hydroxyisovalerylcarnitine
WU Dingwen1(),LU Bin2,YANG Jianbin1,YANG Rulai1,HUANG Xinwen1,TONG Fan1,ZHENG Jing1,ZHAO Zhengyan1,*()
1. Zhejiang Neonatal Screening Center, Department of Genetics and Metabolism, the Children's Hospital, Zhejiang University School of Medicine, Hangzhou 310052, China
2. Zhejiang Biosan Biochemical Technologies Co. Ltd, Hangzhou 310012, China
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Objective: To investigate the genetic characterization of 3-hydroxyisovalerylcarnitine (C5-OH) metabolic abnormality in neonates. Methods: Fifty two newborns with increased C5-OH, C5-OH/C3 and C5-OH/C8 detected by tandem mass spectrometry during neonatal screening were enrolled in the study. Genomic DNA was extracted from the whole blood samples of 52 cases and their parents. Seventy-nine genes associated with genetic and metabolic diseases including MCCC1, MCCC2 were targeted by liquid capture technique. Variation information of these genes was examined by high-throughput sequencing and bioinformatic analysis, and then was classified based on the American College of Medical Genetics and Genomics (ACMG) standards and guidelines. The genetic types were classified as wild-type, MCCC1-maternal-mutation, MCCC1-paternal-mutation and MCCC2-mutation. Wilcoxon rank-sum test was performed for the increased multiples of C5-OH calculated in neonatal screening. Results: Twenty one MCCC1 variants (14 novel) were identified in 37 cases, 6 MCCC2 variants (5 novel) in 4 cases. The increased multiple of C5-OH calculated in MCCC1-maternal-mutation and MCCC2-mutation groups were significantly higher than that in wild-type group (all P < 0.05), while there was no significant difference between MCCC1-paternal-mutation group and wild-type group (P>0.05). Conclusion: Mutations on MCCC1 and MCCC2 genes are the major genetic causes for the increased C5-OH in neonates, and maternal single heterozygous mutation can contribute to the moderately to severely increased C5-OH.

Key wordsMetabolism, inborn errors/blood      Organic acids/blood      Acyl coenzyme A/deficiency      Genes/genetics      Mass spectrometry      Neonatal screening     
Received: 05 March 2019      Published: 30 October 2019
CLC:  R722.11  
Corresponding Authors: ZHAO Zhengyan     E-mail:;
Cite this article:

WU Dingwen,LU Bin,YANG Jianbin,YANG Rulai,HUANG Xinwen,TONG Fan,ZHENG Jing,ZHAO Zhengyan. Genetic analysis of newborns with abnormal metabolism of 3-hydroxyisovalerylcarnitine. J Zhejiang Univ (Med Sci), 2019, 48(4): 390-396.

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目的: 探讨新生儿3-羟基异戊酰基肉碱(C5-OH)代谢异常的遗传学原因。方法: 收集2018年1月至12月在浙江省新生儿遗传代谢病筛查中心经串联质谱法筛查结果为C5-OH增高的52例新生儿的资料,包括新生儿筛查与复查随访的C5-OH、C5-OH/C3、C5-OH/C8检测数据,并换算成C5-OH增高倍数。采用液相捕获技术靶向捕获MCCC1MCCC2等79个遗传代谢病相关基因,通过高通量测序和生物信息学分析获取基因的突变信息,参考美国医学遗传学与基因组学学会(ACMG)分类标准进行分级。依据基因检测情况,将C5-OH增高新生儿分为未检出突变组、MCCC1母源突变组、MCCC1父源突变组、MCCC2突变组,采用威尔科克森秩和检验分析不同组间C5-OH增高倍数的差异。结果: 37例检出MCCC1突变,涉及21种突变型,其中14种为新发现的突变型;4例检出MCCC2突变,涉及6种突变型,其中5种为新发现的突变型。MCCC1母源突变组、MCCC2突变组的C5-OH增高倍数均高于未检出突变组(均P < 0.05),MCCC1父源突变组的C5-OH增高倍数与未检出突变组差异无统计学意义(P>0.05)。结论: MCCC1MCCC2基因突变是导致新生儿血C5-OH增高的主要遗传学原因,其中母源性单杂合突变可导致中重度C5-OH增高。

关键词: 代谢缺陷, 先天性/血液,  有机酸类/血液,  酰基辅酶A/缺乏,  基因/遗传学,  质谱分析法,  新生儿筛查 
Fig 1 Distribution of MCCC1 mutation sites
基因 区域 碱基 氨基酸 ACMG分类 数据库 文献 人群频率# 检出次数
MCCC1 外显子1 c.57G > A p.W19* 致病 1
MCCC1 内含子2 c.137-3C > A 意义未明 1
MCCC1 外显子3 c.227_228delTG p.V76Gfs*3 致病 1
MCCC1 外显子4 c.313C > T p.Q105* 致病 4.06×10-6 1
MCCC1 外显子4 c.321C > A p.Y107* 致病 1
MCCC1 外显子5 c.440T > G p.I147S 意义未明 1
MCCC1 内含子6 c.639+2T > A 致病 致病 8.12×10-6 10
MCCC1 外显子7 c.641G > T p.G214V 意义未明 2
MCCC1 外显子8 c.820dupA p.R274Kfs*4 致病 1
MCCC1 外显子8 c.863A > G p.E288G 致病 意义未明 2.58×10-5 2
MCCC1 内含子8 c.874-1G > A 致病 2
MCCC1 外显子9 c.881T > G p.I294S 意义未明 1
MCCC1 内含子11 c.1267+2T > G 致病 1
MCCC1 外显子12 c.1304C > T p.A435V 意义未明 4.06×10-6 1
MCCC1 外显子12 c.1315G > A p.V439M 意义未明 意义未明 8.12×10-6 1
MCCC1 外显子12 c.1331G > A p.R444H 可能致病 致病 2.84×10-5 2
MCCC1 外显子14 c.1630delA p.R544Dfs*2 致病 1
MCCC1 外显子14 c.1679dupA p.N560Kfs*10 致病 致病 3
MCCC1 外显子15 c.1731G > C p.Q577H 意义未明 1
MCCC1 外显子16 c.1759G > T p.G587C 意义未明 4.07×10-6 1
MCCC1 外显子16 c.1797delA p.K599Nfs*13 致病 2
MCCC2 外显子2 c.130G > C p.E44Q 意义未明 1
MCCC2 外显子2 c.157_159dupGTA p.V53_N54insV 意义未明 1
MCCC2 内含子4 c.384-20A > G 意义未明 可能致病 4.06×10-5 1
MCCC2 外显子6 c.562C > T p.R188X* 可能致病 1.22×10-5 2
MCCC2 内含子8 c.804-10T > G 意义未明 1
MCCC2 外显子11 c.1061C > T p.T354I 意义未明 1
Tab 1 Mutations of MCCC1 and MCCC2
Fig 2 Distribution of C5-OH increased multiple in different neonatal groups
序号 C5-OH增高倍数 随访时间(月) 转归趋势 基因 突变位点 遗传来源 临床意见
初筛 随访 母亲
MCCC:甲基巴豆酰辅酶A羧化酶;3MCCD:3-甲基巴豆酰辅酶A羧化酶缺乏症; C5-OH:3-羟基异戊酰基肉碱.
1 24.14 1.01~21.60 169.64 10 逐步下降 MCCC1 c.440T > G 母亲 母源性3MCCD
2 13.98 0.92~7.53 119.13 6 逐步下降 MCCC1 c.639+2T > A 母亲 母源性3MCCD
3 15.19 1.25~10.66 166.43 6 逐步下降 MCCC1 c.639+2T > A 母亲 母源性3MCCD
4 18.20 2.15~19.92 189.20 2 逐步下降 MCCC1 c.1759G > T 母亲 母源性3MCCD
5 19.42 1.76~17.17 154.26 2 逐步下降 MCCC1 c.137-3C > A 母亲 母源性3MCCD
6 16.98 38.54~74.82 1.00 2 逐步下降 MCCC1 c.863A > G 母亲 原因不明,继续随访
7 19.65 63.61~99.87 2.65 11 持续增高 MCCC2 c.562C > T、c.1061C > T 母亲与父亲 原发性3MCCD
8 1.67 2.10~5.69 1.92 10 波动 MCCC2 c.384-20A > G、c.804-10T > G 母亲与父亲 原因不明,继续随访
9 5.41 4.5~5.85 1.66 2 维持 MCCC2 c.130G > C、c.157_159dupGTA 父亲与母亲 原因不明,继续随访
Tab 2 Cases of maternal 3-methylcrotonyl-coenzyme A carboxylase deficiency and abnormal 3-hydroxyisovalerylcarnitine levels during follow-up
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