Genetic analysis of newborns with abnormal metabolism of 3-hydroxyisovalerylcarnitine

doi:10.3785/j.issn.1008-9292.2019.08.07

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 Zhengyan^1,*(

)

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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Abstract

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 words： Metabolism, 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: 6506157@zju.edu.cn;zhaozy@zju.edu.cn

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	WU Dingwen
	LU Bin
	YANG Jianbin
	YANG Rulai
	HUANG Xinwen
	TONG Fan
	ZHENG Jing
	ZHAO Zhengyan

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.

URL:

http://www.zjujournals.com/med/10.3785/j.issn.1008-9292.2019.08.07 OR http://www.zjujournals.com/med/Y2019/V48/I4/390

3-羟基异戊酰基肉碱代谢异常新生儿遗传学分析

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MCCC1、MCCC2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: MCCC1、MCCC2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
^#以基因组突变频率数据库(gnomAD)为例.ACMG：美国医学遗传学与基因组学学会.
MCCC1	外显子1	c.57G > A	p.W19*	致病	无	无	无	1
MCCC1	内含子2	c.137-3C > A		意义未明	无	无	无	1
MCCC1	外显子3	c.227_228delTG	p.V76Gfs*3	致病	无	无	无	1
MCCC1	外显子4	c.313C > T	p.Q105*	致病	无	无	4.06×10^-6	1
MCCC1	外显子4	c.321C > A	p.Y107*	致病	无	无	无	1
MCCC1	外显子5	c.440T > G	p.I147S	意义未明	无	无	无	1
MCCC1	内含子6	c.639+2T > A		致病	致病	有	8.12×10^-6	10
MCCC1	外显子7	c.641G > T	p.G214V	意义未明	无	无	无	2
MCCC1	外显子8	c.820dupA	p.R274Kfs*4	致病	无	无	无	1
MCCC1	外显子8	c.863A > G	p.E288G	致病	意义未明	有	2.58×10^-5	2
MCCC1	内含子8	c.874-1G > A		致病	无	无	无	2
MCCC1	外显子9	c.881T > G	p.I294S	意义未明	无	无	无	1
MCCC1	内含子11	c.1267+2T > G		致病	无	无	无	1
MCCC1	外显子12	c.1304C > T	p.A435V	意义未明	无	有	4.06×10^-6	1
MCCC1	外显子12	c.1315G > A	p.V439M	意义未明	意义未明	有	8.12×10^-6	1
MCCC1	外显子12	c.1331G > A	p.R444H	可能致病	致病	有	2.84×10^-5	2
MCCC1	外显子14	c.1630delA	p.R544Dfs*2	致病	无	无	无	1
MCCC1	外显子14	c.1679dupA	p.N560Kfs*10	致病	致病	有	无	3
MCCC1	外显子15	c.1731G > C	p.Q577H	意义未明	无	无	无	1
MCCC1	外显子16	c.1759G > T	p.G587C	意义未明	无	无	4.07×10^-6	1
MCCC1	外显子16	c.1797delA	p.K599Nfs*13	致病	无	有	无	2
MCCC2	外显子2	c.130G > C	p.E44Q	意义未明	无	无	无	1
MCCC2	外显子2	c.157_159dupGTA	p.V53_N54insV	意义未明	无	无	无	1
MCCC2	内含子4	c.384-20A > G		意义未明	可能致病	无	4.06×10^-5	1
MCCC2	外显子6	c.562C > T	p.R188X*	可能致病	无	无	1.22×10^-5	2
MCCC2	内含子8	c.804-10T > G		意义未明	无	无	无	1
MCCC2	外显子11	c.1061C > T	p.T354I	意义未明	无	无	无	1

Tab 1 Mutations of MCCC1 and MCCC2

Fig 2 Distribution of C5-OH increased multiple in different neonatal groups

Tab 2 Cases of maternal 3-methylcrotonyl-coenzyme A carboxylase deficiency and abnormal 3-hydroxyisovalerylcarnitine levels during follow-up


[1]	KORMAN S H . Inborn errors of isoleucine degradation:a review[J]. Mol Genet Metab, 2006, 89 (4): 289- 299 doi: 10.1016/j.ymgme.2006.07.010

[2]	CATANZANO F , OMBRONE D , DI STEFANO C et al. The first case of mitochondrial acetoacetyl-CoA thiolase deficiency identified by expanded newborn metabolic screening in Italy:the importance of an integrated diagnostic approach[J]. J Inherit Metab Dis, 2010, 33 Suppl 3:S91- S94

[3]	RAMSAY J , MORTON J , NORRIS M et al. Organic acid disorders[J]. Ann Transl Med, 2018, 6 (24): 472 doi: 10.21037/atm.2018.12.39

[4]	KU C S , COOPER D N , POLYCHRONAKOS C et al. Exome sequencing:dual role as a discovery and diagnostic tool[J]. Ann Neurol, 2012, 71 (1): 5- 14 doi: 10.1002/ana.22647

[5]	RICHARDS S , AZIZ N , BALE S et al. Standards and guidelines for the interpretation of sequence variants:a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology[J]. Genet Med, 2015, 17 (5): 405- 424 doi: 10.1038/gim.2015.30

[6]	OMBRONE D , GIOCALIERE E , FORNI G et al. Expanded newborn screening by mass spectrometry:New tests, future perspectives[J]. Mass Spectrom Rev, 2016, 35 (1): 71- 84 doi: 10.1002/mas.21463

[7]	YUNUS Z M , RAHMAN S A , CHOYY S et al. Pilot study of newborn screening of inborn error of metabolism using tandem mass spectrometry in Malaysia:outcome and challenges[J]. J Pediatr Endocrinol Metab, 2016, 29 (9): 1031- 1039

[8]	黄新文, 杨建滨, 童凡 et al. 串联质谱技术对新生儿遗传代谢病的筛查及随访研究[J]. 中华儿科杂志, 2011, 49 (10): 765- 770 doi: 10.3760/cma.j.issn.0578-1310.2011.10.013

[9]	WOJCIK M H , WIERENGA K J , RODAN L H et al. Beta-ketothiolase deficiency presenting with metabolic stroke after a normal newborn screen in two individuals[J]. JIMD Rep, 2018, 39:45- 54

[10]	GRUNERT S C , SCHLATTER S M , SCHMITT R N et al. 3-Hydroxy-3-methylglutaryl-coenzyme A lyase deficiency:Clinical presentation and outcome in a series of 37 patients[J]. Mol Genet Metab, 2017, 121 (3): 206- 215 doi: 10.1016/j.ymgme.2017.05.014

[11]	BALASUBRAMANIAM S , LEWIS B , MOCK D M et al. Leigh-like syndrome due to homoplasmic m.8993T > G variant with hypocitrullinemia and unusual biochemical features suggestive of multiple carboxylase deficiency (MCD)[J]. JIMD Rep, 2017, 33:99- 107

[12]	YANG L , YANG J , ZHANG T et al. Identification of eight novel mutations and transcript analysis of two splicing mutations in Chinese newborns with MCC deficiency[J]. Clin Genet, 2015, 88 (5): 484- 488 doi: 10.1111/cge.12535

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