Follow-up of two newborns with c.158G>A (p.Arg53His) mutation in<i><strong>PAH</strong></i> gene and assessment of the site function

doi:10.3724/zdxbyxb-2021-0256

J Zhejiang Univ (Med Sci)

2021, Vol. 50

Issue (4): 444-453 DOI: 10.3724/zdxbyxb-2021-0256

Follow-up of two newborns with c.158G>A (p.Arg53His) mutation inPAH gene and assessment of the site function

WANG Jie^1,²(

),ZHU Bo¹,ZHANG Lichun¹,ZHAO Yitong³,WANG Xiaohua^1,*,JIA Yueqi^1,*

1. Genetic Eugenic Department of Inner Mongolia Maternity and Child Health Care Hospital, Hohhot 010020, China;
2. State Key Laboratory of Reproductive Regulation & Breeding of Grass Land Livestock, College of Life Science, Inner Mongolia University, Hohhot 010021, China;
3. Medical Genetics and Prenatal Diagnosis of Sichuan Provincial Maternity and Child Health Care Hospital, Chengdu 610041, China

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Abstract

Objective: To investigate the clinical significance of PAHc.158G>A (p.Arg53His) mutation.Methods: The blood phenylalanine (Phe) was continuously monitored in 2 unrelated newborns with suspected hyperphenylalaninimia (HPA) carrying PAH c.158G>A mutation. The cross-species conservation of the mutant amino acid was analyzed using T-Coffee. Swiss-Model software was used to construct a 3D protein structure and the impact of candidate mutations on the secondary structure of the protein product was analyzed. The population carrying rate of the p.Arg53His mutation was analyzed by literature searching. Allelic phenotype values (APV) and genotypic phenotype values (GPV) were used to predict the phenotype associated with the mutation.Results:Two mutations of PAHgene were detected in each newborn: c.611A>G(p.Tyr204Cys), c.158G>A(p.Arg53His) and c.1238G>C(p.Arg413Pro), c.158G>A(p.Arg53His). Two children tolerated normal diet and plasma Phe levels were within the normal range during follow-up. The mother of case 2 was homozygous with p.Arg53His mutation under the condition of long-term normal diet, and the blood Phe concentration and Phe/Tyr were all within the normal range. The mutant amino acids were not highly conserved among the 13 different species. The 3D structural model showed that p.Arg53His mutation reduced the hydrogen bond from 2 to 1 between the 53rd and 49th amino acids of PAH. The allele frequency of p.Arg53His was 0.015?08 in HPA patients and 0.001?621 in normal population, while the prevalence of p.Arg53His allele was highest in the East Asian normal population (0.013?73). The APV and GPV system predicted that the mutation was related to mild HPA(MHP) type.Conclusion: The different compound heterozygous mutations of p.Arg53His lead to clinical phenotype varieties. The reduction of enzyme activity caused by the mutation of p.Arg53His is not sufficient to cause symptoms of phenylketonuria, so the mutation may be “likely benign”.

Key words： Hyperphenylalaninimia Phenylalanine hydroxylase deficiency p.Arg53His mutation Phenotype Phenylalanine Follow-up studies

Received: 03 May 2021 Published: 01 November 2021

CLC:	R722.1
	R596

Corresponding Authors: WANG Xiaohua,JIA Yueqi E-mail: wangjie8867@163.com

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Cite this article:

WANG Jie,ZHU Bo,ZHANG Lichun,ZHAO Yitong,WANG Xiaohua,JIA Yueqi. Follow-up of two newborns with c.158G>A (p.Arg53His) mutation inPAH gene and assessment of the site function. J Zhejiang Univ (Med Sci), 2021, 50(4): 444-453.

URL:

http://www.zjujournals.com/med/10.3724/zdxbyxb-2021-0256 OR http://www.zjujournals.com/med/Y2021/V50/I4/444

苯丙氨酸羟化酶基因c.158G>A(p.Arg53His)突变患儿随访及突变位点功能评估

目的：评价苯丙氨酸羟化酶（PAH）基因c.158G>A（p.Arg53His）突变的临床意义。方法：持续监测2例携带PAH基因p.Arg53His突变的疑似高苯丙氨酸血症患儿血液中苯丙氨酸（Phe）浓度，分析患儿的临床生化特征，应用T-Coffee系统分析PAH蛋白的跨种属保守性，应用Swiss-Model对正常结构及变异结构的PAH进行蛋白质三维结构建模及比对分析突变所致蛋白质空间结构的改变。检索现有数据库及文献统计p.Arg53His突变的人群携带率，应用等位基因表型值（APV）与基因型表型值（GPV）预测系统对该突变相关表型进行预测。结果：2例新生儿在PAH基因上分别检出两个突变：c.611A>G（p.Tyr204Cys）、c.158G>A（p.Arg53His）和c.1238G>C（p.Arg413Pro）、c.158G>A（p.Arg53His）。2例新生儿能耐受正常饮食，在随访期间血Phe水平在正常范围内。例2的母亲为p.Arg53His纯合突变，长期未进行低蛋白质、低Phe饮食干预，血Phe浓度、Phe/酪氨酸比值均在正常范围。突变的氨基酸在13个不同物种间并非高度保守。三维结构建模结果显示，p.Arg53His突变使得PAH第53位和第49位氨基酸之间的氢键由2个减少为1个，降低了二聚体的稳定性。p.Arg53His在高苯丙氨酸血症患者中的等位基因频率为0.015?08，在健康人群中的等位基因频率为0.001?621，其中东亚人群中的携带率最高，为0.013?73。APV与GPV系统预测结果显示，该突变与轻度高苯丙氨酸血症型别相关。结论：PAH基因p.Arg53His突变与不同突变组合为复合杂合状态可引起临床表型差异。p.Arg53His突变引起体内酶活性的降低不足以出现苯丙酮尿症临床症状，分类为“可能良性”。

关键词： 高苯丙氨酸血症, 苯丙氨酸羟化酶缺乏症, p.Arg53His突变, 表型, 苯丙氨酸, 随访研究

序号	性别	随访时间（月）	互补DNA 改变	氨基酸改变	初筛时Phe浓度（μmol/L）	Phe/Tyr比值	随访期间平均Phe浓度（ $x ˉ ± s$ ，μmol/L）	临床表现	干预措施
例1	男	10	c.[158G>A];[ 611A>G]	p.[R53H];[Y204C]	94.8	1.47	88±19	无	无
例2	男	12	c.[158G>A];[1238 G>C]	p.[R53H];[R413P]	122.4	1.70	110±20	无	无
例2母亲	女	—	c.[158G>A];[ 158G>A]	p.[R53H];[R53H]	69.6	1.56	—	无	无

Table 1 Characteristics of p.Arg53His (p.R53H) mutation carriers and their plasma phenylalanine levels during follow up

Figure 1 Cross-species conservation of the p.Arg53His mutation

Figure 2 3D structure prediction of PAH wild-type and p.Arg53His mutation

Table 2 Allele frequency of the p.Arg53His (p.R53H) mutation in patients with hyperphenylalaninimia

Table 3 Allele frequency of the p.Arg53His (p.R53H) mutation in the general population

Table 4 Biochemical phenotypes of patients with phenylalanine hydroxylase deficiency carrying the p.Arg53His (p.R53H) mutation (compand heterozygote)

Table 5 Biochemical phenotypes of patients with phenylalanine hydroxylase deficiency carrying the p.Arg53His (p.R53H) mutation (Zygosity of other mutation phase unknown)


[1]	BERCOVICHD, ELIMELECHA, YARDENIT, et al.A mutation analysis of the phenylalanine hydroxylase (PAH) gene in the Israeli population[J]Ann Hum Genet, 2008, 72( 3): 305-309. doi: 10.1111/j.1469-1809.2007.00425.x

[2]	中华医学会医学遗传学分会遗传病临床实践指南撰写组, 黄尚志, 宋昉. 苯丙酮尿症的临床实践指南[J]. 中华医学遗传学杂志, 2020, 37(3): 226-234 Writing Group for Practice Guidelines for Diagnosis and Treatment of Genetic Diseases, Medical Genetics Branch of Chinese Medical Association, HUANG Shangzhi, SONG Fang. Clinical practice guidelines for phenylketonuria[J]. Chinese Journal of Medical Genetics, 2020, 37(3): 226-234. (in Chinese)

[3]	REGIER D S, GREENE C L. Phenylalanine hydroxylase deficiency[J/OL]. (2017-01-05)[2021-04-12]. https://www.ncbi.nlm.nih.gov/books/NBK1504/

[4]	PARKY S, SEOUNGC S, LEES W, et al.Identification of three novel mutations in Korean phenylketonuria patients: R53H, N207D, and Y325X[J]Hum Mutat, 1998, 11( S1): S121-S122. doi: 10.1002/humu.1380110140

[5]	CHOIR, LEEJ, PARKH D, et al.Reassessing the significance of the PAH C.158g>a (P.Arg53His) variant in patients with hyperphenylalaninemia[J]J Pediatr Endocrinol Metab, 2017, 30( 11): 1211-1218. doi: 10.1515/jpem-2017-0158

[6]	OKANOY, KUDOS, NISHIY, et al.Molecular characterization of phenylketonuria and tetrahydrobiopterin-responsive phenylalanine hydroxylase deficiency in Japan[J]J Hum Genet, 2011, 56( 4): 306-312. doi: 10.1038/jhg.2011.10

[7]	TAOJ, LIN, JIAH, et al.Correlation between genotype and the tetrahydrobiopterin-responsive phenotype in Chinese patients with phenylketonuria[J]Pediatr Res, 2015, 78( 6): 691-699.. doi: 10.1038/pr.2015.167

[8]	ZHUT, YEJ, HANL, et al.Variations in genotype-phenotype correlations in phenylalanine hydroxylase deficiency in Chinese Han population[J]Gene, 2013, 529( 1): 80-87.. doi: 10.1016/j.gene.2013.07.079

[9]	POLAKE, FICEKA, RADVANSZKYJ, et al.Phenylalanine hydroxylase deficiency in the Slovak population: genotype-phenotype correlations and genotype-based predictions of BH4-responsiveness[J]Gene, 2013, 526( 2): 347-355. doi: 10.1016/j.gene.2013.05.057

[10]	BERCOVICHD, ELIMELECHA, ZLOTOGORAJ, et al.Genotype-phenotype correlations analysis of mutations in the phenylalanine hydroxylase (PAH) gene[J]J Hum Genet, 2008, 53( 5): 407-418. doi: 10.1007/s10038-008-0264-4

[11]	FIORIL, FIEGEB, RIVAE, et al.Incidence of Bh4-responsiveness in phenylalanine-hydroxylase-deficient Italian patients[J]Mol Genet Metab, 2005, 67-74. doi: 10.1016/j.ymgme.2005.06.017

[12]	SUY, WANGH, REJIAFUN, et al.The molecular epidemiology of hyperphenylalaninemia in uygur population: incidence from newborn screening and mutationalspectra[J]Ann Transl Med, 2019, 7( 12): 258. doi: 10.21037/atm.2019.05.16

[13]	LEED H, KOOS K, LEEK S, et al.The molecular basis of phenylketonuria in Koreans[J]J Hum Genet, 2004, 49( 11): 617-621. doi: 10.1007/s10038-004-0197-5

[14]	WETTSTEINS, UNDERHAUGJ, PEREZB, et al.Linking genotypes database with locus-specific database and genotype-phenotype correlation in phenylketonuria[J]Eur J Hum Genet, 2015, 23( 3): 302-309. doi: 10.1038/ejhg.2014.114

[15]	DATEKIS, WATANABES, NAKATOMIA, et al.Genetic background of hyperphenylalaninemia in Nagasaki, Japan[J]Pediatr Int, 2016, 58( 5): 431-433. doi: 10.1111/ped.12924

[16]	中华人民共和国卫生部. 新生儿疾病筛查技术规范(2010年版) [A/OL]. (2010-11-10)[2020-06-11]. http://www.nhc.gov.cn/cmsresources/mohfybjysqwss/cmsrsdocument/doc10798.doc Ministry of Health of the People’s Republic of China. Technical guide of newborn screening in China(2010)[A/OL]. (2010-11-10)[2020-06-11]. http://www.nhc.gov.cn/cmsresources/mohfybjysqwss/cmsrsdocument/doc10798.doc. (in Chinese)

[17]	RICHARDSS, AZIZN, BALES, 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): 405-423. doi: 10.1038/gim.2015.30

[18]	WANGR, SHENN, YEJ, et al.Mutation spectrum of hyperphenylalaninemia candidate genes and the genotype-phenotype correlation in the Chinese population[J]Clin Chim Acta, 2018, 132-138. doi: 10.1016/j.cca.2018.02.035

[19]	KIMS W, JUNGJ, OHH J, et al.Structural and functional analyses of mutations of the human phenylalanine hydroxylase gene[J]Clin Chim Acta, 2006, 365( 1-2): 279-287. doi: 10.1016/j.cca.2005.09.019

[20]	PEYA L, STRICHERF, SERRANOL, et al.Predicted effects of missense mutations on native-state stability account for phenotypic outcome in phenylketonuria, a paradigm of misfolding diseases[J]Am J Hum Genet, 2007, 81( 5): 1006-1024. doi: 10.1086/521879

[21]	GULDBERGP, REYF, ZSCHOCKEJ, et al.A european multicenter study of phenylalanine hydroxylase deficiency: classification of 105 mutations and a general system for genotype-based prediction of metabolic phenotype[J]Am J Hum Genet, 1998, 63( 1): 71-79. doi: 10.1086/301920

[22]	LIN, JIAH, LIUZ, et al.Molecular characterisation of phenylketonuria in a Chinese mainland population using next-generation sequencing[J]Sci Rep, 2015, 5( 1): 15769. doi: 10.1038/srep15769

[23]	SONGF, QUY J, ZHANGT, et al.Phenylketonuria mutations in northern China[J]Mol Genet Metab, 2005, 107-118. doi: 10.1016/j.ymgme.2005.09.001

[24]	ZHUT, QINS, YEJ, et al.Mutational spectrum of phenylketonuria in the Chinese Han population: a novel insight into the geographic distribution of the common mutations[J]Pediatr Res, 2010, 67( 3): 280-285. doi: 10.1203/PDR.0b013e3181c9fb85

[25]	LIANGY, HUANGM Z, CHENGC Y, et al.The mutation spectrum of the phenylalanine hydroxylase (PAH) gene and associated haplotypes reveal ethnic heterogeneity in the Taiwanese population[J]J Hum Genet, 2014, 59( 3): 145-152. doi: 10.1038/jhg.2013.136

[26]	JEANNESSON-THIVISOLE, FEILLETF, CHéRYC, et al.Genotype-phenotype associations in French patients with phenylketonuria and importance of genotype for full assessment of tetrahydrobiopterin responsiveness[J]Orphanet J Rare Dis, 2015, 10( 1): 158. doi: 10.1186/s13023-015-0375-x

[27]	AULEHLA-SCHOLZC, HEILBRONNERH. Mutational spectrum in german patients with phenylalanine hydroxylase deficiency[J]Hum Mutat, 2003, 21( 4): 399-400. doi: 10.1002/humu.9116

[28]	RAZIPOURM, ALAVINEJADE, SAJEDIS Z, et al.Genetic study of the PAH locus in the Iranian population: familial gene mutations and minihaplotypes[J]Metab Brain Dis, 2017, 32( 5): 1685-1691. doi: 10.1007/s11011-017-0048-7

[29]	杜玮, 杨桂芸, 陆薇冰, 等. 青岛市苯丙氨酸羟化酶缺乏症患儿基因突变分析[J]. 发育医学电子杂志, 2020, 8(1):30-34 DU Wei, YANG Guiyun, LU Weibing, et al. Study on mutations of gene in children with phenylalanine hydroxylase deficiency in Qingdao[J]. Journal of Developmental Medicine, 2020, 8(1): 30-34. (in Chinese)

[30]	金克勤, 胡苑. 742例育龄妇女PAH基因突变筛查结果分析[J]. 中国优生与遗传杂志, 2020, 28(7): 15-17 JIN Keqin, HU Yuan. Analysis of screening results of mutation of PAH gene in 742 fertile woman[J]. Chinese Journal of Birth Health & Heredity, 2020, 28(7): 15-17. (in Chinese)

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