Molecular-genetic characterization of Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene
Mucopolysaccharidosis I (MPS I) is a rare hereditary autosomal-recessive metabolic disorder, which occurs due to the deficiency of the lysosomal enzyme α-L-iduronidase (IDUA; EC 3.2.1.76). There are three clinical forms of MPS I: Hurler syndrome, MPS I H; MIM # 607014, ORPHA 93473, Hurler/Scheie syn...
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Інститут молекулярної біології і генетики НАН України
2016
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nasplib_isofts_kiev_ua-123456789-1528502025-02-09T13:30:05Z Molecular-genetic characterization of Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene Молекулярно-генетична характеристика пацієнтів з мукополісахаридозом І типу з України: виявлення трьох нових мутацій в гені α-L-ідуронідази Молекулярно-генетическая характеристика пациентов с мукополисахаридозом I типа из Украины: виявление трёх новых мутаций в гене α-L-идуронидазы Trofimova, N.S. Olkhovich, N.V. Biomedicine Mucopolysaccharidosis I (MPS I) is a rare hereditary autosomal-recessive metabolic disorder, which occurs due to the deficiency of the lysosomal enzyme α-L-iduronidase (IDUA; EC 3.2.1.76). There are three clinical forms of MPS I: Hurler syndrome, MPS I H; MIM # 607014, ORPHA 93473, Hurler/Scheie syndrome, MPS I H/S; MIM # 607015, ORPHA 93476, Scheie syndrome, MPS I S; MIM # 607016, ORPHA 93474. Aim. To identify the spectrum of mutations in the IDUA gene in Ukrainian patients with MPS I. Methods. RFLP-analysis, automated sequencing. Results. We have identified 100 % (34/34) mutant alleles of the IDUA gene among 18 Ukrainian patients (one proband had a sibling with the identical genotype) with MPS I from 17 families. The spectrum of mutations in the IDUA gene in Ukrainian patients with MPS I is represented by six known missence mutations: p.Q70*, p.W402*, p.A75T, p.A327P, p.P533L, p.S633I; two deletions: c.1398delC and c.46_57del_12, one insertion: c.889_899_ins_12, and one mutation in the splicing zone IVS11ds+5G–A. Three new missence mutations were revealed by us in the IDUA gene: p.N372S, p.Q563P and p.S633*. Conclusions. Our results may be used for planning the most reasonable algorithm of the molecular-genetic analysis of Ukrainian patients with MPS I. Мукополісахаридоз І типу (МПС І) – рідкісне спадкове аутосомно-рецесивне метаболічне захворювання, яке виникає внаслідок дефекту лізосомального ферменту α-L-ідуронідази (IDUA; EC 3.2.1.76). Розрізняють 3 клінічних форми МПС І: синдром Гурлера, МПС І Г; MIM # 607014, ORPHA 93473, синдром Гурлер/Шейе, МПС І Г/Ш; MIM # 607015, ORPHA 93476, синдром Шейе, МПС І Ш; MIM # 607016, ORPHA 93474. Мета. Виявлення повного спектру мутацій в гені IDUA у пацієнтів з МПС І з України. Методи. ПДРФ-аналіз, метод прямого автоматичного секвенування по Сенгеру на аналізаторі ABI 3130 (Applied Biosystems). Результати. На підставі проведених нами досліджень було ідентифіковано 100 % (34/34) мутантних алелів гену IDUA серед 18 хворих (один пробанд мав сибса с ідентичним генотипом) на МПС І пацієнтів з 17 родин з України. Спектр мутацій в гені IDUA у пацієнтів з МПС І в Україні представлений 6 відомими місенс-мутаціями – p.Q70*, p.W402*, p.A75T, p.А327P, p.P533L, p.S633I; 2 делеціями – с.1398delC та c.46_57del_12, 1 інсерцією с.889_899_ins_12, та 1 мутацією в зоні сплайсінгу IVS11ds+5G–A. Нами було виявлено три нових місенс-мутації в гені IDUA: p.N372S, p.Q563P та p.S633*. Висновки. Дані проведеного молекулярно-генетичного аналізу мутацій в гені IDUA можуть бути використані при плануванні найбільш раціонального алгоритму молекулярно-генетичного аналізу пацієнтів з МПС І в Україні. Мукополисахаридоз І типа (МПС І) – редкое наследственное аутосомно-рецессивное метаболическое заболевание, которое возникает вследствие дефекта фермента α-L-идуронидазы (IDUA; EC 3.2.1.76). Разделяют 3 клинических формы МПС І: синдром Гурлера, МПС І Г; MIM # 607014, ORPHA 93473, синдром Гурлер/Шейе, МПС І Г/Ш; MIM # 607015, ORPHA 93476, синдром Шейе, МПС І Ш; MIM # 607016, ORPHA 93474. Цель. Выявление полного спектра мутаций в гене IDUA у пациентов с МПС I из Украины. Методы. ПДРФ-анализ, метод прямого автоматического секвенирования по Сенгеру на анализаторе ABI 3130 (Applied Biosystems). Результаты. На основании проведенных нами исследований было идентифицировано 100 % (34/34) мутантных аллелей гена IDUA среди 18 пациентов (два пробанда имели сибсов с идентичными генотипами) c МПС I из 17 семей из Украины. Спектр мутаций в гене IDUA у пациентов с МПС I в Украине представлен известными 6 миссенс-мутациями– p.Q70*, p.W402*, p.A75T, p.А327P, p.P533L, p.S633I, 2 делециями – с.1398delC и c.46_57del_12, 1 инсерцией – с.889_899_ins_12 и 1 мутацией в зоне сплайсинга IVS11ds+5G–A. Нами было найдено три новых миссенс-мутаций в гене IDUA: p.N372S, p.Q563P и p.S633*. Выводы. Данные проведенного молекулярно-генетического анализа мутаций в гене IDUA могут быть использованы при планировании наиболее рационального алгоритма молекулярно-генетического анализа пациентов с МПС I в Украине. 2016 Article Molecular-genetic characterization of Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene / N.S. Trofimova, N.V. Olkhovich // Вiopolymers and Cell. — 2016. — Т. 32, № 6. — С. 442-449. — Бібліогр.: 15 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.00093B https://nasplib.isofts.kiev.ua/handle/123456789/152850 616-056.7-07 en Вiopolymers and Cell application/pdf Інститут молекулярної біології і генетики НАН України |
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Biomedicine Biomedicine |
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Biomedicine Biomedicine Trofimova, N.S. Olkhovich, N.V. Molecular-genetic characterization of Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene Вiopolymers and Cell |
| description |
Mucopolysaccharidosis I (MPS I) is a rare hereditary autosomal-recessive metabolic disorder, which occurs due to the deficiency of the lysosomal enzyme α-L-iduronidase (IDUA; EC 3.2.1.76). There are three clinical forms of MPS I: Hurler syndrome, MPS I H; MIM # 607014, ORPHA 93473, Hurler/Scheie syndrome, MPS I H/S; MIM # 607015, ORPHA 93476, Scheie syndrome, MPS I S; MIM # 607016, ORPHA 93474. Aim. To identify the spectrum of mutations in the IDUA gene in Ukrainian patients with MPS I. Methods. RFLP-analysis, automated sequencing. Results. We have identified 100 % (34/34) mutant alleles of the IDUA gene among 18 Ukrainian patients (one proband had a sibling with the identical genotype) with MPS I from 17 families. The spectrum of mutations in the IDUA gene in Ukrainian patients with MPS I is represented by six known missence mutations: p.Q70*, p.W402*, p.A75T, p.A327P, p.P533L, p.S633I; two deletions: c.1398delC and c.46_57del_12, one insertion: c.889_899_ins_12, and one mutation in the splicing zone IVS11ds+5G–A. Three new missence mutations were revealed by us in the IDUA gene: p.N372S, p.Q563P and p.S633*. Conclusions. Our results may be used for planning the most reasonable algorithm of the molecular-genetic analysis of Ukrainian patients with MPS I. |
| format |
Article |
| author |
Trofimova, N.S. Olkhovich, N.V. |
| author_facet |
Trofimova, N.S. Olkhovich, N.V. |
| author_sort |
Trofimova, N.S. |
| title |
Molecular-genetic characterization of Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene |
| title_short |
Molecular-genetic characterization of Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene |
| title_full |
Molecular-genetic characterization of Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene |
| title_fullStr |
Molecular-genetic characterization of Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene |
| title_full_unstemmed |
Molecular-genetic characterization of Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene |
| title_sort |
molecular-genetic characterization of ukrainian patients with mucopolysaccharidosis i: identification of three new mutations in α-l-iduronidase gene |
| publisher |
Інститут молекулярної біології і генетики НАН України |
| publishDate |
2016 |
| topic_facet |
Biomedicine |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/152850 |
| citation_txt |
Molecular-genetic characterization of Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene / N.S. Trofimova, N.V. Olkhovich // Вiopolymers and Cell. — 2016. — Т. 32, № 6. — С. 442-449. — Бібліогр.: 15 назв. — англ. |
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Вiopolymers and Cell |
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442
N. S. Trofimova, N. V. Olkhovich
© 2016 N. S. Trofimova et al.; Published by the Institute of Molecular Biology and Genetics, NAS of Ukraine on behalf of Biopolymers and Cell.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/),
which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited
UDC 616-056.7-07
Molecular-genetic characterization of Ukrainian patients
with mucopolysaccharidosis I: identification of three new mutations
in α-L-iduronidase gene
N. S. Trofimova1,2, N. V. Olkhovich1,2
1 National Children’s Specialized Hospital Okhmatdyt, Ministry of Health of Ukraine
28/1, Chornovola Str., Kyiv, Ukraine, 01135
2 State Institute of Genetic and Regenerative Medicine, NAMS of Ukraine
67, Vyshhorodska Str., Kyiv, Ukraine, 04114
ntrofimofa@gmail.com
Mucopolysaccharidosis I (MPS I) is a rare hereditary autosomal-recessive metabolic disorder,which occurs
due to the deficiency of the lysosomal enzyme α-L-iduronidase (IDUA; EC 3.2.1.76). There are three clinical
forms of MPS I: Hurler syndrome, MPS I H; MIM # 607014, ORPHA 93473, Hurler/Scheie syndrome, MPS
I H/S; MIM # 607015, ORPHA 93476, Scheie syndrome, MPS I S; MIM # 607016, ORPHA 93474. Aim. To
identify the spectrum of mutations in the IDUA gene in Ukrainian patients with MPS I. Methods. RFLP-
analysis, automated sequencing. Results. We have identified 100 % (34/34) mutant alleles of the IDUA gene
among 18 Ukrainian patients (one proband had a sibling with the identical genotype) with MPS I from 17
families. The spectrum of mutations in the IDUA gene in Ukrainian patients with MPS I is represented by six
known missence mutations: p.Q70*, p.W402*, p.A75T, p.A327P, p.P533L, p.S633I; two deletions: c.1398delC
and c.46_57del_12, one insertion: c.889_899_ins_12, and one mutation in the splicing zone IVS11ds+5G–A.
Three new missence mutations were revealed by us in the IDUA gene: p.N372S, p.Q563P and p.
S633*. Conclusions. Our results may be used for planning the most reasonable algorithm of the molecular-
genetic analysis of Ukrainian patients with MPS I.
K e y w o r d s: mucopolysaccharidosis, Hurler syndrome, Scheie syndrome, α-L-iduronidase.
Introduction
Mucopolysaccharidosis I (MPS I) is a rare heredi-
tary autosomal-recessive metabolic disorder, which
occurs due to the deficiency of the lysosomal en-
zyme α-L-iduronidase (IDUA; EC 3.2.1.76), in-
volved in the multistage catabolism of dermatan sul-
phate (DS) and heparan sulphate (HS). The deficien-
cy in α-L-iduronidase leads to the accumulation of
non-degraded GAG substrate in the lysosomes of
cells of different organs and tissues [1, 2].
Although the clinical phenotype of MPS I is a con-
tinuous spectrum from severe forms to mild ones,
generally three clinical forms are distinguished: 1) se-
vere form (Hurler syndrome, MPS I H; MIM #
607014, ORPHA 93473), with the manifestation of
symptoms by the age of 12 months, life span of no
more than 10 years and mental retardation, which is
manifested at the age under three; 2) intermediate
form (Hurler-Scheie syndrome, MPS I H/S; MIM #
607015, ORPHA 93476), with the manifestation of
symptoms at the age from 1 to 6 years, longer life
span and no or mild mental retardation, which is never
manifested under 3; 3) mild form (Scheie syndrome,
MPS I S; MIM # 607016, ORPHA 93474), with the
manifestation of the first symptoms of the disease af-
Biomedicine ISSN 1993-6842 (on-line); ISSN 0233-7657 (print)
Biopolymers and Cell. 2016. Vol. 32. N 6. P 442–449
doi: http://dx.doi.org/10.7124/bc.00093B
mailto:ntrofimofa@gmail.com
443
Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene
ter the age of 5, unchanged life span and complete
absence of mental retardation [1].
The gene, encoding α-L-iduronidase (IDUA;
MIM # 252800), was mapped in locus 4p16.3; it
consists of 14 exons, separated by 13 introns. An
open reading frame of cDNA (ORF) is ~ 2 kilobases
long and encodes the polypeptide of 653 amino ac-
ids [3]. At present there are over 220 mutations,
identified and characterized in the IDUA gene, in-
cluding missence/nonsense mutations, small dele-
tions and insertions, four gross-deletions, one gross-
insertion, 36 splicing mutations and three complex
mutations (Human Gene Mutation Database-
HGMD, http://www.hgmd.cf.ac.uk/ac/index.php).
In general most mutations occur less than in 3 % of
cases or are single.
The missence mutations p.W402*, p.Q70*, p.P533R
and p.G51D are the most common for the gene IDUA.
Mutation p.W402* has about 50 % distribution fre-
quency in Northern Europe, Great Britain, North
America, and Spain, whereas its frequency in the
Russian Federation, Italy, and Brazil is 4 %, 11 %, and
20 % respectively, according to the estimates of differ-
ent researchers [4, 5]. On the contrary, mutation p.Q70*
occurs in Scandinavia and the Russian Federation
much more frequently (up to 50 %) than in other coun-
tries [6]. The missence mutation p.P533R of probable
North African origin is wide-spread in the Mediterranean
region, and has 13 % and 10 % of mutant alleles in the
IDUA gene in Italy and Spain, respectively. According
to the data of Italian researchers, the missence mutation
p.G51D is major for Italy only, and is registered with
the frequency of 13 % for mutant alleles in the IDUA
gene among Italian patients with mucopolysaccharido-
sis I [4]. The frequency of mutations in the IDUA gene
for Ukraine has not been defined yet.
The aim of our work was to identify the whole
spectrum of mutations in the IDUA gene in Ukrainian
patients with MPS I.
Materials and Methods
The material of the research was the blood of 17 pa-
tients with MPS I (7 boys and 10 girls), whose diag-
nosis was confirmed in the Clinical Genetics
Laboratory, NCSH OKHMATDYT of the Ministry of
Health of Ukraine, using biochemical methods with
the determination of the activity of α-L-iduronidase
enzyme in lysosomes, and the blood of their 37 rela-
tives (siblings and parents) from different regions of
Ukraine [7]. The age of the patients at the moment of
the diagnosis determination was 2 ± 1 years for Hurler
syndrome, 14 ± 1 years for Hurler-Scheie syndrome
and Scheie syndrome. One proband had a sibling with
the identical genotype, who was not taken into consi-
deration while identifying the mutation frequency.
For control of the pathogenicity of new mutations, the
blood samples of 100 volunteers aged 18 to 60 years
without any clinical signs of lysosomal pathology
were used. The parents of all the patients and volun-
teers gave their informed consent for the studies. The
bioethics committee approved the research.
DNAs were extracted from the whole blood using
the commercial NeoSorb kits (Neogen, Ukraine).
The identification of major mutations p.Q70* and
p.W402* was conducted using the RFLP-analysis as
described in our previous publication [7].
The identification of rare mutations in the IDUA
gene involved the method of direct automated se-
quencing by the Sanger’s method using ABI 3130
analyzer (Applied Biosystems) according to the
manufacturer’s protocol. The sequences of primers
for each of fourteen exons of the IDUA gene were
selected using Primer3 program (http://simgene.
com/Primer3). The analysis of sequencing results
was made using programs Sequencing Analysis,
v.1.1/3.1 (Applied Biosystems, Life Technologies
Corporation, USA), Chromas and Blast (http://www.
ncbi.nlm.nih.gov/blast). The analysis of pathogenic-
ity of new mutations was made using programs
PolyPhen2 and Provean (http://genetics.bwh.har-
vard.edu/pph2/, http://provean.jcvi.org/index.php).
Results and Discussion
The studies, conducted by us, allowed identifying
100 % (34/34) mutant alleles of the IDUA gene in
Ukrainian patients with MPS I (Table 1).
The most common mutation for the investigated
Ukrainian patients with MPS I was nonsense -muta-
http://www.hgmd.cf.ac.uk/ac/index.php
http://simgene.com/Primer3
http://simgene.com/Primer3
http://www.ncbi.nlm.nih.gov/blast
http://www.ncbi.nlm.nih.gov/blast
http://genetics.bwh.harvard.edu/pph2/
http://genetics.bwh.harvard.edu/pph2/
http://provean.jcvi.org/index.php
444
N. S. Trofimova, N. V. Olkhovich
tion p.Q70* – 11 alleles out of 34. The frequency of
this mutation is 32.4 % (11/34) which is close to the
indices of the distribution of this mutation in Europe
[6]. Two patients had this mutation in the homozy-
gous state, seven patients – in the heterozygous state.
The other major mutation p.W402* in the IDUA
gene was found in five patients with MPS I in the
heterozygous state, which is 14.7 % (5/34), i.e. its
distribution in Ukraine is close to the indices of
Italy [4]. Two compound heterozygotes p.Q70*/p.
W402* were revealed.
In addition to major mutations, the previously de-
scribed deletion с.1398delC was identified with high
frequency of 8.8% (3/34) in Ukraine, in homo- and
heterozygous state in the patients with MPS I.
Four previously revealed mutations were de-
scribed among the missence mutations, identified
during the analysis, in addition to major p.Q70* and
p.W402*: in exon 2 – p.A75T, in exon 8 – p.А327P,
in exon 11 – p.P533L, and in exon 14 – p.S633I. The
previously described mutation in the splicing zone
for exon 11 IVS11ds+5G–A was identified for one
patient in one allele. A previously known insertion of
12 basic pairs с.889_899_ins_12 was revealed in
exon 7 in four patients, two of them were siblings. A
previously described deletion of 12 bp in exon 1
c.46_57del_12 was found in one patient.
Additionally, three mutations in the IDUA gene,
which have been neither previously described nor in-
cluded into HGMD database, were identified by us,
namely, three missence mutations: p.N372S in exon 8
(in three alleles in two patients), p.Q563P in exon 12
(in one allele), and nonsense-mutation p.S633* in
exon 14 (in two alleles in one patient) (Fig. 1).
No changes in the sequence of nucleotides of the
IDUA gene were identified by us in the individuals
of the control group, i.e. they may be estimated as
mutations, and not as a polymorphic variant.
The detailed analysis of these mutations using
programs PolyPhen2 and Provean confirmed their
pathological character (Fig. 2).
A new missence mutation p.N372S, which was
identified in exon 8, leads to the replacement of as-
paragine for serine, due to which the length of pro-
tein does not change, but there is a disorder in the
functioning of the protein, which causes the change
in the splicing site. A new missence mutation p.
Q563P, which was identified in exon 12, results in
the replacement of glutamine for proline, which does
not have any significant impact on the protein func-
Table 1. The localization of the identified mutations in exons of the IDUA gene and their frequency
Nucleotide replacement
in the IDUA gene
Aminoacid replacement in the
molecule of α-L-iduronidase
Number
of exons
Number of mutant alleles
in the ratio to the general
number of alleles
Reference to the first description of
the mutation in the IDUA gene
c.46_57del_12 p.S16_A19del 1 1/34 Bunge et al., 1994, [8]
c.208C>T p.Q70* 2 11/34 Clarke et al., 1993, [9]
c.223G>A p.A75T 2 1/34 Clarke et al., 1994, [10]
с.889_899_ins_12 p.T296_T299dup 7 3/34 Bunge et al., 1995, [11]
c.979G>C p.А327P 8 1/34 Bunge et al., 1995, [11]
c.1115A>G p.N372S 8 3/34 Our studies
c.1205G>A p.W402* 9 5/34 Scott et al., 1992, [12]
с.1398delС p.P467Rfs*58 9 3/34 Bertola et al., 2011, [4]
c.1598C>T p.P533L 11 1/34 Voskoboeva et al., 1998, [13]
IVS11ds+5G–A 1738+5G>A Int.11 1/34 Verturi et al., 2002, [14]
с.1688A>C p.Q563P 12 1/34 Our studies
c.1898C>T p.S633I 14 1/34 Beesley et al., 2001, [15]
с.1898C>A p.S633* 14 2/34 Our studies
445
Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene
tion, leading only to the point replacement of the
aminoacid in its structure. A new nonsense-mutation
p.S633* in exon 14 forms a stop-codon and shortens
the protein by 21 aminoacids (~10 % from the total
length of the protein).
Therefore, the spectrum of mutations in the IDUA
gene in Ukrainian patients with MPS I at presen is
represented by 9 missence mutations, 2 deletions of
1 and 12 bp, 1 insertion of 12 bp, and 1 mutation in
the splicing zone (Fig. 3).
Similar to many other European populations, two
mutations may be considered to be major for
Ukrainian patients with MPS I: p.Q70* (32.35 %)
and p.W402* (14.7 %). As for rare mutations, the
most common ones were found to be deletion
с.1398delC (8.8 %) and a new missence mutation
p.N372S (8.8 %), which were revealed in the homo-
A B C
D
Fig. 1. The chromatograms of the DNA sequence of new mutations in the
IDUA gene:
А – missence mutation p.N372S in the homozygous state in exon 8;
B – missence mutation p.N372S in the heterozygous state in exon 8;
C – missence mutation p.Q563P in the heterozygous state in exon 12;
D – missence mutation p.S633* in the homozygous state in exon 14.
A
B
Fig. 2. The conservatism of new missence mutations in our
studies:
А – new mutation p.Q563P (с.1688A>C);
B – new mutation p.N372S (c.1115A>G).
446
N. S. Trofimova, N. V. Olkhovich
and heterozygous state among Ukrainian patients
with MPS I. The insertion с.889_899_ins_12
(8.8 %), identified in four patients in the heterozy-
gous state, two of whom were siblings, was also
found to be quite common. A new nonsense muta-
tion p.S633* (5.9 %), which was identified in one
patient, may also potentially be more common for
Ukraine, as it was identified in the homozygous
state. However, this may also be a result of the intra-
familial accumulation of the pathological mutation
after the marriage of close relatives, who are parents
of this patient. Other mutations were represented by
single cases. Probably, these frequencies may change
with further identification of new patients. The most
rare mutations, identified by us in Ukrainian patients
with MPS I, were localized in exons 2, 7, 8, 9, and
14, and one in each – in exons 1, 11, and 12 of the
IDUA gene.
The complete characterization of the patients with
MPS I with the consideration of the clinical form of
the disease is presented in Table 2.
The results of molecular-genetic analysis of
Ukrainian patients with MPS I, obtained by us, dem-
onstrated that all the patients with major mutation
p.Q70* in the homozygous state, and two patients,
who were found to be compound heterozygotes of
p.Q70X*/ p.W402*, have a severe form of the dis-
ease – Hurler syndrome, which is in good agreement
with the data of other researchers about the associa-
tion of major nonsense -mutations p.Q70* and
p.W402* with the severe disease progression [6, 8].
The severe progression of MPS I was also noted for
patients with the major mutation p.W402* in one al-
lele and the missence mutation p.A75T (1 person)
[15]. A previously known single-nucleotide deletion
с.1398delC, identified in two patients in both the ho-
mozygous state and the compound with the major
mutation p.W402*, also characterized the presence
of the severe clinical form of MPS I, Hurler syn-
drome, in patients [4].
Two patients, who had a new, previously not de-
scribed missence mutation p.N372S in the homozygous
state and in the compound with the major mutation p.
Q70*, were characterized by early onset of the disease
and a severe phenotype. Three patients with the com-
pounds combination of major mutations and rare muta-
tions (genotypes p.Q70*/p.P533L, p.Q70*/c.46_57_
del_12 and p.W402*/p.S633I) had a severe form of the
disease – Hurler syndrome with early onset of the di-
sease.
Hurler-Scheie syndrome, which was identified in
two patients, was associated with genotypes
IVS11ds+5G-A/с.889_899_ins_12 and a new muta-
tion in the homozygous state p.S633*/p.S633*.
Scheie syndrome was diagnosed in four pa-
tients, two of whom were siblings, with geno-
types с.889_899_ins_12/p.Q70* – in three pa-
tients, and the missence mutation p.А327P in the
Fig. 2. The conservatism of new missence mutations in our studies:
А – new mutation p.Q563P (с.1688A>C);
B – new mutation p.N372S (c.1115A>G).
Fig. 3. The localization of mu-
tations in separate exons of the
IDUA gene in Ukrainian pa-
tients with MPS I
447
Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene
compound with the new mutation p.Q563P – in
one patient.
Therefore, our analysis of the molecular-genetic
characterization of Ukrainian patients with MPS I
allowed identifying 100 % mutant alleles. High fre-
quency of mutations p.Q70* (32.35 %) and p.W402*
(14.7 %) among Ukrainian patients substantiates
RFLP-screening for the presence of these mutations
at the first stage of the molecular-genetic analysis. It
would be reasonable to start the analysis during the
identification of rare mutations, which have led to
the development of the disease in the proband, with
“hot” regions of the IDUA gene – exons 2, 7, 8, 9,
and 14, where most pathogenic mutations were lo-
calized in the patients, studied by us. During the
planning of the analysis, this order would allow op-
timizing time and resources for the analysis and en-
suring the maximal efficiency of the molecular ge-
netic diagnostics of MPS I in Ukraine.
Conclusions
The molecular genetic analysis of pathogenic muta-
tions in the IDUA gene in 18 Ukrainian patients with
MPS I from 17 families (34 alleles) revealed 100 %
mutant alleles.
The share of the alleles with the major mutation
p.Q70* was the largest – 32.35 % (11/34). The other
major mutation for European populations, p.W402*,
was found in five patients (14.7 %) in one allele, so
it is also major for Ukraine.
Most rare mutations, identified by us in Ukrainian
patients with MPS I, were localized in exons 2, 7, 8,
9, and 14 of the IDUA gene, and one in each – in
exons 1, 11, and 12.
Three previously not described missence muta-
tions in the IDUA gene – p.N372S, p.Q563P and
p.S633* – were also revealed.
The data of the conducted molecular genetic anal-
ysis of mutations in the IDUA gene may be used
Table 2. The characterization of patients with MPS I
# Gender Age at the moment of diagnosis
determination Phenotype First allele Second allele
1 F 1.5 y.o. H p.Q70* p.Q70*
2 M 3.5 y.o. H p.Q70* p.Q70*
3 F 1 y.o. H p.Q70* p.P533L
4 M 2.5 y.o. H p.Q70* c.46_57_del_12
5 F 3 y.o. H p.Q70* p.N372S
6 M 2.5 y.o. H p.Q70* p.W402*
7 F 3 y.o. H p.Q70* p.W402*
8 M 2 y.o. H p.S633I p.W402*
9 F 2 y.o. H p.A75T p.W402*
10 F 4.5 y.o. H c.1398delC p.W402*
11 F 2 y.o. H c.1398delC c.1398delC
12 F 4 y.o. H p.N372S p.N372S
13 F 14 y.o. H/S p.S633* p.S633*
14 M 15 y.o. H/S IVS11ds+5G-A с.889_899_ins_12
15 M 9 y.o. S p.Q70* с.889_899_ins_12
16 M 14 y.o. S p.Q70* с.889_899_ins_12
17 F 8 y.o. S p.А327P p.Q563P
H – Hurler syndrome, H/S – Hurler-Scheie syndrome, S – Scheie syndrome;
F – girls, M – boys;
Mutations highlighted in bold are new mutations, which have not been described in the literature.
448
N. S. Trofimova, N. V. Olkhovich
while planning the most reasonable algorithm of the
molecular genetic analysis of Ukrainian patients
with MPS I in order to enhance the quality of provid-
ing medical care to the families, suffering from this
disease.
Acknowledgments
We are grateful to the clinical doctors of the Center
of Orphan Diseases and the Medical Genetics Center
of the National Children’s Specialized Hospital
Okhmatdyt for their providing clinical profiles of pa-
tients.
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Молекулярно-генетична характеристика пацієнтів з
мукополісахаридозом І типу з України: виявлення
трьох нових мутацій в гені α-L-ідуронідази
Н. С. Трофімова, Н. В. Ольхович
Мукополісахаридоз І типу (МПС І) – рідкісне спадкове ауто-
сомно-рецесивне метаболічне захворювання, яке виникає вна-
слідок дефекту лізосомального ферменту α-L-ідуронідази
(IDUA; EC 3.2.1.76). Розрізняють 3 клінічних форми МПС І:
синдром Гурлера, МПС І Г; MIM # 607014, ORPHA 93473, син-
дром Гурлер/Шейе, МПС І Г/Ш; MIM # 607015, ORPHA 93476,
синдром Шейе, МПС І Ш; MIM # 607016, ORPHA 93474.
Мета. Виявлення повного спектру мутацій в гені IDUA у паці-
єнтів з МПС І з України. Методи. ПДРФ-аналіз, метод прямого
автоматичного секвенування по Сенгеру на аналізаторі ABI
3130 (Applied Biosystems). Результати. На підставі проведе-
них нами досліджень було ідентифіковано 100% (34/34) му-
449
Ukrainian patients with mucopolysaccharidosis I: identification of three new mutations in α-L-iduronidase gene
тантних алелів гену IDUA серед 18 хворих (один пробанд мав
сибса с ідентичним генотипом) на МПС І пацієнтів з 17 родин
з України. Спектр мутацій в гені IDUA у пацієнтів з МПС І в
Україні представлений 6 відомими місенс-мутаціями – p.Q70*,
p.W402*, p.A75T, p.А327P, p.P533L, p.S633I; 2 делеціями –
с.1398delC та c.46_57del_12, 1 інсерцією с.889_899_ins_12, та
1 мутацією в зоні сплайсінгу IVS11ds+5G–A. Нами було вияв-
лено три нових місенс-мутації в гені IDUA: p.N372S, p.Q563P
та p.S633*. Висновки. Дані проведеного молекулярно-гене-
тичного аналізу мутацій в гені IDUA можуть бути використані
при плануванні найбільш раціонального алгоритму молеку-
лярно-генетичного аналізу пацієнтів з МПС І в Україні.
К л юч ов і с л ов а: мукополісахаридоз, синдром Гурлер,
синдром Шейе, α-L-ідуронідаза.
Молекулярно-генетическая характеристика пациентов
с мукополисахаридозом I типа из Украины: виявление
трёх новых мутаций в гене α-L-идуронидазы
Н. С. Трофимова, Н. В. Ольхович
Мукополисахаридоз І типа (МПС І) – редкое наследственное
аутосомно-рецессивное метаболическое заболевание, которое
возникает вследствие дефекта фермента α-L-идуронидазы
(IDUA; EC 3.2.1.76). Разделяют 3 клинических формы МПС І:
синдром Гурлера, МПС І Г; MIM # 607014, ORPHA 93473, син-
дром Гурлер/Шейе, МПС І Г/Ш; MIM # 607015, ORPHA 93476,
синдром Шейе, МПС І Ш; MIM # 607016, ORPHA 93474.
Цель. Выявление полного спектра мутаций в гене IDUA у па-
циентов с МПС I из Украины. Методы. ПДРФ-анализ, метод
прямого автоматического секвенирования по Сенгеру на ана-
лизаторе ABI 3130 (Applied Biosystems). Результаты. На осно-
вании проведенных нами исследований было идентифициро-
вано 100% (34/34) мутантных аллелей гена IDUA среди 18 па-
циентов (два пробанда имели сибсов с идентичными генотипа-
ми) c МПС I из 17 семей из Украины. Спектр мутаций в гене
IDUA у пациентов с МПС I в Украине представлен известными
6 миссенс-мутациями - p.Q70*, p.W402*, p.A75T, p.А327P, p.
P533L, p.S633I, 2 делециями – с.1398delC и c.46_57del_12,
1 инсерцией – с.889_899_ins_12 и 1 мутацией в зоне сплайсин-
га IVS11ds+5G–A. Нами было найдено три новых миссенс-му-
таций в гене IDUA: p.N372S, p.Q563P и p.S633*. Выводы.
Данные проведенного молекулярно-генетического анализа му-
таций в гене IDUA могут быть использованы при планирова-
нии наиболее рационального алгоритма молекулярно-генети-
ческого анализа пациентов с МПС I в Украине.
К л юч е в ы е с л ов а: мукополисахаридоз, синдром Гурлер,
синдром Шейе, α-L-идуронидаза.
Received 10.10.2016
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