Journal of Genetics, Vol. 96, No. 4, September 2017, pp.

583–589 © Indian Academy of Sciences

DOI 10.1007/s12041-017-0821-8

RESEARCH ARTICLE

GBA mutations in Gaucher type I Venezuelan patients: ethnic origins

and frequencies

GILBERTO GÓMEZ1 , SERGIO ARIAS1 , LEONOR CÁRDENAS2 , DALAL ZOGHBI2 and IRENE PARADISI1∗

1 Laboratory of Human Genetics, Venezuelan Institute for Scientific Research (IVIC), Caracas 10200-A, Venezuela
2 Hematology Service, Miguel Pérez Carreño Hospital, Caracas 10200-A, Venezuela
*For correspondence. E-mail: [email protected], [email protected].

Received 21 June 2016; revised 23 September 2016; accepted 5 December 2016; published online 8 September 2017

Abstract. Gaucher disease (GD), the most frequent lysosomal storage disease, is caused by heterogeneous mutations in the locus
coding for glucocerebrosidase (GBA). It is an autosomal recessive disorder with different phenotypes of which the most frequent is
the nonneuronopathic or type 1, prevalent worldwide. To date, more than 430 mutations have been described, but their frequency
distribution varies in different populations with four, N370S, L444P, IVS2 + 1G > A and 84insG, being the most frequent ones.
In Venezuela, 20 unrelated index cases with GD type I were assessed for GBA mutation detection and for their in-phase haplotype
identification, to gather genetic epidemiological data on the disease in the country and of its eventual ethnic origin. Ten missense
mutations and two complex alleles were identified. The most frequent were N370S (42.5%), L444P (20%), IVS2 + 1G > A (10%) and
R48W (5%); mutations R120W, P245H, H311R, R496H, W36X and R433G which were carried by a single chromosome each one.
Three geographical foci were identified, displaying mutation heterogeneity. N370S had multiple genetic origins, different from the
Ashkenazi’s; a single common remote ancestor for this mutation in the country was dismissed, according to the haplotype analysis.
All mutations have a likely European Caucasoid descent.

Keywords. Gaucher disease; glucocerebrosidase mutations; ethnic origins; haplotype analysis.

Introduction ‘Gaucher cells’ is typical. In the type 2 disease (OMIM:

230900) or acute neuronopathic, the patients present with
Gaucher disease (GD) (OMIM: 230800), the most common a severe hepatosplenomegaly and progressive neurologic
of lysosomal storage diseases, is an autosomal recessive deterioration being usually fatal within the first two years
disorder resulting from deficit or lack of β-glucosidase of life. Type 3 (OMIM: 231000) or chronic neuronopathic
activity. The glucocerebrosidase (acid-β glucosidase, disease has a heterogeneous presentation with variable
glucosyl-ceramidase, EC.3.2.1.45), a lysosomal glycoside neurological and visceral involvement, onset in childhood
hydrolase, cleaves the β-glucosidic linkage of glucosylce- or in adulthood; almost all patients survive until adult-
ramide, a normal intermediate of glycolipid catabolism hood (Vellodi et al. 2001).
(Beutler and Grabowski 1995). The abnormal function The gene encoding GBA (1q21) encompasses 7 kb with
leads to accumulation of the substrate (glucocerebroside) 11 exons, producing a cDNA of ∼2.5 kb; the mature
in cells of the macrophage–monocyte system. protein has 39 amino acids signal peptide. GBA has a
This disease is highly heterogeneous and has been highly homologous pseudogene (psGBA with 96% iden-
classified into three types based on the presence and rate tity), located 16-kb downstream (Horowitz et al. 1989)
of progression of neurologic manifestations. The most with the same organization of exons and introns as the
common (95% of cases) nonneuronopathic GD type 1 functional gene but carrying large deletions in introns
(OMIM: 230800) is characterized by the presence of 2, 4, 6 and 7, correspond to Alu elements. These highly
hepatosplenomegaly, pancytopenia and skeletal involve- homologous sequence and physical proximity leads to gene
ment, without neurological manifestations, and a variable conversion and crossing over events between GBA and
age of onset of symptoms; bone marrow infiltration by psGBA, producing mutated complex alleles.

583
584 Gilberto Gómez et al.

To date, in GBA gene, around 437 different mutations strand conformation polymorphism (SSCP) analysis of
have been registered in the human mutation database each exon was performed as previously reported (Paradisi
at Cardiff, UK (www.hgmd.cf.ac.uk/ac/). Nevertheless, and Arias 2010), and those showing any abnormal migra-
four mutations (N370S, L444P, 84insG, IVS2 + 1G > A) tion pattern were sequenced at Macrogen, Seoul, Korea.
account for 90% of Ashkenazim patients worldwide but
for only 50–60% of cases in different populations, many of
them being compound heterozygotes. Polymorphisms used to construct haplotypes
In this study, 20 Venezuelan independent families with
GD type I ascertained for 20 years were assessed to iden- Two intragenic single-nucleotide polymorphisms (SNP) at
tify GBA mutations and to study genetic epidemiological introns 6 and 7, g.4813G > A [rs762488; NM_000157.3:
features of the disease in the country. The most frequently c.762 − 180G > A] and g.5470G > A [NM_000157.3:
found mutation N370S had a heterogeneous origin, dif- c.999 + 240G > A], and two microsatellites (5GC3.2 and
ferent from the Ashkenazim’s according to the haplotype ITG6.2) located in the flanking regions of the gene were
analysis. used to construct the haplotypes in phase with the muta-
tions. To establish the phases, genotypes of each marker
in carriers and noncarriers family members were estab-
Materials and methods lished, and segregation analysis of the allele transmission
from parents to descendants was assessed in each poly-
Sample morphic site. Primers were previously reported by Lau
et al. (1999) and Rodríguez-Marí et al. (2001). Alleles
A total of 20 genetically unrelated index cases with at the SNPs were detected by the restriction enzymes
biochemical and clinical diagnosis of GD, who were (PvuII for g.4813G > A and Bsu36I for g.5470G >
referred to the Human Genetics Laboratory (HGL) at the A); microsatellites 5GC3.2 (dinucleotide repeats CT) and
Venezuelan Institute for Scientific Research (IVIC) and ITG6.2 (tetranucleotide repeats AAAT) were analysed in
to the Hematology Service at the Miguel Pérez Carreño a 10% and 8% polyacrylamide (acrylamide:bisacrylamide,
Hospital (Caracas) were included in this study. Two addi- 39:1) gel electrophoresis, respectively. Some samples were
tionally affected family members (a sister and an aunt) of Sanger sequenced to confirm the actual allele sizes.
one of the families (patient number 13, table 1) were also
included in the study.
The diagnosis of GD was established by the clinical Results
manifestations and a diminished β-glucosidase activity.
The enzymatic activities were measured either at the HGL, The main phenotypic and genotypic data of the studied
according to protocols by Peters et al. (1977) and Daniels index cases are shown in table 1. All patients had a type
et al. (1981), or at Centogene, Germany, using mass spec- I form of the disease (nonneuronopathic), with a highly
trometry. The geographical origin of each family was variable age of onset and common clinical manifestations.
established by recording the precise place of birth of the Almost all patients had splenomegaly (94.7%) and
remote ancestors (grandparents and great-grandparents). hepatomegaly (84.2%); more than half of them had throm-
For the molecular analyses, 5 mL blood sample was bocytopenia (52.6%) and 42.1% were anaemic. Bone crisis
collected in EDTA and DNA was extracted by saline occurred in 10.5% of cases. In 45% of patients, the diagno-
method (Lahiri and Nurnberger 1991). Written voluntary sis was made in the first decade of life, in 20% the diagnosis
informed consent was obtained from all family members was made in the second decade, and in 35% during the
according to the bioethical institutional guidelines. third, fourth or fifth decades; 14 of 20 were female. All the
patients are receiving enzymatic replacement therapy.

DNA analyses
GBA analysis
Mutation detection: A two stage polymerase chain
reaction (PCR) method was used to selectively amplify Complete coding region and intron-exon boundaries of the
the glucocerebrosidase functional gene but not the pseudo- GBA gene were screened; 10 different mutations and two
gene. In the first stage, only the functional gene was ampli- complex alleles in 40 studied chromosomes were identified,
fied in three large amplicons (between 1681 and 2971 bp). all of them previously reported in different populations.
In the second stage, the first round PCR products were used The most frequent mutation was N370S, carried by
as templates for the amplification of each and all the exons 17 chromosomes (42.5%), followed by L444P (20%), pre-
of the GBA gene (nested PCR). Primers were previously senting as a complex allele or as a single allele (5%);
reported by Stone et al. (2000). The GenBank reference IVS2+1G > A was present in 10% of chromosomes, while
sequence accession number was NM_000157.3. Single the R48W mutation was found in 5%.
 GBA mutations in Venezuelan patients 585

Mutations R120W, P245H, H311R, R496H, W36X and supporting different origins between foci for N370S. The
R433G showed low frequencies, being each one carried R48W, a very infrequent mutation worldwide, was found in
by only one chromosome out of 40. All these mutations two foci (Yaracuy and Lara states, 90 km apart: index case
cause deleterious effects on the protein structure, with numbers 2 and 3, table 1) with the same in-phase haplo-
Polyphen scores of 1.0 (P245H, H311R and R433G), 0.999 type 318; G; 222; A, suggesting a common but very remote
for R120W and 0.488 for R496H. Mutation W36X causes ancestor.
a severely truncated protein. Ninety-five per cent of index The haplotypes in phase with the 12 detected mutations
cases were compound heterozygotes. Mutated amino acid are shown in table 2. The most frequent mutation N370S
positions in all mutation names refer to the processed pro- was in phase with four different haplotypes, as well as the
tein, which does not include the 39-residue signal peptide. second most frequent L444P (alone and in complex alle-
les); in three instances, N370S and L444P had the same
haplotypes, which were also very frequent in patients: 314;
Haplotype analysis and geographic distribution G; 222; G (37.5%), 314; G; 222; A (22.5%) and 318; G;
224; G (7.5%) but not so in controls. Infrequent mutations
Only seven of 20 families (35%) had geographic had a unique in-phase haplotype, except R48W, as already
aggregation, with remote ancestors from three different mentioned.
geographic foci (Arias 1994) in Zulia state (three indepen-
dent families), in Lara state (three independent families)
and in Yaracuy state (one family); the remaining 65% fam-
ilies’ ancestors were scattered across the country (table 1). Discussion
Within the geographic foci there was mutation heterogene-
ity, with two different mutations in the Zulia state focus GD is the most frequent disorder among lysosomal
(centre in La Cañada), five mutations in the Lara state storage diseases, although it is infrequent in populations
focus (centre in Barquisimeto) and two mutations in the worldwide. In Venezuela, the ‘Venezuelan association of
Yaracuy state focus (centre in San Felipe), suggesting dif- patients with lysosomal diseases’ (AVEPEL) has 81 regis-
ferent genetic origins for GD in each one. tered independent index cases from the country at large,
In the Zulia state focus, the three unrelated index cases all receiving enzymatic replacement therapy provided by
(numbers 8, 9 and 13, table 1) were compound heterozy- the Governmental Social Security Agency. Thus, the dis-
gotes for mutations IVS2 + 1G > A and N370S, which ease prevalence in Venezuela can be estimated as 1:77000
were both in phase with the same 314; G; 222; G haplo- families if all existing cases had been detected, calculated
type in the three families. as previously reported (Paradisi et al. 2015), which is simi-
Family of index case number 13 contained two lar to the figure quoted in general worldwide populations,
compound heterozygous sisters (N370S/IVS2 + 1G > A) around 1.3:100000 inhabitants, excluding the Ashkenazi
and an affected maternal aunt carrying N370S plus Jewry of eastern and central European ancestry, in which
another nonidentified change; the paternal one (IVS2 + the prevalence is 1:500 to 1:1000 (Orphanet Reports Series,
1G > A) had a recent central European Ashkenazi ori- http://www.orpha.net).
gin but the maternal N370S frequent in the focus, is not a In 20 independent index cases, the mutation detection
Jewish mutation according to its in-phase haplotype, being rate was 92.5%. A genotype–phenotype correlation could
apparently only identical by nature (IBN). not be established, since there were 13 different geno-
Index case number 7 also carried IVS2+1G > A/N370S types, all but one index case was compound heterozy-
mutations with the same in-phase haplotypes, but her gotes for different mutations. The exception were the
remote ancestors origins were different from two far apart IVS2+1G>A/N370S carriers, which accounted for 20%
states (Sucre state and Falcón state, at the oriental and of the sample; between them, however there was no clear
occidental regions of the country). genotype–phenotype correlation (table 1).
This finding strongly suggests that in most cases, those Mutations N370S and L444P are the most common
mutations both in nonJewish and in Ashkenazi carriers mutations worldwide; jointly they represent between 50
might be likely also identical by descent (IBD), going back and 62% of the GBA mutations in all populations, except
in their ancestry much earlier than one thousand years. among the Ashkenazi, which accounts for 93% of the
In the Lara state focus, three different mutations and mutant alleles (Koprivica et al. 2000); in Venezuelan
two complex alleles were present; the three unrelated patients, its joint frequency was 62.5%. N370S is virtually
index cases (numbers 3, 4 and 6, table 1) were compound absent in Mongoloid populations.
heterozygotes [R48W]/[R120W], [N370S]/[L444P+A456P Mutation N370S has been reported to cause relatively
+V460V], and [N370S]/[E326K+L444P]. Interestingly, the minor changes in the glucocerebrosidase structure and
N370S mutation had the same in-phase 314; G; 222; A therefore, in its catalytic activity (Dvir et al. 2003). It is
haplotype, shared by the two independent families, and located at the interface of domains II and III, too far
different from that found in the Zulia state focus, thus from the active site to participle directly in catalysis. Thus,
586 Gilberto Gómez et al.

Table 1. Clinical and genetic epidemiological features of Gaucher disease index cases.

Age at diagnosis (years) In-phase

Patient Sex (years) Clinical manifestation Genotypea haplotypeb Geographic origin

1 ♀ 4 A; S; T H311R 314; G; ?; ? HGO

N370S 318; G; ?; ?
2 ♀ 2 H; S; T; Hy R48W 318; G; 222; A YSF
? 322; G; 222; G
3 ♀ 2 A; H; S; T R48W 318; G; 222; A LSF
R120W 326; G;222, G
4 ♂ 9 A; H; T N370S 314; G; 222; A LSF
RecNciI 314; G; 222; A
5 ♂ 33 NA P245H 314; G; 222; G HGO
N370S 314; G; 222; G
6 ♀ 27 A; H; S; T; Bc N370S 314; G; 222; A LSF
[E326K+L444P] 318; G; 222, G
7 ♀ 2 H; S; T IVS2 + 1G > A 314; G; 222; G HGO
N370S 314; G; 222; G
8 ♂ 24 H; S IVS2 + 1G > A 314; G; 222; G ZSF
N370S 314; G; 222; G
9 ♀ 8 A; H; S; T IVS2 + 1G > A 314; G; 222; G ZSF
N370S 314; G; 222; G
10 ♂ 2 A; H; S; T R433G 318; G; 224; A HGO
L444P 318; G; 224; G
11 ♀ 4 H; S; T N370S 314; G, 222; G HGO
L444P 318; G; 222;G
12 ♀ 11 A; H; S N370S 318; G; 224; G HGO
? 318; G; 224; G
13 ♀ 8 S; T IVS2 + 1G > A 314; G; 222; G ZSF
N370S 314; G; 222; G
14 ♀ 56 H; S N370S 314; G; 222; A HGO
? 314; G; 222; A
15 ♀ 50 H; S; Bc RecNciI 314; G; 222; A HGO
R496H 318; G; 222; A
16 ♂ 9 A; H; S N370S 314; G; 222; A HGO
RecNciI 318; G; 222; G
17 ♀ 21 H; S W36X 314; G; 222; G HGO
N370S 314; G; 222; G
18 ♂ 44 S N370S 314; G; 222; G HGO
N370S 314; G; 222; G
19 ♀ 13 H; S N370S 314; G; 222; A HGO
RecNciI 314; G; 222; G
20 ♂ 18 H; S N370S 314; G, 222; A HGO
RecNciI 318; G; 222; G

A, anemia; H, hepatomegaly; S, splenomegaly; T, thrombocytopenia; Bc, bone crisis; Hy, hypotonia; NA, data not available;
RecNciI: complex allele [L444P+A456P+V460V]; ‘?’ unidentified genotype; HGO, heterogeneous geographic origins; ZSF, Zulia
state focus; YSF, Yaracuy state focus; LSF, Lara state focus.
a Mutation nomenclature refers to the processed protein, not including the 39 residues of the signal peptide.
b Haplotype markers from left to right: ITG6.2; g.5470G > A (c.999 + 240G > A); 5GC3.2; g.4813G > A (c.762 − 180G > A).

its phenotypic effect tends to be moderate, producing 1990; Amaral et al. 1997; Cormand et al. 1998; Rockah
always type I disease with mild clinical manifestations and et al. 1998), as well as with the A allele at intron 6
an older age at diagnosis, even when in heterozygosity (g.4813G > A, which suppress the restriction site for
(Charrow et al. 2000), as was observed in our patients the PvuII enzyme known as the Pvu1.1− haplotype); and
(table 1). Polyphen analysis classifies its pathogenicity as the G allele at g.5470G > A (c.999 + 240G > A).
possibly damaging, with a score of 0.607. Thus, haplotype 318; G; 222; A (ITG6.2; g.5470G > A;
As mentioned, mutation N370S is the most frequent 5GC3.2; g.4813G > A) is almost always in phase with the
in Ashkenazi due to a founder effect and a genetic drift Ashkenazi N370S mutation. In Venezuelan patients, the
phenomena. In such populations, it shows a strong allelic N370S had at least three different in-phase haplotypes:
disequilibrium with the 222 bp and 318 bp alleles of 314; G; 222; G (47.1%), 314; G; 222; A (35.3%), 318; G;
microsatellite markers 5GC3.2 and ITG6.2 (Zimran et al. 224; G (11.8%) and 318; G; (-); (-) (5.8%) (table 2). The
 GBA mutations in Venezuelan patients 587

Table 2. GBA mutations, in-phase haplotypes and its frequencies in patients and in
a control sample.

Haplotype frequencies (%)

GBA mutation In-phase haplotype Patients (n = 40) Controls (n = 56)

N370S 314; G; 222; A 22.5 14.3

RecNciI*
R48W 318; G; 222; A 7.5 26.8
R496H
L444P 318; G; 222; G 12.5 41.1
[E326K/L444P]*
RecNciI*
P245H 314; G; 222; G 37.5 16.1
W36X
RecNciI*
IVS2 + 1G > A
N370S
Unidentified 322; G; 222; G 2.5 1.8
N370S 318; G; 224; G 7.5 0
L444P
R433G 318; G; 224; A 2.5 0
R120W 326; G; 222; G 2.5 0
N370S 318; G; (-); (-) 2.5 0
H311R 314; G; (-); (-) 2.5 0

Some haplotypes were found to be in phase with more than one mutation type, and
some mutations had more than one in-phase haplotypes (N370S, L444P). Haplotype
frequencies were different between patients and control individuals; the most frequent
in controls (318; G; 222; G) were not in patients chromosomes, suggesting that some
mutations arose in a different genetic background. Some in-phase haplotypes were
not found in the control sample. n, Number of chromosomes; ‘-’ uninformative (n = 2
chromosomes); ‘*’ complex alleles; RecNciI, L444P+A456P+V460V.
Haplotype markers from left to right: ITG6.2; g.5470G > A (c.999 + 240G > A);
5GC3.2; g.4813G > A (c.762 − 180G > A).

in-phase haplotype with the highest frequency (314; G, N370S) in Ashkenazi, Caucasoids (Stinermann et al. 2012)
222; G), has also been found by Wilches et al. (2006) in and also in the Venezuelan studied patients, but not in east
Colombian N370S carriers; Colombian and Venezuelan Asians, Caucasoids or sub-Saharan Africans.
populations share similar demographic history, suggest- Two haplotypes in phase with the L444P mutation as
ing a plausible common remote origin for the mutation a single allele (318; G; 222; G and 318; G; 224; G),
carrying this haplotype in phase. On the other hand, hap- and two other in complex alleles (table 2) were found in
lotypes in phase with this N370S ‘Venezuelan’ mutation the index cases. Tuteja et al. (1993) demonstrated that
suggest that the origin is different from the Ashkenazi’s in several haplotypes are in phase with this mutation and
most cases, and that there is not a single common ancestor suggested multiple ancestral origins for it, since codon 444
for the mutation in the country. In Venezuela, most of the could be a hot spot for mutations. The mutation has been
N370S alleles are distributed in the northwestern region, associated with the Pv1.1− haplotype in Jewish popula-
showing geographic aggregation in two foci: La Cañada tions (Rodríguez-Marí et al. 2001). In Venezuelan patients,
(Zulia state) and Barquisimeto (Lara state). Nevertheless, all chromosomes carrying L444P were in phase with
in-phase haplotypes were different between foci (314; G; allele G (haplotype Pv1.1+ ), thus discarding a possible
222; G and 314; G; 222; A) respectively, suggesting sepa- remote Jewish origin for the mutation. Further, haplo-
rate origins for the N370S in each focus, but a common type diversity suggests multiple origins of L444P in the
source within each one. country.
The L444P occurs in the hydrophobic core of the Mutation IVS2 + 1G > A was identified in four patients
Ig-like domain (domain II), causing protein instability due (four chromosomes, 10%). The change produces aber-
to disruption of the hydrophobic core and altered folding rantly spliced mRNAs, missing exon 2, and severely
of the domain (Beutler and Kuhl 1986; Dvir et al. 2003); impaired catalytic activity (He and Grabowski 1992). This
its Polyphen score is 0.938, suggesting a strong mutation mutation represents 1–4% of patients with GD, in the
effect on the protein structure. This mutation in homozy- ‘general’ population, ordinarily associated to the N370S
gous condition produces the chronic neuronopathic form mutation, as was seen in our patients. In all instances,
of GD. It is the second most frequent mutation (after IVS2 + 1G>A was in phase with the same haplotype
588 Gilberto Gómez et al.

314; G; 222; G. Three of the four alleles were found in Scriver and A. Beaudet), 7th edition, pp. 2641–2670. Mc Graw-
the same geographic focus (La Cañada, Zulia state); thus Hill, New York, USA.
haplotype sharing and geographic aggregation strongly Cormand B., Harboe T., Gort L., Campoy C., Blanco M.
and Chamoles N. 1998 Mutation analysis of Gaucher dis-
suggest a common remote ancestor and a founder effect ease patients from Argentina: High prevalence of the RecNciI
as well, for these families in the country. mutation. Am. J. Med. Genet. 80, 343–351.
Mutation R48W had only one in-phase haplotype found Charrow J., Andersson H., Kaplan P., Kolodny E., Mistry P., Pas-
in two unrelated patients from different, although con- tores G. et al. 2000 The Gaucher registry: demographics and
tiguous geographic foci (in Lara and Yaracuy states). The disease characteristics of 1698 patients with Gaucher disease.
Arch. Intern. Med. 160, 2835–2843.
mutation is very infrequent worldwide (Choy et al. 2007), Choy F., Zhang W., Hui-Ping S., Zay A., Campbell T., Tang N.
thus haplotype sharing might suggest a common and very and Ferreira P. 2007 Gaucher disease among Chinese patients:
remote origin for it in the northern occidental area of the review on genotype/phenotype correlation from 29 patients
country. R48W has a severe effect on the protein, as is and identification of novel rare alleles. Blood Cells Mol. Dis. 38,
suggested by its Polyphen score of 1.0. 287–293.
Daniels L., Glew W., Diven R., Lee E. and Radin N. 1981
Patients with GD in Venezuela are geographically An improved fluorometric leucocyte β-glucosidase assay for
widespread, and their parents typically are not closely Gaucher’s disease. Clin. Chim. Acta 115, 369–375.
related, but three geographic foci were detected for 35% Dvir H., Harel M., Mc Carthy A., Toker L., Silman I., Futerman
of the families. This suggests both remote kinship and a H. and Sussman J. 2003 X-ray structure of human acid-β-
relatively high frequency of diverse mutant alleles, beside a glucosidase, the defective enzyme in Gaucher disease. EMBO
Rep. 4, 704–710.
founder phenomenon. There was great genetic heterogene-
He G. S. and Grabowski G. 1992 Gaucher disease: A G+1 →
ity of mutations, with most of the patients being compound A+1 IVS2 splice donor site mutation causing exon 2 skipping
heterozygotes as is the usual finding everywhere, except in the acid β-giucosidase mRNA. Am. J. Hum. Genet. 51, 810–
among the Ashkenazi. However three mutations account 820.
for 72.5% of the chromosomes: N370S (42.5%), L444P Horowitz M., Wilder S., Horowitz Z., Reiner O., Gelbart T. and
(20%), and IVS2+1G>A (10%). Thus, in any new patient, Beutler E. 1989 The human glucocerebrosidase gene and pseu-
dogene: structure and evolution. Genomics 4, 87–96.
these mutations should be the first tested in our popu- Koprivica V., Stone D., Park J., Callahan M., Frisch A. and
lations, being they also the most common in European Cohen I. 2000 Analysis and classification of 304 mutant alle-
Caucasoids from diverse geographic regions, but not so in les in patients with type 1 and type 3 Gaucher disease. Am. J.
other ethnic groups. Hum. Genet. 66, 1777–1786.
Lahiri D. and Nurnberger J. 1991 A rapid non-enzymatic method
for the preparation of HMW DNA from blood for RFLP stud-
Acknowledgements ies. Nucleic Acids Res.19, 5444.
Lau E., Tayebi N., Ingraham L., Winfield S., Koprivica V., Stone
To all the patients who generously gave their consent to D. et al. 1999 Two novel polymorphic sequences in the gluco-
participate in the study. To Mrs Joelkys Romero and Mrs cerebrosidase gene region enhance mutational screening and
Nereyda Guédez, president and vice-president of the Venezue- founder effect studies of patients with Gaucher disease. Hum.
lan Association of patients with lysosomal diseases (AVEPEL) Genet. 104, 293–300.
for their support. To Lic. Neida González for the biochemical Paradisi I. and Arias S. 2010 Marked geographic aggrega-
processing of the samples at the Human Genetics Laboratory, tion of acute intermittent porphyria families carrying muta-
IVIC. tion Q180X in Venezuelan populations, with description
of further mutations. J. Inherit. Metab. Dis. 33, suppl 3,
S455–S463.
Paradisi I., De Freitas L. and Arias S. 2015 Most frequent muta-
References tion c.3402delC (p.Ala1135GlnfsX13) among Wilson disease
patients in Venezuela has a wide distribution and two old ori-
gins. Eur. J. Med. Genet. 58, 59–65.
Arias S. 1994 Selective detection among the high risk popula- Peters S., Coffee R., Glew R., Lee D., Wenger S. and Li Y. 1977
tions, instead of mass screening, proposed for countries with Isolation of heat-stable glucocerebrosidase activators from the
various homogeneous origins and a stratified distribution of spleens of three variants of Gaucher disease. Arch. Biochem.
abnormal genes. In New horizons in neonatal screening (ed. J. Biophys. 183, 290–297.
P. Farriaux and J. L. Dhondt), Int. congr ser, vol. 1041, pp. 97– Rockah R., Narinsky R., Frydman M., Cohen I., Zaizov R.,
180. Excerpta media, Amsterdam. Weizman A. and Frisch A. 1998 Linkage disequilibrium of
Amaral O., Marcao A., Pinto E., Zimran A. and Sa Miranda M. common Gaucher disease mutations with a polymorphic site
1997 Distinct haplotype in non-Ashkenazi Gaucher disease in the Pyruvate Kinase (PKLR) gene. Am. J. Med. Genet. 78,
patients with N370S mutation. Blood Cells Mol. Dis. 23, 416– 233–236.
425. Rodríguez-Marí A., Díaz-Font A., Chabás A., Pastores G., Grin-
Beutler E. and Kuhl W. 1986 Glucocerebrosidase processing in berg D. and Vilageliu L. 2001 New insights into the origin
normal fibroblasts and in fibroblasts from patients with type of the Gaucher disease-causing mutation N370S: extended
I, type II and type III Gaucher disease. Proc. Natl. Acad. Sci. haplotype analysis using the 5GC3.2, 5470G/A, and ITG6.2
USA 83, 7472–7474. polymorphism. Blood Cells Mol. Dis. 27, 950–959.
Beutler E. and Grabowski G. 1995 Gaucher disease. In The Stirnemann J., Vigan M., Hamroun D., Heraoui D., Rossi-
metabolic and molecular bases of inherited disease (ed. C. R. Semerano L., Berger M. et al. 2012 The French Gaucher’s
 GBA mutations in Venezuelan patients 589

disease registry: clinical characteristics, complications and force of the European working group on Gaucher diseases.
treatment of 562 patients. Orphanet. J. Rare Dis. 7, 1172–1177. Management of neuronopathic Gaucher disease: a European
Stone D., Tayebi N., Orvisky E., Stubblefield B., Madike V. consensus. J. Inherit. Metab. Dis. 24, 319–327.
and Sidransky E. 2000 Glucocerebrosidase gene mutations in Wilches R., Vega H., Echeverri O. and Barrera L. 2006
patients with type 2 Gaucher disease. Hum. Mutat. 15, 181– Los haplotipos colombianos de la mutación N370S cau-
188. sante de la enfermedad de Gaucher pueden provenir
Tuteja R., Bembi B., Agosti E. and Baralle F. 1993 1448C muta- de un haplotipo ancestral común. Biomédica 26,
tion linked to the Pv1.1− genotype in Italian patients with 434–441.
Gaucher disease. Hum. Mol. Genet. 2, 781–784. Zimran A., Gelbart T. and Beutler E. 1990 Linkage of the PvulI
Vellodi A., Bembi B., de Villemeur T., Collin-Histed T., Erickson polymorphism with the common Jewish mutation for Gaucher
A., Mengel E. et al. 2001 Neuronopathic Gaucher disease task disease. Am. J. Hum. Genet. 46, 902–905.

Corresponding editor: Kunal Ray