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International Journal of Neuroscience
ISSN: 0020-7454 (Print) 1543-5245 (Online) Journal homepage: http://www.tandfonline.com/loi/ines20
Association of VDR gene polymorphisms with risk
of Relapsing-remitting multiple sclerosis in an
Iranian Kurdish population
Rasoul Abdollah zadeh, Parisa Moradi Pordanjani, Farideh Rahmani,
Fatemeh Mashayekhi, Asaad Azarnezhad & Yaser Mansoori
To cite this article: Rasoul Abdollah zadeh, Parisa Moradi Pordanjani, Farideh Rahmani,
Fatemeh Mashayekhi, Asaad Azarnezhad & Yaser Mansoori (2017): Association of VDR gene
polymorphisms with risk of Relapsing-remitting multiple sclerosis in an Iranian Kurdish population,
International Journal of Neuroscience, DOI: 10.1080/00207454.2017.1398158
To link to this article: http://dx.doi.org/10.1080/00207454.2017.1398158
Accepted author version posted online: 26
Oct 2017.
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Download by: [University of Florida]
Date: 27 October 2017, At: 13:58
Publisher: Taylor & Francis
Journal: International Journal of Neuroscience
DOI: https://doi.org/10.1080/00207454.2017.1398158
Downloaded by [University of Florida] at 13:58 27 October 2017
Association of VDR gene polymorphisms with risk of Relapsing-remitting
multiple sclerosis in an Iranian Kurdish population
Rasoul Abdollahzadeha, b, Parisa Moradi Pordanjanic, Farideh Rahmanid, Fatemeh Mashayekhie,
Asaad Azarnezhadf, b*, Yaser Mansooria, b*
a Noncommunicable Diseases Research Center, Fasa University of Medical Sciences, Fasa, Iran
b Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
c Department of biology, Faculty of Science, Alzahra University, Tehran, Iran
d Department of Medical Biotechnology, School of Medicine, Hamedan University of Medical Sciences, Hamedan, Iran
e Department of Cell & Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran
f Cellular and Molecular Research Center, Kurdistan University of Medical Sciences, Sanandaj, Iran
Running head: VDR gene polymorphisms and multiple sclerosis
* Correspondence
Asaad Azarnezhad
Cellular and Molecular research center, School of Medicine, Kurdistan University of Medical
Sciences, Sanandaj, Iran
Azarnezhad@gmail.com
* Co-correspondence
Yaser Mansoori
Noncommunicable Diseases Research Center, Fasa University of Medical Sciences, Fasa, Iran
fums.mansoori@gmail.com
** First and second authors have an equal contribution to the work.
Abstract
Vitamin D receptor (VDR) polymorphisms have been reported to be associated with multiple
sclerosis (MS). The purpose of this study was to evaluate the association of VDR Apa-I, Bsm-I,
Fok-I, Taq-I polymorphisms with MS risk in an Iranian Kurdish population. A population
including of 118 patients affected with MS and 124 healthy matched controls were recruited to
the study. DNA was extracted from blood and genotyping of single nucleotide polymorphisms
(SNPs) was accomplished using polymerase chain reaction–restriction fragment length
polymorphism (PCR–RFLP). Fok-I and Taq-I showed significant association with risk of MS (P
< 0.05). The frequency of allele T of Fok-I (P = 0.003) and allele C of Taq-I (P = 0.0003) was
significantly different between case and control subjects and showed significant association with
risk of MS (OR = 1.84, 95% CI = 1.23-2.76; OR= 1.98, 95% CI = 1.36-2.87, respectively). CT
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genotype (OR = 1.7, 95% CI = 1.05-2.99) of Fok-I and CC genotype (OR = 2.18, 95% CI =
1.05-4.52) of Taq-I showed a predisposing effect. Combined TT+TC vs. CC for Fok-I (OR
=2.15, 95% = CI 1.29-3.60) and combined CC+TC vs. TT for Taq-I (OR = 2.58, 95% CI 1.514.40) were susceptibility genotypes for MS. Apa-I and Bsm-I were not significantly associated
with risk of MS (OR < 1, P > 0.05) and any genotypes in any genetic models were not
significantly different between cases and controls (P > 0.05). As a result, Fok-I and Taq-I
showed significant association with risk of MS, while Apa-I and Bsm-I were not observed to be
related to the risk of the disease in this population.
Keywords: Multiple Sclerosis, Genetic Variation, Single Nucleotide Polymorphism, Association Study,
Vitamin D Receptor, Kurdish Population
1. Introduction
MS is known as a chronic autoimmune inflammatory disease characterized by demyelination of
different areas of the central nervous system (CNS). It is the second most common neurological
disability which is occurred among young adult with the sex ratio of 2: 1 (women: men) [1].
According to the latest report by Iran's Ministry Of Health and Medical Education (MOHME) in
2011, MS prevalence was about 45 in 100,000 in which 77% of patients were women and 70%
of cases were in the age range of 20-40 years [2, 3].
Although the etiology of MS is still not clear, genetic, epidemiological, and immunogenetics
studies have characterized it as a multifactorial autoimmune disease [4]. Environmental factors
such as viral infections, obesity, smoking and low levels of vitamin D or its metabolites have
2
reported being involved in the pathogenesis of MS [5, 6]. Our body acquires vitamin D either
through diet or exposure to sunlight. The main metabolite of vitamin D is hydroxylated in the
liver to produce 25-monohydroxyvitamin D [25 (OH) D] which is then converted to bioactive
1,25- dihydroxy vitamin D [1,25 (OH) 2D] in the kidney. This bioactive molecule act as a ligand
for vitamin D (VDR) nuclear receptor that consequently leads to the initiation of related
biological responses [7]. Correlation of vitamin D with the increased risk of MS development
has previously been reported. The low levels of vitamin D are associated with the disease and
lower levels of vitamin D has been observed in relapse than remission phase of the disease [8].
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Therefore, the incorrect action and deficient levels of vitamin D could influence on the
development of MS.
VDR gene is located on chromosome 12q13.1 and has 9 exons and 8 introns. The promoter
region is located in exon 1 and controls the production of tissue-specific transcripts [9]. Second
and third exons encode DNA binding domain, while exons 6-9 encodes 1, 25 (OH) 2D binding
domain. One goal of human genetics is to find the genetic etiology of common and complex
disease such as MS. Genome-wide association study (GWAS) has played a considerable role in
this area. The output of GWAS is many single SNPs that will need to be confirmed in different
independent case-control studies in different populations [10]. Entrez SNP database has
published more than 30 SNPs in VDR gene from which the association of four SNPs with
autoimmune diseases such as MS has been widely studied. These SNPs including ApaI
(rs7975232), BsmI (rs1544410), FokI (rs10735810), and TaqI (rs731236) [11].
Clear conclusions have not been obtained about the possible role of VDR polymorphisms in
susceptibility to MS and achieved data have been conflicting. Various factors, including
methodological challenges, genetic or ethnic differences in studied populations and various
environmental factors could explain these discrepancies. Therefore, we encouraged to design a
case-control study aim to assess the association of ApaI, BsmI, FokI, and TaqI polymorphisms
with risk of MS in an Iranian Kurdish population.
2. Materials and Methods
2.1 Study population
3
A study population containing 118 patients affected with MS confirmed based on McDonald
criteria and 124 matched healthy subjects without any family history of neurologic and systemic
disorders were recruited to this study. All procedures performed in accordance with the ethical
standards of the institutional and/or national research committee and with the 1964 Helsinki
Declaration and its later amendments or comparable ethical standards. Informed consent form
was obtained from all the participants included in the study.
2.2 DNA extraction and genotyping
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Five ml blood sample was taken from each of participants and genomic DNA extraction was
performed according to a standard salting-out protocol with a little modification [12]. All the
procedures consisting of primers sequences, PCR-RFLP, and genotyping were according to our
previously published protocol [13]. Briefly, target regions were PCR-amplified and digestion of
PCR products was performed with specific restriction enzymes including ApaI, BsmI, FokI, and
Taq. Genotyping was accomplished by running of digested products on a 2.5% agarose gel
electrophoresis. Genotype patterns of selected VDR polymorphisms are illustrated in Figure 1.
2.3 Statistical analysis
Deviation from Hardy-Weinberg Equilibrium was assessed by calculating of allelic and genotype
distribution between the case and the control subjects. Differences in allelic and genotype
frequency between MS patients and healthy controls was evaluated by chi-square (χ2) and
independent sample T-tests. Descriptive statistics were reported as the mean± standard deviation.
P < 0.05 was considered as the statistically significant level. Odds ratio (OR) and 95%
confidential interval (95% CI) were determined for genetic risk of each allele and genotype. All
the analyses were done using SPSS Software version 16.0 (Chicago, IL).
3. Results
3.1 Study population
This study was accomplished on 118 MS patients and 124 healthy matched subjects. Statistical
Chi- square and t-test showed no significant difference between case (P = 0.874) and control (P =
0.319) groups in sex ratio. The MS patients have the same age range and sex ration compared
with controls. Characteristics, as well as inclusion and exclusion criteria of the MS patients and
4
control participants are summarized in Table 1. Case and control groups were in HardyWeinberg equilibrium for each of SNPs (Table 2).
3.2 Association of ApaI, BsmI, FokI and TaqI polymorphisms with MS
The genotype and allele-frequencies of ApaI, BsmI, FokI and TaqI polymorphisms in the MS
and control individuals are described in Table 2. As it is shown, there are significant differences
in genotype and allele frequencies between patients and controls for SNP FokI (rs2228570) (P =
0.003) in which T allele found to be positively associated with risk of the disease (OR = 1.84,
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95% CI = 1.23- 2.76), while C allele showed a negative correlation (OR = 0.54, 95% CI = 0.360.81). Genotypes of CC (OR = 0.46, 95% CI = 0.28- 0.78) and CT (OR = 1.7, 95% CI = 1.052.99) revealed to be significantly correlated with the disease and were higher in cases than
controls (P = 0.004, P = 0.032; respectively), whereas no significant difference was observed
between the case and control subjects for TT genotype (P = 0.136). According to Table 3, a
significant difference between patients and healthy subjects was observed in the recessive
genetic model (OR = 2.15, 95% CI = 1.29- 3.60). The dominant genetic model was not
significantly different between two groups (P = 0.136).
For TaqI SNP (rs731236), highly significant differences in allele frequencies were found
between patients and controls in which allele T revealed a negative association (OR = 0.51, 95%
CI = 0.39- 0.73) and allele C showed a positive correlation (OR = 1.98, 95% CI = 1.36- 2.87).
Odds ratio of TT genotype was OR= 0.39, 95% CI = 0.23- 0.66 and for carriers of genotype CC
was OR= 2.18, 95% CI = 1.05- 4.52. The TC genotype was not observed significantly different
between two groups (P =0.059). Using dominant and recessive genetic models, significant
difference was observed for both recessive model (P = 0.0005) and dominant model (P = 0.036).
In the current study, significant differences in the genotype and allele frequencies of ApaI
(rs7975232) and BsmI (rs1544410) polymorphisms were not observed among patients affected
with MS and healthy controls (P > 0.05). Similarly, significant differences between the control
and healthy subjects were not observed for any dominant or recessive genetic models (OR < 1)
(Table 3).
4. Discussion and conclusion
5
Strong pieces of evidence have shown that vitamin D status can play a vital role in susceptibility
to and progress of MS. These evidences include (i) correlation of reduced levels of 25 (OH) D in
plasma with increased risk of susceptibility to MS [14], (ii) low prevalence of disease in the
tropics and high prevalence in high latitude climates [15], (iii) relation of seasonal variations
with variations in 25 (OH) D levels [16], (iv) lower bone mass density (BMD) in patients with
MS [17], and (v) Genetic variants in several genes involved in metabolism and activity of
vitamin D, especially VDR gene [13, 18, 19].
Association of VDR gene polymorphisms with MS have been conducted in different
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transmission and case- control studies. To the best of authors’ knowledge, the present work is the
first report performed on Kurdish population aimed to examine the correlation of four common
VDR gene polymorphisms with MS risk. Iran is an ethnically heterogeneous country and
therefore the association of these polymorphisms with MS may be substantially varied between
Iranian populations. Thus far, many studies have been conducted on the association of VDR gene
polymorphisms with MS but with contradictory conclusions. Ethnicity differences, geographical
diversity, interaction with other genetic or environmental factors and clinical heterogeneity can
be the reasons for these inconsistencies.
ApaI (A / C), BsmI (A / G), and TaqI (T / C) are located near to the 3’ end of VDR gene, while
FokI polymorphism is located near to the 5 ' end. It is reported that ApaI and BsmI (located in
intron 8) and TaqI (located in exon 9) have no effects on VDR protein structure, while FokI
(located in exon 2) results in a longer VDR protein with three more amino acids through the
creation of an initiation codon. Additionally, ApaI, BsmI, and TaqI variants have been found to
be in a strong linkage disequilibrium (LD) [9]. Although these SNPs do not cause structural or
protein expression changes, being in LD with other variants in the 3-UTR of VDR gene may
influences on VDR transcript stability by introducing changes in miRNA binding sites or other
regulatory factors and gene expression.
FokI polymorphism creates a new start codon in exon 2 which leads to two types of proteins with
427 amino acids (f- Allele) and 424 amino acids (normal alleles; F- Allele). There are conflicting
results regarding the different expression activity between these isoforms [20, 21]. However,
there is a more effective interaction between F isoform and transcription factor co-activator IIB
(TFIIB) than f isoform [22]. It has also reported that there is a functional effect of this
6
polymorphism on immune responses [23]. In the current study, significant differences in
genotype and allele frequencies of FokI polymorphism (excluding of homozygous TT) were seen
between patient and control groups. T allele frequency in patients and controls was
approximately 34% and 22%, respectively, which means that allele T has a positive or
predisposing association with MS. On the other side, CC genotype showed a protective role
against the development of disease, while CT genotypes found to increases risk of MS. However,
no significant difference was observed between patients and healthy subjects for TT genotype.
Assuming the recessive genetic model for this SNP, a significant difference was observed for
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combined TT + TC versus CC between the control group and patients. Many studies have
confirmed our results and the association of FokI and the risk of MS have been reported [24-26].
However, there are other reports showing no association between this variant and MS [13, 2729]. There are more than 30 VDR polymorphisms from which FokI is associated with levels of
25 (OH) D. Many kinds of literatures have shown that allele F is responsible for lower levels of
25 (OH) D. It could be concluded that this variant may affect vitamin D metabolism and its
metabolite levels, especially 25 (OH) D, and thereby change the rate of progression and severity
of the disease [24, 30].
TaqI was the second studied polymorphisms in which significant differences in genotype and
allele frequencies (except heterozygous TC) was observed between cases and controls. Allele C
showed a positive association with MS and its frequency was higher in patients (46.2%) than
controls (30.2%). As it was expected, TT genotype showed a negative correlation with the
disease, while the CC genotype revealed to be positively associated with MS. Although no
significant difference was observed between patients and controls for TC genotype, it was close
to significant value. Considering a dominant genetic model, combined TT + TC vs CC found to
be significantly different between two groups with a protective role against the disease, however,
combined CC + TC compared to TT genotype was a susceptibility recessive genetic model
associated with risk of MS. There are several studies in consistent with our findings [13, 19, 26,
27, 31] nonetheless, the other reports are in contrast with ours [18, 24, 25, 32, 33]. These
findings explain that allele T and TT genotype seem to be directly or indirectly by LD with other
variants introduce susceptibility risk to MS, however, carriers of allele C and CC genotype might
be more protected against the disease.
7
ApaI was another SNP that was not significantly associated with MS neither protective nor
predisposing. According to the demonstrated results, no significant differences either in allele or
genotype frequencies were found between MS and healthy control subjects. Regarding dominant
and recessive genetic models, any significant differences were also not observed. There are many
kinds of studies conducted on the relation of this SNP with MS which have shown conflicting
results [13, 25, 27, 34]. Therefore, lack of significant association might be due to population
stratification, inter-population genetic variation, false positive studies, false negative studies or
true variability in association among different populations, small sample size, and need to greater
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statistical power. Our data did not also show significant association between BsmI and MS.
Allele and genotype frequencies and any of assumed genetic models were not significantly
differences between patients and controls. This result has been confirmed in some reports but not
all [11, 13, 24, 35]. Lack of association might be explained by the same limitation which
mentioned for ApaI.
In conclusion, our evidence disclosed a significant association between VDR Taq-I and Fok-I
polymorphisms and MS risk, however, our data do not support any significant correlations
between BsmI and ApaI polymorphisms and risk of MS. To be able to speak definitively about
the findings, a study of larger population and functional assays are needed. Assessing the effect
of the selected SNPs in the current study and other VDR polymorphisms on MS is also suggested
to be performed in different ethnical population.
Acknowledgment
We would like to show our gratitude to the patient and healthy individuals for participating in
this work and providing us the complementary information. This research was financially
supported by Fasa University of Medical Sciences under Grant 95125.
Conflict of interest
All the authors declare that there is no conflict of interest.
References
8
1.
Weiner, H.L., Multiple sclerosis is an inflammatory T-cell–mediated autoimmune disease. Arch
Neurol, 2004. 61(10): p. 1613-1615.
2.
Izadi, S., et al., Significant increase in the prevalence of multiple sclerosis in iran in 2011. Iran J
Med Sci, 2014. 39(2): p. 152.
3.
Sahraian, M.A., et al., Multiple sclerosis in Iran: a demographic study of 8,000 patients and
changes over time. Eur. Neurol, 2010. 64(6): p. 331-336.
4.
Abdollahzadeh, R., et al., Polymorphisms of RPS6KB1 and CD86 associates with susceptibility to
multiple sclerosis in Iranian population. Neurol. Res, 2017. 39(3): p. 217-222.
5.
Ascherio, A. and K.L. Munger, Environmental risk factors for multiple sclerosis. Part I: the role
Downloaded by [University of Florida] at 13:58 27 October 2017
of infection. ‎Ann. Neurol, 2007. 61(4): p. 288-299.
6.
Ascherio, A. and K.L. Munger, Environmental risk factors for multiple sclerosis. Part II:
Noninfectious factors. ‎Ann. Neurol, 2007. 61(6): p. 504-513.
7.
Huang, J. and Z.-F. Xie, Polymorphisms in the vitamin D receptor gene and multiple sclerosis
risk: a meta-analysis of case–control studies. ‎J. Neurol. Sci, 2012. 313(1): p. 79-85.
8.
Ascherio, A., K.L. Munger, and K.C. Simon, Vitamin D and multiple sclerosis. Lancet Neurol,
2010. 9(6): p. 599-612.
9.
Uitterlinden, A.G., et al., Genetics and biology of vitamin D receptor polymorphisms. Gene,
2004. 338(2): p. 143-156.
10.
Bush, W.S. and J.H. Moore, Genome-wide association studies. PLoS Comput Biol, 2012. 8(12):
p. e1002822.
11.
Smolders, J., et al., The relevance of vitamin D receptor gene polymorphisms for vitamin D
research in multiple sclerosis. Autoimmun Rev, 2009. 8(7): p. 621-626.
12.
MWer, S., D. Dykes, and H. Polesky, A simple salting out procedure for extracting DNA from
human nucleated cells. Nucleic acids res, 1988. 16: p. 1215.
13.
Abdollahzadeh, R., et al., Predisposing role of vitamin D receptor (VDR) polymorphisms in the
development of multiple sclerosis: A case-control study. ‎J. Neurol. Sci, 2016. 367: p. 148-151.
14.
van der Mei, I.A., et al., Vitamin D levels in people with multiple sclerosis and community
controls in Tasmania, Australia. J. Neurol, 2007. 254(5): p. 581.
15.
Kurtzke, J., Epidemiology of multiple sclerosis. Does this really point toward an etiology? Lectio
Doctoralis. ‎Neurol. Sci, 2000. 21(6): p. 383-403.
16.
Ogawa, G., et al., Seasonal variation of multiple sclerosis exacerbations in Japan. ‎Neurol. Sci,
2004. 24(6): p. 417-419.
9
17.
Ozgocmen, S., et al., Vitamin D deficiency and reduced bone mineral density in multiple
sclerosis: effect of ambulatory status and functional capacity. J Bone Miner Metab, 2005. 23(4):
p. 309-313.
18.
Smolders, J., et al., Association study on two vitamin D receptor gene polymorphisms and vitamin
D metabolites in multiple sclerosis. Ann. N. Y. Acad. Sci, 2009. 1173(1): p. 515-520.
19.
Bermúdez-Morales, V.H., et al., Vitamin D receptor gene polymorphisms are associated with
multiple sclerosis in Mexican adults. ‎J. Neuroimmunol, 2017.
20.
Gross, C., et al., The vitamin D receptor gene start codon polymorphism: a functional analysis of
FokI variants. J Bone Miner Metab, 1998. 13(11): p. 1691-1699.
Downloaded by [University of Florida] at 13:58 27 October 2017
21.
Arai, H., et al., A vitamin D receptor gene polymorphism in the translation initiation codon: effect
on protein activity and relation to bone mineral density in Japanese women. J Bone Miner Metab,
1997. 12(6): p. 915-921.
22.
Jurutka, P.W., et al., The polymorphic N terminus in human vitamin D receptor isoforms
influences transcriptional activity by modulating interaction with transcription factor IIB. ‎Mol.
Endocrinol, 2000. 14(3): p. 401-420.
23.
van Etten, E., et al., The vitamin D receptor gene FokI polymorphism: functional impact on the
immune system. ‎Eur. J. Immunol, 2007. 37(2): p. 395-405.
24.
Tizaoui, K., et al., Association between vitamin D receptor polymorphisms and multiple sclerosis:
systematic review and meta-analysis of case–control studies. Cell Mol Immunol, 2015. 12(2): p.
243-252.
25.
Simon, K.C., et al., Polymorphisms in vitamin D metabolism related genes and risk of multiple
sclerosis. ‎Mult. Scler. J, 2010. 16(2): p. 133-138.
26.
Al-Temaimi, R.A., et al., The association of vitamin D receptor polymorphisms with multiple
sclerosis in a case-control study from Kuwait. PloS one, 2015. 10(11): p. e0142265.
27.
Tajouri, L., et al., Variation in the vitamin D receptor gene is associated with multiple sclerosis in
an Australian population. J. Neurogenet, 2005. 19(1): p. 25-38.
28.
Narooie-Nejad, M., et al., Vitamin D receptor gene polymorphism and the risk of multiple
sclerosis in South Eastern of Iran. J Mol Neurosci, 2015. 56(3): p. 572-576.
29.
Agnello, L., et al., Vitamin D receptor polymorphisms and 25-hydroxyvitamin D in a group of
Sicilian multiple sclerosis patients. ‎Neurol. Sci, 2016. 37(2): p. 261-267.
30.
Smolders, J., et al., Fok-I vitamin D receptor gene polymorphism (rs10735810) and vitamin D
metabolism in multiple sclerosis. ‎J. Neuroimmunol, 2009. 207(1): p. 117-121.
10
31.
Agliardi, C., et al., Vitamin D receptor (VDR) gene SNPs influence VDR expression and modulate
protection from multiple sclerosis in HLA-DRB1* 15-positive individuals. Brain Behav. Immun,
2011. 25(7): p. 1460-1467.
32.
García-Martín, E., et al., Vitamin D3 receptor (VDR) gene rs2228570 (Fok1) and rs731236
(Taq1) variants are not associated with the risk for multiple sclerosis: results of a new study and
a meta-analysis. PloS one, 2013. 8(6): p. e65487.
33.
Sioka, C., et al., Vitamin D receptor gene polymorphisms in multiple sclerosis patients in
northwest Greece. J Negat Results Biomed, 2011. 10(1): p. 3.
34.
Niino, M., et al., Vitamin D receptor gene polymorphism in multiple sclerosis and the association
Downloaded by [University of Florida] at 13:58 27 October 2017
with HLA class II alleles. J Neurol Sci, 2000. 177(1): p. 65-71.
35.
Shojapuor, M., et al., Vitamin D receptor gene polymorphism in patients with multiple sclerosis
in Mar kazi Province. Arak Medical University Journal, 2012. 15(4): p. 34-39.
Figure Legend
Figure 1. Genotype patterns of four selected VDR polymorphisms using PCR-RFLP. Genotypes
were determined from lane 1-12 (left to right) as following: CC (187 bp, 55 bp), GT (242 bp, 187
bp,55 bp), TT (242 bp) for ApaI polymorphism, CC (192 bp, 105 bp), CT (297 bp, 192 bp, 105
bp), TT (207 bp) for BsmI polymorphism, TT (185 bp, 62 bp), CT (247 bp, 185 bp, 62 bp), CC
11
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(247 bp) for FokI polymorphism, and TT (699 bp), TC (699 bp, 604 bp, 95 bp), CC (604 bp, 95
bp) for TaqI polymorphism. Lane M represents 100 bp plus DNA ladder.
12
Table 1. Characteristics, including, and excluding criteria of the MS patients and control participants
MS cases
Healthy controls
Sample size
118
124
Female
82 (69.5%)
85 (68.5%)
Male
36 (30.5%)
39 (31.5%)
Age (Mean ± SD*)
37.8 ± 2.5
38.2 ± 3.6
Age range (year)
20-56
21-59
Country
Iran
Iran
City
Sanandaj, Kermanshah
Sanandaj, Kermanshah
Date of sampling
2014-2016
2015-2016
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Characteristics
Including criteria
- Lack of consanguinity between patients
- Absence of individuals with other autoimmune diseases associated with VDR gene polymorphisms such as
rheumatoid arthritis or SLE and etc. in the family
- Selection of patients based on McDonald's criteria
- All the patients were relapsing remitting.
- Have the same sex and age distribution between patients and controls
- have the same geographic area
- have the same race
Excluding criteria
- Affected related patients
- subjects with same race but different geographic area
- individuals with same geographic area but different race
- Individuals with disorders related to vitamin D deficiency such as rickets or parathyroid pathologies
- individuals who had consumed drugs or supplements containing vitamin D or calcium
* Standard deviation
13
Table 2. Distribution of genotype and allelic frequencies of the selected VDR gene polymorphisms in MS
patient and control group
Patients (118)
FokI
MAF*= 0. 0.328
Controls (124)
P value
OR* (95% CI*)
Minor allele= T
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Genotype
CC (%)
51 (43.2)
77 (62.1)
0.004
0.46 (0.28- 0.78)
CT (%)
54 (45.8)
40 (32.3)
0.032
1.77 (1.05- 2.99)
TT (%)
13 (11.0)
7 (5.6)
0.136
2.07 (0.80- 5.38)
C (%)
156 (66.1)
194 (78.2)
0.003
0.54 (0.36- 0.81)
T (%)
80 (33.9)
54 (21.8)
0.003
1.84 (1.23- 2.76)
0.818
0.555
Allele
H-W* P value
TaqI
MAF= 0.276
Minor allele=C
Genotype
TT (%)
33 (28)
62 (50)
0.0005
0.39 (0.23- 0.66)
TC (%)
61 (51.7)
49 (39.5)
0.059
1.64 (0.98- 2.73)
CC (%)
24 (20.3)
13 (10.5)
0.036
2.18 (1.05- 4.52)
T (%)
127 (53.8)
173 (69.8)
0.003
0.51 (0.39- 0.73)
C (%)
109 (46.2)
75 (30.2)
0.003
1.98 (1.36- 2.87)
0.66
0.48
Allele
H-W P value
ApaI
MAF=0.484
Minor allele=C
Genotype
CC (%)
52 (44.0)
59 (47.6)
0.584
0.87 (0.52- 1.44)
CA (%)
50 (42.4)
53 (42.7)
0.948
0.96 (0.59- 1.64)
AA (%)
16 (13.6)
12 (9.7)
0.347
1.46 (0.66- 3.24)
Allele
14
C (%)
154 (65.2)
171 (68.9)
0.394
0.85 (0.58- 1.24)
A (%)
82 (34.8)
77 (31.1)
0.394
1.18 (0.81- 1.73)
0.476
0.985
H-W P value
BsmI
MAF=0.295
Minor allele=A
Downloaded by [University of Florida] at 13:58 27 October 2017
Genotype
GG (%)
41 (34.7)
54 (43.5)
0.162
0.69 (0.41- 1.16)
GA (%)
55 (46.6)
53 (42.7)
0.552
1.17 (0.70- 1.94)
AA (%)
22 (18.7)
17 (13.8)
0.298
1.44 (0.72- 2.88)
G (%)
137 (58.1)
161 (64.9)
0.124
0.75 (0.52- 1.08)
A (%)
99 (41.9)
87 (35.1)
0.124
1.34 (0.93- 1.93)
0.641
0.493
Allele
H-W P value
* OR= Odds ratio, CI= Confidence Interval, MAF=Minor allele frequency, H-W= Hardy-Weinberg
Table 3. Analysis of combined genotypes of the selected VDR polymorphisms based on dominant
and recessive genetic models
Polymorphism
Genetic Model
OR (95% CI)
P value
FokI (rs2228570)
Dominant
0.48 (0.19- 1.26)
0.136
Recessive
2.15 (1.29- 3.60)
0.004
Dominant
0.46 (0.22- 0.95)
0.036
Recessive
2.58 (1.51- 4.40)
0.0005
Dominant
0.68 (0.31- 1.51)
0.347
Recessive
1.15 (0.69- 1.91)
0.584
Dominant
0.69 (0.35- 1.38)
0.298
Recessive
1.45 (0.86- 2.44)
0.162
TaqI (rs731236)
ApaI (rs7975232)
BsmI (rs1544410)
15
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