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 Table of Contents  
Year : 2014  |  Volume : 2  |  Issue : 2  |  Page : 81-85

Association of NOD2/CARD15, DLG5, OCTN1 and toll-like receptor 4 gene polymorphisms with inflammatory bowel disease: A university hospital experience

1 Department of Internal Medicine, King Fahd Hospital of the University, Dammam, Saudi Arabia
2 Department of Internal Medicine, Prince Mohammed Center for Research and Consultation Studies, Dammam, Saudi Arabia
3 Department of Hematopathology, University of Dammam, Dammam, Saudi Arabia

Date of Web Publication18-Jul-2014

Correspondence Address:
Raed M Al-Sulaiman
Department of Internal Medicine, King Fahd Hospital of the University, University of Dammam, P.O. Box 4225, Dammam 31491
Saudi Arabia
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/1658-631X.136988

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Background: Both genetic and environmental factors play major roles in the development of inflammatory bowel disease (IBD). Recent studies have identified a number of genetic susceptibility loci for Crohn's disease (CD) and ulcerative colitis (UC).
Objectives: The present study aimed at examining the association of nine polymorphisms in four different genes with the development of CD and UC in a sample of Saudi patients with IBD.
Materials and Methods: All gene polymorphisms were identified by polymerase chain reaction and by direct sequencing. Allele and genotype frequencies of polymorphisms of NOD2/CARD15 (R702W, G908R, L1007finsC), Toll-like receptor 4 (TLR4) (D299G, T399I), OCTN promoter (C1672, G207C) and DLG5 (G113A, C4136A) genes were determined in Saudi subjects with CD (51), UC (26) and in 75 normal controls.
Results: Out of the nine polymorphisms studied in four loci, only two polymorphisms in two different loci were found to have increased in patients compared with the control subjects. The CT genotype of TLR4 T3991 was over represented in patients with CD or UC compared to that in controls (odds ratios [OR], 5.63:95% confident interval [CI], 1.19-26.69; P = 0.03). In addition, the GA genotype of DLG5 G113A was over represented in patients with CD or UC compared with that in controls (OR, 4.72:95% CI 2.30-9.66; P = 0.0001). However, there were no significant associations found between all other polymorphisms studied and the susceptibility of CD or UC found in the Saudi population.
Conclusion: Our finding indicates that association of IBD with nine gene polymorphisms was only significant in two of these polymorphic variants.

  Abstract in Arabic 

ملخص البحث:
تلعب العوامل الجينية والبيئية دورًا مهما في تكون أمراض الأمعاء التقرحية، وقد بينت الدراسات الحديثة عددًا من المواقع ذات القابلية الوراثية لمرض كرون والقولون التقرحي وقد هدفت هذه الدراسة المستقبلية والتي أجريت في مستشفى الملك فهد الجامعي بالخبر وشملت 77 مريضًا لبحث ارتباط تعدد الأشكال في أربعة جينات مختلفة بتطور مرض كرون والقولون التقرحي لدى عدد من المرضى السعوديين الذين يعانون من التهابات الأمعاء التقرحية.
لم توضح الدراسة وجود ارتباط هام بين كل المواضع الوراثية المدروسة وقابلية حدوث التهابات الأمعاء التقرحية لدى العينة المدروسة. وتشير الدراسة أن ارتباط أمراض الأمعاء التقرحية مع التسعة الجينات متعددة الأشكال كان مهما فقط لدى اثنين من المتغيرات متعددة الأشكال.

Keywords: Genetic susceptibility, inflammatory bowel disease, Saudi Arabia

How to cite this article:
Al-Sulaiman RM, Ismail MH, Yasawy MI, Al-Ateeq SA, Abdelrashid MM, Hussameddin AM, Al-Nafie AM, Al-Ali AK, Akhtar MS, Al-Quorain AA. Association of NOD2/CARD15, DLG5, OCTN1 and toll-like receptor 4 gene polymorphisms with inflammatory bowel disease: A university hospital experience. Saudi J Med Med Sci 2014;2:81-5

How to cite this URL:
Al-Sulaiman RM, Ismail MH, Yasawy MI, Al-Ateeq SA, Abdelrashid MM, Hussameddin AM, Al-Nafie AM, Al-Ali AK, Akhtar MS, Al-Quorain AA. Association of NOD2/CARD15, DLG5, OCTN1 and toll-like receptor 4 gene polymorphisms with inflammatory bowel disease: A university hospital experience. Saudi J Med Med Sci [serial online] 2014 [cited 2022 Dec 7];2:81-5. Available from: https://www.sjmms.net/text.asp?2014/2/2/81/136988

  Introduction Top

The inflammatory bowel disease (IBD) is characterized by acute and chronic inflammatory changes in either the small or large bowel, or in both. It has been shown that both genetic and environmental factors play a major role in the development of IBD, including ulcerative colitis (UC) and Crohn's disease (CD). The prevalence of IBD is on the increase worldwide with an estimated prevalence in the western world that is twice as high as in Eastern Europe. [1]

The etiology of the disease is unclear, but a number of studies have shown that genetic factors play a central role in the development of IBD. However, genetic drift cannot account for the increase in the incidence of IBD in various populations. Hence, other factors including environmental, immunological and microbes have been implicated in the precipitation of the disease. [2],[3] Genome wide association studies have identified a number of genetic susceptibility loci for CD and UC. Polymorphisms in four genes were identified to be associated with susceptibility to CD and UC. Three NOD2/CARD15 polymorphisms (R702W, G908R and L1007fsinsC), two OCTN1/SLC22A5 polymorphisms (C16272T and G207C), two toll-like receptor 4 (TLR4) polymorphisms (D299G and T399I) and two DLG5 polymorphisms (G113A and C4136A) have been identified as susceptibility genes for IBD in various populations. [4],[5],[6] However, other studies have shown that these polymorphisms are not associated with development of IBD in other populations. [7],[8],[9],[10],[11],[12]

The incidence of IBD is also on the increase in Saudi Arabia. It has been shown that although the incidence of UC is at a steady state, the incidence of CD is increasing drastically. Until date, there is no report of whether these specific polymorphisms are associated with the incidence of IBD in this population. [13],[14],[15],[16],[17],[18],[19],[20] In addition, the reduction in the incidence of IBD necessitates improved risk stratification and new strategies for prevention of the disease. The aim of the present study is to determine the prevalence of these polymorphic sites in normal Saudi individuals and in patients with IBD.

  Materials and Methods Top

A prospective case-control study was conducted in 77 patients with confirmed IBD. The diagnosis was confirmed by endoscopic and histopathological studies (51 with CD and 26 with UC) residing in the Eastern Province of Saudi Arabia between 2010 and 2012. Normal age- and gender-matched subjects (75) were selected at random from the general population. The sole exclusion criteria for control subjects were past or family history of IBD disease. Informed written consents were obtained from both the patient and the control groups prior to participation in the study, which was approved by the Ethics Committee of the Research Center. Blood samples were collected in ethylenediaminetetraacetic acid tubes and were frozen until analysis. Genomic deoxyribonucleic acid was obtained from 300 ul whole blood using QIAamp Blood Kit (Qiagen, Hilden, Germany) according to the manufacturer's protocol.

Genotype determination was accomplished using restriction fragment length polymorphism procedures as previously described. [21],[22],[23] Briefly, the specific region of each loci spanning the polymorphic sites were amplified. Primer pairs (forward and reverse) for each locus are shown in [Table 1].
Table 1: Primers and restriction enzymes used to determine each SNP

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Genotype and allele frequencies were estimated by gene counting and expressed as percentages of the total. The χ2 test was used to compare differences between the groups. A difference was considered to be statistically significant when P < 0.05. To study association and adjust for confounders, a logistic regression analysis was carried out. Odds ratios (OR) are given with 95% confidence interval (CI) of MI in relation to β2-AR polymorphic genotypes.

  Results Top

A total of 152 subjects (51 patients with CD; 26 patients with UC and 75 normal healthy controls) were included in the study. The baseline characteristics of the study groups are shown in [Table 2].
Table 2: Characteristics of subjects included in the

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The obtained data showed no difference in the prevalence of different polymorphisms between CD group and UC group. Therefore, the results of the two groups were pooled together. There was a lack of association in seven of the polymorphisms studied with CD and UC. These polymorphisms included NOD2/CARD15 (R702W, G908R, L1007finsC), TLR4 (D299G), OCTN promoter (C1672, G207C) and DLG5 (C4136A) genes (P > 0.05 for all polymorphisms).

However, the remaining two polymorphisms were found to be increased in patients compared with the control subjects [Table 3]. The CT genotype of TLR4 T3991 was over-represented in patients with CD and UC compared to that in controls (OR, 5.63:95% CI: 1.19-26.69; P = 0.03), whereas he GA genotype of DLG5 G113A was over represented in patients with CD or UC compared to that in controls (OR, 4.72:95% CI: 2.30-9.66; P = 0.0001).
Table 3: Genotype frequency of various polymorphisms in patients with IBD and in normal controls

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  Discussion Top

Although CD and UC were considered to be mainly prevalent in the Western world, recent studies have shown that these diseases are also prevalent in other populations, including Saudi Arabia and the surrounding regions. [13],[14],[15],[16],[17],[18],[19],[20],[24],[25],[26] Moreover, epidemiological studies have shown that the incidence of heterogeneous diseases, including CD and UC is on the increase worldwide, including Saudi Arabia. [1],[13],[19] These studies also showed that the rate of increase in CD exceeds the increase in UC. This is in line with reports from other parts of Asia which also indicate the incidence of IBD is on the increase in these countries. This increase has been attributed to recent changes in lifestyle, dietary habits and environmental factors. Hence, implementation of steps toward reducing the incidence of CD and UC and improving the treatment of IBD necessitates improved risk stratification and new strategies.

With the conclusion of the human genome project, new insights have emerged on how genes influence physiological functions involved in modulation of specific phenotypes. It has been shown that genetic polymorphisms may play a vital role in the susceptibility or severity of diseases. Genome wide association studies and other genetic investigations have shown that there is increasing evidence for the implication of genetic factors in the development of IBD. [27],[28],[29],[30],[31],[32],[33] These studies have now identified over 100 confirmed IBD susceptibility loci in Western populations. [29] However, not all these single nucleotide polymorphisms were found to be associated with IBD in other populations. [7],[8],[9],[10],[29],[30]

Polymorphisms in four loci were identified to be associated with susceptibility to CD and UC. Three NOD2/CARD15 polymorphisms (R702W, G908R and L1007fsinsC) were found to be associated with IBD in the Caucasian populations. [5],[6] However, these same polymorphisms were not found to be associated with CD in Japanese and Chinese populations. [7],[8] Our results are in agreement with the latter studies in that we did not find any association between these polymorphisms and CD or UC in Saudi patients. Similarly, two OCTN1/SLC22A5 polymorphisms (C16272T and G207C) which have been found to be associated with CD or UC in Caucasian populations were not found to be associated with CD or UC in the present population. [31],[32] In addition, the TLR4 polymorphisms (D299G) and DLG5 polymorphisms (C4136A) were also found not to be associated with CD and UC in Saudi patients. However, the TLR4 polymorphisms (T399I) and DLG5 polymorphisms (G113A) were found to be increased in Saudi patients with CD and UC. This is in agreement with the majority of studies conducted on Caucasian populations but conflicts with the results of studies conducted on Italian and Asian populations. [33],[34],[35] similar study from the Central province of KSA, which have found association with CARD15/NOD2 variant alleles and the CD14-159C/T polymorphism and in contrast no evidence that the TLR4 (Thr399Il) can be considered risk factors for CD. [36]

These conflicting conclusions maybe due to different influencing factors, such as study design and ethnicity of the study populations. The presented data suggest a need for further work and larger studies to determine other genetic and environmental factors influencing CD susceptibility and behavior in the Saudi population. It is still early to rely on those genetic factors for diagnosing IBD but it can indicate individual at risk.

  References Top

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  [Table 1], [Table 2], [Table 3]

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