This study was conducted to explore whether VDR gene polymorphisms are associated with psoriasis susceptibility and clinical response to calcipotriol in psoriatic patients. The targeted DNA fragment was amplified for Polymerase Chain Reaction (PCR) in a 25µl reaction mixture by using following primers: for rs10735810 polymorphic site “Forward primer (F.P): AGCTGGCCCTGGCACTGACTCTGGCTCT and Reverse Primer (R.P): ATGGAAACACCTTGCTTCTTCTCCCTC”, for rs7975232 and rs731236 polymorphic site “F.P: CAGAGCATGGACAGGGAGCAA and R.P: GCAACTCCTCATGGCTGAGGTCTC”, and for rs1544410 polymorphic site “F.P: CAACCAAGACTACAAGTACCGCGTCAGTGA and R.P: AACCAGCGGGAAGAGGTCAAGGG.” For all primers, the PCR reaction included 35 cycles, which consist of initial denaturation at 95°C for 5:00 mins, denaturation at 95°C for 45s, annealing at 68°C (rs10735810), 65°C (rs7975232 and rs731236), and 55.9°C(rs1544410) for 45s, and extension at 72°C for 30s followed by final denaturation at 72°C for 10 mins. Vitamin D deficiency has been associated with increased incidence of diabetes, both in humans and in animal models. Our study has shown lower mean levels of 25(OH)D in patients with T2DM (28.54 ± 10.02) in comparison with control subjects (44.65 ± 7.19), p < 0.001. VDR gene polymorphism leads to dysfunctioning of 1, 25(OH)2D3ultimately disease onset. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Nucleotide change on rs10735810 site leads to change of tryptophan with arginine. Wiad Lek, 2020. It acts as an immunoregulator by activating human lymphocytes that express its steroid receptor, vitamin D receptor (VDR), and inhibit antibody production and interleukin-2 along with suppression of cytotoxic lymphocytes and their proliferation [2]. In Pakistan, 70% of marriages are cousin marriage which is the major reason of transferring genetic disorders to next generation. Blood samples (3cc) from each case and control were collected in EDTA-coated tubes and DNA was extracted by the modified organic extraction method [7]. Vitamin D receptor gene BsmI B allele may be a risk factor for systemic lupus erythematosus (SLE) onset among overall populations and Asians, and FokI FF genotype is a risk factor for SLE susceptibility in Asians. There is lack of baseline data that is required with reference to our population. By continuing you agree to the use of cookies. Vitamin D insufficiency was more prevalent in T2DM 58% than in healthy control subjects 4%. The current study was ethically approved by the Punjab University Advanced Studies and Research Board, Lahore, Pakistan. FBG, 2 h –PPG, fasting lipids, Hb A1c, calcium, phosphorus, urea, creatinine, ALT, AST were measured. The tubes were incubated at 55°C for 5h (FokI), 37°C for 16h (ApaI), 65°C for 3h (TaqI), and 37°C for 5h (BsmI), followed by thermal inactivation of restriction enzymes at 80°C (FokI and TaqI) and 65°C (ApaI and BsmI), for 20 mins. 1, 25 dihydroxy vitamin D3 is pleiotropic steroid formed in skin by sunlight exposure from its precursor [1]. Purpose . The clinical characteristics of the participants were presented in Table 1. Briefly, a reaction mixture of 25 µl containing 10 µl of each PCR product was mixed with 2 µl of Tango buffer, 1 µl of restriction enzyme, and 12 µl of DEPC water. Vitamin D receptor (VDR) gene is the first reported gene potentially associated with IDD risks . It binds to cytoplasm VDR and forms complex after entering the cell. The active VDR hormone is 1,25-dihydroxycholecalciferol or vitamin D 3. Moreover, vitamin D level is not significantly inadequate but VDR gene polymorphism is a significant risk factor of RA as well as OA onset in Pakistani population. Amino acid alignment on Mega 6 software showed that change in nucleotide on rs10735180 polymorphic site ultimately leads to the change of tryptophan with arginine. Current study demonstrated the strong association of VDR gene polymorphism (rs10735810, rs7975232, rs731236, and rs1544410) with the onset of RA and OA in studied population. The functions of these vitamin D receptor polymorphisms have been comprehensively studied in T2DM patients [51]. Enters the nucleus upon vitamin D3 binding where it forms heterodimers with the retinoid X receptor/RXR (PubMed:28698609). Majority of them cause VDR to be unable to bind to 1, 25-OH-D. Over 63 polymorphisms on VDR gene were reported in different populations out of which rs10735810, rs1544410, rs7975232, and rs731236 were extensively studied [6], whereas very few studies investigate association of VDR gene polymorphism with OA. DNA was extracted from each blood sample and amplified by using gene specific primers. Nanodrop was performed for DNA quantification by the nanodrop (Thermo 2000) method. The variation on rs731236, rs1544410, and rs7975232 polymorphic site will not lead to the change of amino acid sequence. It was demonstrated that high BMI has been significantly associated with arthritis onset. We sequenced the VDR gene region … After the entrance of vitamin D2 or D3, it forms complex with the vitamin D binding protein (VDBP) and then makes its way to the liver. Authors are thankful to Higher Education Commission and worthy Vice Chancellor, University of the Punjab, and Lahore, for providing funding for accomplishment of research. Glover et al. The VDR gene provides instructions for making a protein called vitamin D receptor (VDR), which allows the body to respond to vitamin D. This vitamin can be acquired from foods in the diet or made in the body with help from sunlight exposure. As for the association of vitamin D receptor (VDR) gene polymorphisms with susceptibility to pediatric asthma, results of published studies yielded conflicts. Hughes et al. Linkage disequilibrium and haplotype were calculated to study their association with RA and OA by SHEsis (http://analysis.bio-x.cn/myAnalysis.php). Both positive paternal and maternal family history of arthritis were significant risk factors of disease development. The inclusion criteria for RA patients include patients with positive RF factor and who are eligible for Rituximab therapy according to UK NICE guidelines; 2010 ACR/EULAR Rheumatoid Arthritis classification criteria for a diagnosis of RA; participants of 18 years of age or over must be capable of giving informed consent. Latacz M, et al. On the other hand, in OA subjects, haplotype CGAT, CGAT, CGGT, CTGA, TGAT, TGGA, TTAA, and TTGA were associated with disease onset. Serum was separated and vitamin D level as determined from each sample by ELISA. High BMI also influences OA severity as in comparison to normal or underweight subjects, obese people have significant knee joint degradation [15]. VDR genotyping for rs10735810, rs1544410, rs7975232, and rs731236 was performed by direct sequencing method and Restriction Fragment Length Polymorphism (PCR-RFLP). As a result of ELISA, it was found that in serum 25(OH)2D3 was sufficient among RA, OA, and controls (30 ng/ml – 100ng/ml) and there was no significant difference in 25(OH)2D3 level among the studied groups (Figure 1). rs7975232 along with rs1544410 (D’=0.629, r2=0.103) and rs731236 along with rs1544410 (D’=0.995, r2=0.618) were also associated with OA onset. The purpose of current study was to evaluate the effect of vitamin D level and VDR gene polymorphism on rheumatoid arthritis and osteoarthritis. J. Sambrook, E. F. Fritsch, and T. Maniatis, Y. Arnson, H. Amital, and Y. Shoenfeld, “Vitamin D and autoimmunity: new aetiological and therapeutic considerations,”, Y. J. Cheng, J. M. Hootman, L. B. Murphy, G. A. Langmaid, and C. G. Helmich, “Prevalence of doctor-diagnosed arthritis and arthritis-attributable activity limitation-United States, 2007-2009,”, Centers for Disease Control and Prevention (CDC), “Prevalence of obesity among adults with arthritis---United States, 2003--2009,”, M. Pedersen, S. Jacobsen, M. Klarlund et al., “Environmental risk factors differ between rheumatoid arthritis with and without auto-antibodies against cyclic citrullinated peptides,”, A. Wesley, C. Bengtsson, A.-C. Elkan, L. Klareskog, L. Alfredsson, and S. Wedrén, “Association between body mass index and anti-citrullinated protein antibody-positive and anti-citrullinated protein antibody-negative rheumatoid arthritis: Results from a population-based case-control study,”, A. H. M. Van Der Helm-van Mil, S. M. Van Der Kooij, C. F. Allaart, R. E. M. Toes, and T. W. J. Huizinga, “A high body mass index has a protective effect on the amount of joint destruction in small joints in early rheumatoid arthritis,”, P. W. Lementowski and S. B. Zelicof, “Obesity and osteoarthritis.,”, C. Muehleman, A. Margulis, W. C. Bae, and K. Masuda, “Relationship between knee and ankle degeneration in a population of organ donors,”, H. Bliddal and R. Christensen, “The management of osteoarthritis in the obese patient: Practical considerations and guidelines for therapy,”, P. Hanvivadhanakul and J. Singhea, “The Relationship of Serum Vitamin D Level and Disease Activity in Rheumatoid Arthritis Patients,”, B. M. Baskan, F. G. Yurdakul, E. Aydn, F. Sivas, and H. Bodur, “Effect of vitamin D levels on radiographic knee osteoarthritis and functional status,”, J. Vojinovic, A. Tincani, A. Sulli et al., “European multicentre pilot survey to assess vitamin D status in rheumatoid arthritis patients and early development of a new Patient Reported Outcome questionnaire (D-PRO),”, J. Lin, J. Liu, M. L. Davies, and W. Chen, “Serum vitamin D level and rheumatoid arthritis disease activity: review and meta-analysis,”, M. I. Abd-Elazeem and R. A. Mohamed, “Neutrophil-lymphocyte and platelet-lymphocyte ratios in rheumatoid arthritis patients: Relation to disease activity,”, D. Sanghi, A. Mishra, A. C. Sharma et al., “Does vitamin D improve osteoarthritis of the knee: a randomized controlled pilot trial,”, T. L. Glover, B. R. Goodin, C. D. King et al., “A cross-sectional examination of Vitamin D, obesity, and measures of pain and function in middle-aged and older adults with knee osteoarthritis,”, C. A. Hitchon, Y. Vitamin D receptor, oestrogen receptor-alpha gene and interleukin-1 receptor antagonist gene polymorphisms in Hungarian patients with primary biliary … Current study demonstrated that no significant difference was observed in serum vitamin D level in RA, OA, and control subjects. Nurse health study also reported that risk of RA increases with obesity before 55 οf age (HR 1.65; 95% CI 1.34 to 2.05 [13]. Associations were found only between VDR FokI gene polymorphism and susceptibility to Egyptian patients with T2DM. VDR is considerably involved in the regulation of homeostatic processes. Copyright © 2021 Elsevier B.V. or its licensors or contributors. Meanwhile, contrary to this, no significant association was reported in Korean (rs1544410, rs7975232), Spanish (rs7975232, rs731236, and rs1544410), Hungarian (rs1544410), French (rs1544410, rs7975232), Tunisian (rs7975232, rs731236, and rs1544410), Turkish (rs1544410, rs7975232, and rs10735810), Egyptian (Mansoura) (rs10735810), Indian (rs10735810), German (rs10735810, rs1544410, and rs7975232), and Egyptian (Zagazig) (rs1544410) populations [25–34]. Maryam Mukhtar, Nadeem Sheikh, Saira Kainat Suqaina, Andleeb Batool, Naz Fatima, Rabia Mehmood, Sabeen Nazir, "Vitamin D Receptor Gene Polymorphism: An Important Predictor of Arthritis Development", BioMed Research International, vol. Published by Elsevier Inc. Journal of Clinical & Translational Endocrinology, https://doi.org/10.1016/j.jcte.2018.11.005. Similarly, another study reported that serum 25 (OH) D levels were not associated with the radiographic knee OA severity and its functional assessment [18]. Vitamin D hormone is needed to break down into GcMAF which is then needed to attack stealth pathogens, viruses, and cancer cells. Vitamin D is an anti-inflammatory molecule and has a role in prevention of arthritis development. The odd ratio and coefficients interval calculated were 1.42 and 0.82~2.44 in RA, whereas they were 1.11 and 0.64 ~1.89 in OA individuals. Location and map of linkage disequilibrium (LD) in SNPs at, Vitamin D Receptor Gene Polymorphism: An Important Predictor of Arthritis Development, Cell and Molecular Biology Lab, Department of Zoology, University of the Punjab, Quaid-i-Azam Campus, Lahore 54590, Pakistan, Department of Zoology, Government College University, Lahore 54000, Pakistan, Haplotype: rs10735810; rs7975232; rs1544410; rs731236, F. A. Houssiau, C. Vasconcelos, D. D'Cruz et al., “Immunosuppressive therapy in lupus nephritis: The Euro-Lupus Nephritis Trial, a randomized trial of low-dose versus high-dose intravenous cyclophosphamide,”, S. Christakos, P. Dhawan, A. Verstuyf, L. Verlinden, and G. Carmeliet, “Vitamin D: metabolism, molecular mechanism of action, and pleiotropic effects,”, M. C. Medici, F. Tummolo, A. Calderaro et al., “Identification of the novel Kawasaki 2014 GII.17 human norovirus strain in Italy, 2015,”, A. Boonstra, F. J. Barrat, C. Crain, V. L. Heath, H. F. J. Savelkoul, and A. O'Garra, “1, 25-Dihydroxyvitamin D3 has a direct effect on naive CD4+ T cells to enhance the development of Th2 cells,”, P. Lal, Z. Su, C. T. J. Holweg et al., “Inflammation and autoantibody markers identify rheumatoid arthritis patients with enhanced clinical benefit following rituximab treatment,”. BMI = Body Mass Index; n = No of individuals; RA = Rheumatoid arthritis; OA = Osteoarthritis. The minimum detectable dose (MDD) of kit used was 0.67ng/ml. Expression of the vitamin D receptor is increased in the hypertrophic heart. 25-hydroxyvitamin D (25[OH]D) is then transported to the kidney where it again undergoes hydroxylation by the action of 1-hydroxylase enzyme and forms 1,25 dihydroxy vitamin D (1,25[OH]2D) (active hormonal form) and then bounds to the VDBP to reach the target cell. Vitamin D is an anti-inflammatory molecule and has a role in prevention of arthritis development. All participants were clinically diagnosed with RA and OA by a physician according to WHO criteria. Genetic test of genetic variants in RA, OA and controls. Single-site analysis was performed and was presented in Table 2. Nutrients, 2021 Jan 11. Vitamin D receptor (VDR) gene polymorphisms may be associated with the risk of OA, however, evidence for this is controversial. Vitamin D3 is known to play a key role in calcium homeostasis that is in metabolism of minerals by bonding to its receptor (VDR). Until now four single nucleotide polymorphisms of vitamin D receptor gene ( VDR ), rs2228570 ( Fok I), rs1544410 ( Bsm I), rs7975232 ( Apa I), and rs731236 ( Taq I), have been studied in autoimmune thyroid disorders, with conflicting results. Ultimately, all these effects of vitamin D influence rheumatoid arthritis (RA) as well as osteoarthritis (OA). Vitamin D receptor polymorphisms: Four allelic variants of vitamin D receptor gene have been recognized: Apa I, Fok I, Bsm I and Taq I [72]. rs731236, rs1544410, and rs7975232 if passed together to next generation raised risk of OA development as compared to that of controls (D’=0.967, r2=0.670) followed by rs1544410, rs10735180, and rs7975232 (D’=0.650, r2=0.126). The change in amino acid sequences was determined by aligning sequences in Mega 6 software. Furthermore, vitamin D 3 analogue has anti‐psoriatic activity; however, the clinical response is variable. This complex enters in nucleus where it heterodimerizes with retinoic acid X receptor and enhances vitamin D-dependent genes transcription crucial in bone and calcium metabolism [4]. It was observed that, on rs10735810 polymorphic site on exon 2, allele ‘C’ acts as risk allele and is significantly associated with the onset of RA as well as OA (p = 0.016).