Background: The incidence of type 1 diabetes mellitus (T1DM) in Kuwait is amongst the highest in the world. Vitamin D is considered to be involved in immune modulation and its deficiency contribute to autoimmune destruction of insulin producing beta cells in T1DM patients. Vitamin D has been shown to exert its effects via a nuclear vitamin D receptor (VDR) and therefore, VDR gene may be considered a candidate for T1DM susceptibility.
Methods: The genotypes of four VDR gene polymorphisms were determined in 253 Kuwaiti Arab T1DM patients and 214 healthy controls by PCR-RFLP analysis. Serum concentrations of three autoantibodies i.e. ICA (Islet cell autoantibody), GADA (Glutamic acid decarboxylase) and INS (Insulin autoantibody) were determined by radio-immunoassays.
Results: Statistically significant differences were detected between the genotypes of two VDR gene polymorphisms (FokI, C > T, rs10735810 and TaqI, C > T, rs731236) between T1DM patients and controls (P < 0.0001). In both, the frequency of variant alleles was considerably high in T1DM than in the controls. In contrast, the VDR gene ApaI (G > T, rs7975232) and BsmI (A > G, rs1544410) polymorphisms did not show association with T1DM. The homozygous variant genotypes of FokI, ApaI and TaqI polymorphisms show significant differences between various age-of-onset subgroups while no such association was detected in the case of BsmI polymorphism. Significant differences were also noted between heterozygous genotypes of all four polymorphisms especially between 4-6y and > 6y age-of-onset subgroups of T1DM patients. Three autoantibodies, ICA (Islet cell), GADA (glutamate decarboxylase) and INS (insulin) were positively associated to, varying degrees, with T1DM in Kuwaiti Arabs harboring different VDR gene polymorphism genotypes.
Conclusions: Our results demonstrate a significant effect of two VDR gene polymorphisms (FokI and TaqI) and three autoantibodies on genetic susceptibility of T1DM in Kuwaiti Arabs along with other factors.
Keywords: Autoantibody; Genotype; Kuwait; Polymorphism; Type 1 diabetes mellitus; Vitamin D receptor gene.
Shaltout AA, Wake D, Thanaraj TA, Omar DM, Al-AbdulRazzaq D, Channanath A, et al. Incidence of type-1 diabetes has doubled in Kuwaiti children 0-14 years over the last 20 years. Ped Diabetes. 2017;18(8):761–766. doi: 10.1111/pedi.12480. - DOI - PubMed
Taha TH, Moussa MA, Rashid AR, Fenech FF. Diabetes mellitus in Kuwait. Incidence in the first 29 years of life. Diabetologia. 1983;25(4):306–308. doi: 10.1007/BF00253190. - DOI - PubMed
Karvonen M, Viik-Kajander M, Moltchanova E, Libman I, LaPorte R, Tuomilehto J. Incidence of childhood type 1 diabetes worldwide. Diabetes Care. 2000;23(10):1516–1526. doi: 10.2337/diacare.23.10.1516. - DOI - PubMed
Bener A, Al-Ali M, Hoffmann GF. High prevalence of vitamin D deficiency in young children in a highly sunny humid country: a global health problem. Minerva Pediatr. 2009;61(1):15–22. - PubMed
Gordon CM, Feldman HA, Sinclair L, Williams AL, Kleinman PK, Perez-Rossello J, Cox JE. Prevalence of vitamin D deficiency among healthy infants and toddlers. Arch Pediatr Adolesc Med. 2008;162(6):505–512. doi: 10.1001/archpedi.162.6.505. - DOI - PMC - PubMed
Jabbar Z, Aggarwal PK, Chandel N, Kohli HS, Gupta KL, Sakhuja V, Jha V. High prevalence of vitamin D deficiency in north Indian adults is exacerbated in those with chronic kidney disease. Nephrology. 2009;14(3):345–349. doi: 10.1111/j.1440-1797.2008.01082.x. - DOI - PubMed
Rovner AJ, O’Brien KO. Hypovitaminosis D among healthy children in United States: a review of the current evidence. Arch Pediatr Adolesc Med. 2008;162(6):513–519. doi: 10.1001/archpedi.162.6.513. - DOI - PubMed
Rasoul MA, Al-Mahdi M, Al-Kandari H, Dhaunsi GS, Haider MZ. Low serum vitamin- D status is associated with high prevalence and early onset of type-1 diabetes mellitus in Kuwaiti children. BMC Pediatr. 2016;16:95. doi: 10.1186/s12887-016-0629-3. - DOI - PMC - PubMed
Lemire J. 1,25-Dihydrovitamin D3-a hormone with immunomodulatory properties. Z Rheumatol. 2000;59(Suppl 1):24–27. doi: 10.1007/s003930070034. - DOI - PubMed
Mathieu C, van Etten E, Decallonne B, Guilietti A, Gysemans C, Bouillon R, Overbergh L. Vitamin D and 1,25-dihydrovitamin D3 as modulators in the immune system. J Steroid Biochem Mol Biol. 2004;89–90(1–5):449–452. doi: 10.1016/j.jsbmb.2004.03.014. - DOI - PubMed
Littorin B, Blom P, Scholin A, Arngvist HJ, Blohme G, Ekbom-Schnell A, et al. Lower levels of plasma 25-hydroxyvitamin D among young adults at diagnosis of autoimmune type 1 diabetes compared with control subjects: results from the nationwide diabetes incidence study in Sweden (DISS) Diabetologia. 2006;49(12):2847–2852. doi: 10.1007/s00125-006-0426-x. - DOI - PubMed
Pozzilli P, Manfrini S, Crino A, Picardi A, Leomanni C, Cherubini V, et al. Low levels of 25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 in patients with newly diagnosed type 1 diabetes. Horm Metab Res. 2005;37(11):680–683. doi: 10.1055/s-2005-870578. - DOI - PubMed
Nagpal S, Na S, Rathnachalam R. Noncalcemic actions of vitamin D receptor ligands. Endocr Rev. 2005;26:662–687. doi: 10.1210/er.2004-0002. - DOI - PubMed
Mathieu C, Waer M, Laureys J, Rutgeerts O, Bouillon R. Prevention of autoimmune diabetes in NOD mice by 1,25 dihydrovitamin D3. Diabetologia. 1994;37:552–558. doi: 10.1007/BF00403372. - DOI - PubMed
The EURODIAB Sub-study 2 study group. Vitamin D supplement in early childhood and risk for type 1 (insulin-dependent) diabetes mellitus. Diabetologia. 1999;42:51–54. doi: 10.1007/s001250051112. - DOI - PubMed
Hypponen E, Laara E, Reunanen A, Jarvelin MR, Virtanen SM. Intake of vitamin D and risk of type 1 diabetes: a birth-cohort study. Lancet. 2001;358:1500–1503. doi: 10.1016/S0140-6736(01)06580-1. - DOI - PubMed
Fronczak CM, Baron AE, Chase HP, et al. In utero dietary exposures and risk of islet autoimmunity in children. Diabetes Care. 2003;26:3237–3242. doi: 10.2337/diacare.26.12.3237. - DOI - PubMed
Lemos MC, Fagulha A, Coutinho E, Gomes L, Bastos M, Barros L, et al. Lack of association of vitamin D receptor gene polymorphisms with susceptibility to diabetes mellitus in the Portuguese population. Hum Immunol. 2008;69:134–138. doi: 10.1016/j.humimm.2008.01.008. - DOI - PubMed
Crofts LA, Hancock MS, Morrison NA, Eisman JA. Multiple promoters direct the tissue-specific expression of novel N-terminal variant human vitamin D receptor transcripts. Proc Natl Acad Sci U S A. 1998;95:10529–10534. doi: 10.1073/pnas.95.18.10529. - DOI - PMC - PubMed
Walters MR. Newly identified actions of the vitamin D endocrine system. Endocr Rev. 1992;13:719–764. - PubMed
Van Etten E, Verlinden L, Giulietti A, Ramos-Lopez E, Branisteanu DD, Ferreira GB, et al. The vitamin D receptor gene FokI polymorphism: functional impact on the immune system. Eur J Immunol. 2007;37:395–405. doi: 10.1002/eji.200636043. - DOI - PubMed
McDermott MF, Ramachandran A, Ogunkolade BW, Aganna E, Curtis D, Boucher BJ, et al. Allelic variation in the vitamin D receptor influences susceptibility to IDDM in Indian Asians. Diabetologia. 1997;40:971–975. doi: 10.1007/s001250050776. - DOI - PubMed
Ban Y, Taniyama M, Yanagawa T, Yamada S, Maruyama T, Kasuga A, Ban Y. Vitamin D receptor initiation codon polymorphism influences genetic susceptibility to type 1 diabetes mellitus in the Japanese population. BMC Med Genet. 2001;2:7. doi: 10.1186/1471-2350-2-7. - DOI - PMC - PubMed
Mohammadnejad Z, Ghanbari M, Ganjali R, Afshari JT, Heydarpour M, Taghavi SM, et al. Association between vitamin D receptor gene polymorphisms and type 1 diabetes mellitus in Iranian population. Mol Biol Rep. 2012;39:831–837. doi: 10.1007/s11033-011-0805-3. - DOI - PubMed
Turpeinen H, Hermann R, Vaara S, Laine AP, Simell O, Knip M, et al. Vitamin D receptor polymorphisms: no association with type 1 diabetes mellitus in the Finnish population. Eur J Endocrinol. 2003;149:591–596. doi: 10.1530/eje.0.1490591. - DOI - PubMed
Boraska V, Skrabic V, Zeggini E, Groves CJ, Buljubasic M, Peruzovic M, Zemunik T. Family-based analysis of vitamin D receptor gene polymorphisms and type 1 diabetes in population of South Croatia. J Hum Genet. 2008;53:210–214. doi: 10.1007/s10038-007-0234-2. - DOI - PubMed
Angel B, Santos JL, Carrasco E, Albala C, Perez-Bravo F. Vitamin D receptor polymorphism and susceptibility to diabetes in Chilean subjects: a case-parent study. Eur J Epidemiol. 2004;19:1085–1087. doi: 10.1007/s10654-004-1026-z. - DOI - PubMed
Khalid KE. Vitamin D receptor gene polymorphisms in Sudanese children with type 1 diabetes. AIMS Genet. 2016;3(3):167–176. doi: 10.3934/genet.2016.3.167. - DOI
Mukhtar M, Batool A, Wajid A, Qayyum I. Vitamin D receptor gene polymorphisms influence T1D susceptibility among Pakistanis. Int J Genomics. 2017. 10.1155/2017/4171254. - PMC - PubMed
Xiao XH, Liu ZL, Wang H, Sun Q, Li WH, Yang GH, Liu QY. Effects of vitamin D receptor gene polymorphisms on susceptibility to type 1 diabetes mellitus. Chinese Med Sci J. 2006;21:95–98. - PubMed
Chang TJ, LeiHH YJI, Chiu KC, Lee KC, Chen MC, Tai TY, Chuang LM. Vitamin D receptor gene polymorphisms influence susceptibility to type 1 diabetes mellitus in the Taiwanese population. Clin Endrocrinol. 2000;52:575–580. doi: 10.1046/j.1365-2265.2000.00985.x. - DOI - PubMed
Iyar A, Lanham-New S, Khoja S, Al-Ghamdi M, Al Doghaither H. Relationship between vitamin D receptor gene polymorphisms and type 1 diabetes mellitus in Saudi patients. Int J Pharmacol. 2017. 10.3923/ijp.2017.1092.1097.
Cheon C-K, Nam H-K, Lee K-H, Kim SY, Song JS, Kim C. Vitamin D receptor gene polymorphisms and type 1 diabetes mellitus in a Korean population. Pediatr Int. 2015;57:870–874. doi: 10.1111/ped.12634. - DOI - PubMed
Marti G, Audi L, Esteban C, Oyarzabal M, Chueca M, Gussinye M, et al. Association of vitamin D receptor gene polymorphism with type 1 diabetes mellitus in two Spanish populations. Medicina Clinica. 2004;123:286–289. doi: 10.1016/S0025-7753(04)74494-2. - DOI - PubMed
Thorsen SU, Mortensen HB, Carstensen B, Fenger M, Thuesen BH, Husemoen L, et al. No association between type 1 diabetes and genetic variation in the vitamin D metabolism genes: a Danish study. Pediatr Diabetes. 2014;15:416–421. doi: 10.1111/pedi.12105. - DOI - PubMed
Penna-Martinez M, Badenhoop K. Inherited variation in vitamin D genes and type 1 diabetes predisposition. Genes. 2017;8:125. doi: 10.3390/genes8040125. - DOI - PMC - PubMed
Sahin OA, Goksen D, Ozpinar A, Serdar M, Onay H. Association of vitamin D receptor polymorphisms and type 1 diabetes susceptibility in children: a meta-analysis. Endocrine Connections. 2017;6:159–171. doi: 10.1530/EC-16-0110. - DOI - PMC - PubMed
Wang G, Zhang Q, Xu N, Xu K, Wang J, He W, Yang T. Association between two polymorphisms (FokI and BsmI) of the vitamin D receptor gene and type 1 diabetes mellitus in Asian population: a meta-analysis. PLoS One. 2014;9(3):e89325. doi: 10.1371/journal.pone.0089325. - DOI - PMC - PubMed
Haider MZ, Rasoul MA, Al-Mahdi M, Al-Kandari H, Dhaunsi GS. Association of protein tyrosine phosphatase non-receptor type 22 gene functional variant C1858T, HLA-DQ/DR genotypes and autoantibodies with type-1 diabetes mellitus in Kuwaiti Arabs. PLoS One. 2018;13(6):e0198652 10.1371/journal.pone.0198652. - PMC - PubMed
Craig ME, Jefferies C, Dabelea D, Balde N, Seth A, Donaghue KC. Definition, epidemiology, and classification of diabetes in children and adolescents. Pediatr Diabetes. 2014;15(Suppl 20):4–17. doi: 10.1111/pedi.12186. - DOI - PubMed
Rojnic Putarek N, Ille J, Spehar Uroic A, Skrabic V, Stipancic G, et al. Incidence of type 1 diabetes mellitus in 0 to 14-y-old children in Croatia – 2004 to 2012 study. Pediatr Diabetes. 2015;16:448–453. doi: 10.1111/pedi.12197. - DOI - PubMed
Maniatis T. Molecular Cloning – A Laboratory Manual. In: Sambrook J, Fritsch EF, editors. Cold Spring Harbor laboratory. 2. New York: USA; 1989.
Panierakis C, Goulielmos G, Mamoulakis D, Petraki E, Papavasiliou E, Galanakis E. Vitamin D receptor gene polymorphisms and susceptibility to type 1 diabetes in Crete. Greece Clin Immunol. 2009;133:276–281. doi: 10.1016/j.clim.2009.08.004. - DOI - PubMed
Yavuz DG, Keskin L, Kiyici S, Sert M, Yazici D, Sahin I, et al. Vitamin D receptor gene BsmI, FokI, ApaI, TaqI polymorphisms and bone mineral density in a group of Turkish type 1 diabetes patients. Acta Diabetol. 2011;48:329–336. doi: 10.1007/s00592-011-0284-y. - DOI - PubMed
Dilmec F, Uzner E, Akkafa F, Kise E, van Kuilenburg ABP. Detection of VDR gene ApaI and TaqI polymorphisms in patients with type 2 diabetes mellitus using PCR-RFLP method in a Turkish population. J Diabetes & Complications. 2010;24:186–191. doi: 10.1016/j.jdiacomp.2008.12.002. - DOI - PubMed
Jurutka PW, Remus GK, Whitfield PD, Thompson JC, Hsieh H, Zitzer P, 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;14:401–420. doi: 10.1210/mend.14.3.0435. - DOI - PubMed
Moran-Auth Y, Penna-Martinez M, Badenhoop K. VDR FokI polymorphism is associated with a reduced T-helper cell population under vitamin D stimulation in type 1 diabetes patients. J Steroid Biochem Mol Biol. 2015;148:184–186. doi: 10.1016/j.jsbmb.2015.01.003. - DOI - PubMed
Uitterlinden AG, Fang Y, Van Meurs JB, Pols HA, Van Leeuwen JP. Genetics and biology of vitamin D receptor polymorphisms. Gene. 2004;338:143–156. doi: 10.1016/j.gene.2004.05.014. - DOI - PubMed
Gross C, Eccleshall TR, Malloy PJ, Villa ML, Marcus R, Feldman D. The presence of a polymorphism at the translation initiation site of the vitamin D receptor gene is associated with low bone mineral density in postmenopausal Mexican-American women. J Bone Miner Res. 1996;11:1850–1855. doi: 10.1002/jbmr.5650111204. - DOI - PubMed
Haussler MR, Whitfield GK, Haussler CA, Hsieh JC, Thompson PD, Selznick SH, et al. The nuclear vitamin D receptor: biological and molecular regulatory properties revealed. J Bone Miner Res. 1998;13:325–349. doi: 10.1359/jbmr.1998.13.3.325. - DOI - PubMed
Zemunik T, Skrabic V, Boraska V, Diklic D, Terzic IM, Capkun V, et al. FokI polymorphism, vitamin D receptor, and Interleukin-1 receptor haplotypes are associated with type 1 diabetes in the Dalmatian population. J Mol Diagnostics. 2005;7:600–604. doi: 10.1016/S1525-1578(10)60593-4. - DOI - PMC - PubMed
Mory DB, Rocco ER, Miranda WL, Kasamatsu T, Crispim F, Dib SA. Prevalence of vitamin D receptor gene polymorphisms FokI and BsmI in Brazilian individuals with type 1 diabetes and their relation to beta-cell autoimmunity and to remaining beta-cell function. Hum Immunol. 2009;70:447–451. doi: 10.1016/j.humimm.2009.03.007. - DOI - PubMed
Abd-Allah SH, Pasha HF, Hagrass HA, Alghobashy AA. Vitamin D status and vitamin D receptor gene polymorphisms and susceptibility to type 1 diabetes in Egyptian children. Gene. 2014;536:430–434. doi: 10.1016/j.gene.2013.12.032. - DOI - PubMed
Fassbender WJ, Goertz B, Weismuller K, Steinhauser B, Stracke H, Auch D, et al. VDR gene polymorphisms are overexpressed in German patients with type 1 diabetes compared to healthy controls without effect on biochemical parameters of bone metabolism. Horm Metab Res. 2002;34:330–337. doi: 10.1055/s-2002-33262. - DOI - PubMed
Pani MA, Knapp M, Donner H, Braun J, Baur MP, Usadel KH. Vitamin D receptor allele combinations influence genetic susceptibility to type 1 diabetes in Germans. Diabetes. 2000;49:504–507. doi: 10.2337/diabetes.49.3.504. - DOI - PubMed
Ogunkolade BW, Boucher BJ, Prahl JM, Bustin SA, Burrin JM, Noonan K, et al. Vitamin D receptor (VDR) mRNA and VDR protein levels in relation to vitamin D status, insulin secretory capacity, and VDR genotype in Bangladeshi Asians. Diabetes. 2002;51:2294–2300. doi: 10.2337/diabetes.51.7.2294. - DOI - PubMed
Valdivielso JM, Fernandez E. Vitamin D receptor polymorphisms and disease. Clin Chim Acta. 2006;371:1–12. doi: 10.1016/j.cca.2006.02.016. - DOI - PubMed
Silva JD, Guimaraes RL, Brandao LAV, Arauja J, Segat L, Crovella S, et al. Vitamin D receptor (VDR) gene polymorphisms and age onset in type 1 diabetes mellitus. Autoimmunity. 2013;46(6):382–387. doi: 10.3109/08916934.2013.795952. - DOI - PubMed
Motohashi Y, Yamada S, Yanagawa T, Maruyama T, Suzuki R, Niino M, et al. Vitamin D receptor gene polymorphism affects onset pattern of type 1 diabetes. J Clin Endocrinol Metab. 2003;88(7):3137–3140. doi: 10.1210/jc.2002-021881. - DOI - PubMed
Singh S, Singh UG, Agarwal NK, Singh RG, Kumar SB. Prevalence of autoantibodies and HLA DR, DQ in type 1 diabetes mellitus. J Clin Diagnostic Res. 2016;10(7):EC9–EC13. doi: 10.7860/JCDR/2016/18657.8163. - DOI - PMC - PubMed
Bingley PJ. Clinical applications of diabetes in antibody testing. J Clin Endocrinol Metab. 2010;95:25–33. doi: 10.1210/jc.2009-1365. - DOI - PubMed
Ziegler AG, Nepom GT. Prediction and pathogenesis in type 1 diabetes. Immunity. 2010;32:468–478. doi: 10.1016/j.immuni.2010.03.018. - DOI - PMC - PubMed
Wenzlau JM, Juhl K, Yu L, Moua O, Sarkar SA, Gottlieb P, et al. The cation efflux transporter ZnT8 (Slc30A8) is a major autoantigen in human type 1 diabetes. Proc Natl Acad Sci U S A. 2007;104:17040–17045. doi: 10.1073/pnas.0705894104. - DOI - PMC - PubMed
Verge CF, Stenger D, Bonifacio E, Colman PG, Pilcher C, Bingley PG, et al. Combined use of autoantibodies (IA-2ab), (GADAb, IAA, ICA) in type 1 diabetes: combinatorial islet autoantibody workshop. Diabetes. 1998;47:1857–1866. doi: 10.2337/diabetes.47.12.1857. - DOI - PubMed
Manan H, Angham AM, Sitebanat A. Genetic and auto-antibody markers in Saudi children with type 1 diabetes. Hum Immunol. 2010;71:1238–1242. doi: 10.1016/j.humimm.2010.09.008. - DOI - PubMed
Chan JCN, Yeung VTF, Chow CC, Ko GTC, Mackey IR, Rowley MJ, et al. Pancreatic beta cell function and antibodies to glutamic acid decarboxylase (anti-GAD) in Chinese patients with clinical diagnosis of insulin-dependent diabetes mellitus. Diabetes Res Clin Pract. s1996;32:27–34. - PubMed