Sains Malaysiana 52(6)(2023): 1835-1844

http://doi.org/10.17576/jsm-2023-5206-18

Genetic Association Study of STAT4 Polymorphisms and Type 1 Diabetes in Pakistani Children

(Penyelidikan Perkaitan Genetik Polimorfisme STAT4 dan Diabetes Jenis 1 pada Kanak-kanak di Pakistan)

SANA RAFAQAT¹, JAIDA MANZOOR², MARIA ADREES¹, HAFSA HAMID¹, ASIFA KAMAL³ & RASHEEDA BASHIR^1,*

¹Department Of Biotechnology, Lahore College for Women University, 54000, Lahore, Pakistan

²Department Of Endocrinology and Diabetes, The Children's Hospital & University of Child Health Sciences, 54000, Lahore, Pakistan

³Department Of Statistics, Lahore College for Women University, 54000, Lahore, Pakistan

Diserahkan: 4 Oktober 2022/Diterima: 6 Jun 2023

Abstract

The present study investigated the relationship of STAT4 single nucleotide polymorphisms (SNPs) rs7574685, rs10181656, and rs3821236 with T1D susceptibility visiting tertiary care hospital in Lahore, Punjab, Pakistan. One hundred and fifty-five T1D patients and one hundred and five healthy individuals were enrolled. An expert endocrinologist collected the clinical data of T1D patients. The genotyping of three potential STAT4 SNPs was performed through Tetra ARMS-PCR assay. The relationship between SNPs and T1D susceptibility under several genetic models, including dominant, recessive, and codominant models, was assessed by regression analysis. All clinical features of T1D demonstrate a significant difference from control groups (P<0.01) except blindness. The characteristic biochemical analysis determined that participants with T1D had significantly higher fasting blood glucose levels and glycated hemoglobin (HbA1c) levels than the control group (P<0.01). Genetic analysis of rs7574685 depicts GT genotype was found to be the risk allele for the development of T1D when compared to the control group. For rs10181656 and rs3821236, the GC genotype and GA genotype were observed to be the risk alleles in the T1D cases as compared to the control group (P=0.04, P<0.01, respectively). Genetic models showed that the STAT4 GG genotype of rs7574685 in the dominant model (OR=1.73, 95% CI=1.05-2.86), GC genotype of rs10181656 in the codominant model (OR=2.079, 95 % CI=1.16-3.71), and AA genotype of rs3821236 showed significant risk association with T1D (OR=3.486, 95% CI=1.72-7.03). It is concluded that the risk of T1D is highly correlated with the STAT4 variants of rs7574685 and rs10181656 among children of the Pakistani population.

Keywords: Polymorphism; STAT4; T1D; variants

Abstrak

Penyelidikan ini mengkaji hubungan polimorfisme nukleotida tunggal STAT4 (SNP) rs7574685, rs10181656 dan rs3821236 dengan kerentanan T1D yang melawat hospital penjagaan tertiari di Lahore, Punjab, Pakistan. Seratus lima puluh lima pesakit T1D dan seratus lima individu yang sihat telah didaftarkan. Pakar endokrinologi mengumpul data klinikal pesakit T1D. Genotip tiga SNP STAT4 yang berpotensi dilakukan melalui ujian Tetra ARMS-PCR. Hubungan antara SNP dan kerentanan T1D di bawah beberapa model genetik, termasuk model dominan, resesif dan kodominan, telah dinilai oleh analisis regresi. Semua ciri klinikal T1D menunjukkan perbezaan yang ketara daripada kumpulan kawalan (P<0.01) kecuali buta. Analisis biokimia ciri menentukan bahawa peserta dengan T1D mempunyai paras glukosa darah puasa dan paras hemoglobin terglikasi (HbA1c) yang jauh lebih tinggi daripada kumpulan kawalan (P<0.01). Analisis genetik rs7574685 menggambarkan genotip GT didapati sebagai alel risiko untuk pembangunan T1D jika dibandingkan dengan kumpulan kawalan. Untuk rs10181656 dan rs3821236, genotip GC dan genotip GA diperhatikan sebagai alel risiko dalam kes T1D berbanding kumpulan kawalan (P = 0.04, P <0.01, masing-masing). Model genetik menunjukkan bahawa genotip STAT4 GG rs7574685 dalam model dominan (OR=1.73, 95% CI=1.05-2.86), genotip GC rs10181656 dalam model kodominan (OR=2.079, 95% CI=1.16),-3 dan genotip AA rs3821236 menunjukkan perkaitan risiko yang ketara dengan T1D (OR=3.486, 95% CI=1.72-7.03). Disimpulkan bahawa risiko T1D sangat berkorelasi dengan varian STAT4 rs7574685 dan rs10181656 dalam kalangan populasi kanak-kanak Pakistan.

Kata kunci: Polimorfisme; STAT4; T1D; varian

RUJUKAN

Abdelmajed, S.S., El-Dessouky, M.A., SalahElDin, D.S., Hassan, N.A.M., Zaki, M.E. & Ismail, S. 2021. Assessing the association of rs7574865 STAT4 gene variant and type 1 diabetes mellitus among Egyptian patients. Journal of Genetic Engineering and Biotechnology 19(1): 112.

Al-Yaarubi, S., Ullah, I., Sharef, S.W., Al Shidhani, A., Al Hanai, S., Al Kalbani, R.A. & Al Jamoodi, S. 2014. Demographic and clinical characteristics of type 1 diabetes mellitus in Omani children: Single center experience. Oman Medical Journal 100(1136): 1-4.

Batool, A., Mukhtar, M., Qayyum, I. & Shakeel, R. 2016. Association of rs2476601 and rs1544410 with onset of T1D in youngsters of Lahore, Pakistan. Journal of Bioresource Management 3(1): 8.

Baynes, H.W. 2015. Classification, pathophysiology, diagnosis and management of diabetes mellitus. J. Diabetes Metab. 6(5): 1-9.

Bi, C., Li, B., Cheng, Z., Hu, Y., Fang, Z. & Zhai, A. 2013. Association study of STAT4 polymorphisms and type 1 diabetes in Northeastern Chinese Han population. Tissue Antigens 81(3): 137-140.

Condie, A.M., Allen, T.V. & Ogle, G.D. 2020. Incidence and characteristics of childhood-and youth-onset diabetes in the Qalandarabad area in northern Pakistan. Diabetes Research and Clinical Practice 163: 108155.

Cui, G., Qin, X., Zhang, Y., Gong, Z., Ge, B. & Zang, Y.Q. 2009. Berberine differentially modulates the activities of ERK, p38 MAPK, and JNK to suppress Th17 and Th1 T cell differentiation in type 1 diabetic mice. Journal of Biological Chemistry 284(41): 28420-28429.

Cui, J., Tong, R., Xu, J., Tian, Y., Pan, J., Wang, N. & Cui, W. 2021. Association between STAT4 gene polymorphism and type 2 diabetes risk in Chinese Han population. BMC Medical Genomics 14: 1-16.

Dieude, P., Guedj, M., Wipff, J., Ruiz, B., Hachulla, E., Diot, E. & Allanore, Y. 2009. STAT4 is a genetic risk factor for systemic sclerosis having additive effects with IRF5 on disease susceptibility and related pulmonary fibrosis. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology 60(8): 2472-2479.

Farsani, S.F., Brodovicz, K., Soleymanlou, N., Marquard, J., Wissinger, E. & Maiese, B.A. 2017. Incidence and prevalence of diabetic ketoacidosis (DKA) among adults with type 1 diabetes mellitus (T1D): A systematic literature review. BMJ Open 7(7): e016587.

Goyal, R. & Jialal, I. 2020. Diabetes Mellitus Type 2. InStatPearls, Treasure Island: StatPearls Publishing. Updated 2022 June 19.

Fichna, M., Zurawek, M., Bogusz-Gorna, K., Malecki, P., Niechcial, E., Sidoruk, A. & Fichna, P. 2020. STAT4 sequence variant and elevated gene expression are associated with type 1 diabetes in Polish children. Central European Journal of Immunology 45(1): 22-28.

Gao, X., Wang, J. & Yu, Y. 2019. The association between STAT4 rs7574865 polymorphism and the susceptibility of autoimmune thyroid disease: A meta-analysis. Frontiers in Genetics 9: 708.

Glas, J., Seiderer, J., Nagy, M., Fries, C., Beigel, F., Weidinger, M. & Brand, S. 2010. Evidence for STAT4 as a common autoimmune gene: rs7574865 is associated with colonic Crohn's disease and early disease onset. PLoS ONE 5(4): e10373.

Grayson, B.L., Smith, M.E., Thomas, J.W., Wang, L., Dexheimer, P., Jeffrey, J. & Aune, T.M. 2010. Genome-wide analysis of copy number variation in type 1 diabetes. PLoS ONE 5(11): e15393.

Gauderman, W.J. 2002. Sample size requirements for matched case‐control studies of gene-environment interaction. Statistics in Medicine 21(1): 35-50.

Grimberg, J., Nawoschik, S., Belluscio, L., McKee, R., Turck, A. & Eisenberg, A. 1989. A simple and efficient non-organic procedure for the isolation of genomic DNA from blood. Nucleic Acids Research 17(20): 8390.

Hellquist, A., Sandling, J.K., Zucchelli, M., Koskenmies, S., Julkunen, H., D'Amato, M. & Kere, J. 2010. Variation in STAT4 is associated with systemic lupus erythematosus in a Finnish family cohort. Annals of the Rheumatic Diseases 69(5): 883-886.

Kiani, A.K., Jahngir, S., John, P., Bhatti, A., Zia, A., Wang, X. & Kamboh, M.I. 2015. Genetic link of type 1 diabetes susceptibility loci with rheumatoid arthritis in Pakistani patients. Immunogenetics 67: 277-282.

Kiani, A.K., John, P., Bhatti, A., Zia, A., Shahid, G., Akhtar, P. & Kamboh, M.I. 2015. Association of 32 type 1 diabetes risk loci in Pakistani patients. Diabetes Research and Clinical Practice 108(1): 137-142.

Lee, H.S., Park, H., Yang, S., Kim, D. & Park, Y. 2008. STAT4 polymorphism is associated with early‐onset type 1 diabetes, but not with late‐onset type 1 diabetes. Annals of the New York Academy of Sciences 1150(1): 93-98.

Liang, Y.L., Wu, H., Shen, X., Li, P.Q., Yang, X.Q., Liang, L. & Xie, X.D. 2012. Association of STAT4 rs7574865 polymorphism with autoimmune diseases: A meta-analysis. Molecular Biology Reports 39: 8873-8882.

Lind, M., Pivodic, A., Svensson, A.M., Ólafsdóttir, A.F., Wedel, H. & Ludvigsson, J. 2019. HbA1c level as a risk factor for retinopathy and nephropathy in children and adults with type 1 diabetes: Swedish population-based cohort study. BMJ 366: I4894.

Martinez, A., Varade, J., Marquez, A., Cenit, M.C., Espino, L., Perdigones, N. & Urcelay, E. 2008. Association of the STAT4 gene with increased susceptibility for some immune‐mediated diseases. Arthritis & Rheumatism 58(9): 2598-2602.

Mayer-Davis, E.J., Lawrence, J.M., Dabelea, D., Divers, J., Isom, S., Dolan, L. & Wagenknecht, L. 2017. Incidence trends of type 1 and type 2 diabetes among youths, 2002-2012. N. Engl. J. Med. 376: 1419-1429.

Mukhtar, M., Batool, A., Wajid, A. & Qayyum, I. 2017. Vitamin D receptor gene polymorphisms influence T1D susceptibility among Pakistanis. International Journal of Genomics 2017: 4171254.

Myrthianou, E., Zervou, M.I., Budu-Aggrey, A., Eliopoulos, E., Kardassis, D., Boumpas, D.T. & Goulielmos, G.N. 2017. Investigation of the genetic overlap between rheumatoid arthritis and psoriatic arthritis in a Greek population. Scandinavian Journal of Rheumatology 46(3): 180-186.

Orozco, G., Alizadeh, B.Z., Delgado‐Vega, A.M., González‐Gay, M.Á., Balsa, A., Pascual‐Salcedo, D. & Martín, J. 2008. Association of STAT4 with rheumatoid arthritis: A replication study in three European populations. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology 58(7): 1974-1980.

Ramírez, O.B., Rincón, J.M., Cobos, R.B., Ávila, I.A. & Bello, J.R. 2016. STAT4 confers risk for rheumatoid arthritis and systemic lupus erythematosus in Mexican patients. Immunology Letters 175: 40-43.

Sajid, N., Riaz, M., Fawwad, A. & Basit, A. 2019. Thyroid dysfunction in subjects with type 1 diabetes at a tertiary care unit of Karachi, Pakistan. Clinical Epidemiology and Global Health 7(3): 435-438.

Seddighzadeh, M., Gonzalez, A., Ding, B., Ferreiro-Iglesias, A., Gomez-Reino, J.J., Klareskog, L. & Padyukov, L. 2012. Variants within STAT genes reveal association with anticitrullinated protein antibody-negative rheumatoid arthritis in 2 European populations. The Journal of Rheumatology 39(8): 1509-1516.

Settin, A., Salama, A. & Elshazli, R. 2014. Signal transducer and activator of transcription 4 (STAT4) G> T gene polymorphism in Egyptian cases with rheumatoid arthritis. Human Immunology 75(8): 863-866.

Shera, A.S., Miyan, Z., Basit, A., Maqsood, A., Ahmadani, M.Y., Fawwad, A. & Riaz, M. 2008. Trends of type 1 diabetes in Karachi, Pakistan. Pediatric Diabetes 9(4pt2): 401-406.

Shi, Z., Zhang, Q., Chen, H., Lian, Z., Liu, J., Feng, H. & Zhou, H. 2017. STAT4 polymorphisms are associated with neuromyelitis optica spectrum disorders. NeuroMolecular Medicine 19: 493-500.

Shimura, K., Miura, J., Kawamoto, M., Kawaguchi, Y., Yamanaka, H. & Uchigata, Y. 2018. Genetic differences between type 1 diabetes with and without other autoimmune diseases. Diabetes/Metabolism Research and Reviews 34(7): e3023.

Størling, J. & Pociot, F. 2017. Type 1 diabetes candidate genes linked to pancreatic islet cell inflammation and beta-cell apoptosis. Genes 8(2): 72.

Virk, S.A., Donaghue, K.C., Cho, Y.H., Benitez-Aguirre, P., Hing, S., Pryke, A. & Craig, M.E. 2016. Association between HbA1c variability and risk of microvascular complications in adolescents with type 1 diabetes. The Journal of Clinical Endocrinology & Metabolism 101(9): 3257-3263.

Xie, Z., Chang, C., Huang, G. & Zhou, Z. 2020. The role of epigenetics in type 1 diabetes. In Epigenetics in Allergy and Autoimmunity. Advances in Experimental Medicine and Biology, edited by Chang, C. & Lu, Q. Singapore: Springer. 1253: 223-257.

Xin, Y., Yang, M., Chen, X.J., Tong, Y.J. & Zhang, L.H. 2010. Clinical features at the onset of childhood type 1 diabetes mellitus in Shenyang, China. Journal of Paediatrics and Child Health 46(4): 171-175.

Yan, N., Meng, S., Zhou, J., Xu, J., Muhali, F.S., Jiang, W. & Zhang, J. 2014. Association between STAT4 gene polymorphisms and autoimmune thyroid diseases in a Chinese population. International Journal of Molecular Sciences 15(7): 12280-12293.

Yi, J., Fang, X., Wan, Y., Wei, J. & Huang, J. 2015. STAT4 polymorphisms and diabetes risk: A meta-analysis with 18931 patients and 23833 controls. International Journal of Clinical and Experimental Medicine 8(3): 3566.

Yi, L., Wang, J.C., Guo, X.J., Gu, Y.H., Tu, W.Z., Guo, G. & Zhou, X.D. 2013. STAT4 is a genetic risk factor for systemic sclerosis in a Chinese population. International Journal of Immunopathology and Pharmacology 26(2): 473-478.

Yin, H., Borghi, M.O., Delgado‐Vega, A.M., Tincani, A., Meroni, P.L. & Alarcón‐Riquelme, M.E. 2009. Association of STAT4 and BLK, but not BANK1 or IRF5, with primary antiphospholipid syndrome. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology 60(8): 2468-2471.

Zervou, M.I., Sidiropoulos, P., Petraki, E., Vazgiourakis, V., Krasoudaki, E., Raptopoulou, A., & Goulielmos, G.N. 2008. Association of a TRAF1 and a STAT4 gene polymorphism with increased risk for rheumatoid arthritis in a genetically homogeneous population. Human Immunology 69(9): 567-571.

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