microRNA Expressions in CD4+ and CD8+ T-cell Subsets in Autoimmune Thyroid Diseases

Abstract

Graves’ disease (GD) and Hashimoto’s thyroiditis (HT) are the most common autoimmune thyroid diseases (AITD). MicroRNAs (miRNAs) critically control gene-expression and play an important role in regulating the immune response. The aim of this study was to prove significant variations of key immunoregulatory miRNAs in peripheral blood mononuclear cells (PBMCs) and in CD4+ and CD 8+ T-cells of AITD patients. Selected miRNAs were amplified by a semiquantitative SYBR Green PCR from PBMCs and purified CD4+ and CD 8+ T-cells of 59 patients with GD, HT, and healthy controls. Both GD and HT showed significantly decreased miRNA 200a_1 and miRNA 200a2* in CD4+-T-cells (mean relative expression 12,57 in HT vs. 19.40 in control group (CG), p=0.0002; 12,10 in GD vs. 19.40 in CG, p=0.0002) and in CD8+-T-cells (13.13 in HT vs. 18,12 in CG, p=0.02; 11.66 in GD vs. 18.12 in CG, p=0.0002). GD and HT showed significantly decreased miRNA 155_2 and miRNA 155*_1 in HT in CD8+-T-cells (10.69 in HT vs. 11.30 in CG, p=0.01; 10.40 in GD vs. 11.30 in CG, p=0.005). This study confirms significant variations of miRNA200a and miRNA155 in patients suffering from GD and HT in vivo in CD4+ T-cells and CD8+ T-cells. These data may help to better understand the gene regulations in the causative cells causing these autoimmune processes. They extend our very limited knowledge concerning miRNAs in thyroid diseases.

Key words

micro-RNA - autoimmune thyroid ­diseases - Graves’ disease - Hashimoto’s thyroiditis - immunology

• References

• 1 Pearce EN, Farwell AP, Braverman LE. Thyroiditis. N Engl J Med 2003; 348: 2646-2655
• 2 Hollowell JG, Staehling NW, Flanders WD et al. Serum TSH, T(4), and thyroid antibodies in the United States population (1988 to 1994): National Health and Nutrition Examination Survey (NHANES III). J Clin Endocrinol Metab 2002; 87: 489-499
• 3 Ehlers M, Thiel A, Bernecker C et al. Evidence of a combined cytotoxic thyroglobulin and thyroperoxidase epitope-specific cellular immunity in Hashimoto’s thyroiditis. J Clin Endocrinol Metab 2012; 97: 1347-1354
• 4 Ng HP, Kung AWC. Induction of autoimmune thyroiditis and hypothyroidism by immunization of immunoactive T cell epitope of thyroid peroxidase. Endocrinology 2006; 147: 3085-3092
• 5 Itoh M, Takahashi T, Sakaguchi N et al. Thymus and autoimmunity: production of CD25+CD4+ naturally anergic and suppressive T cells as a key function of the thymus in maintaining immunologic self-tolerance. J Immunol 1999; 162: 5317-5326
• 6 Hidaka Y, Amino N, Iwatani Y et al. Increase in peripheral natural killer cell activity in patients with autoimmune thyroid disease. Autoimmunity 1992; 11: 239-246
• 7 Dalan R, Leow MKS. Immune manipulation for Graves’ disease: Re-exploring an unfulfilled promise with modern translational research. Eur J Intern Med 2012;
• 8 Fountoulakis S, Tsatsoulis A. On the pathogenesis of autoimmune thyroid disease: a unifying hypothesis. Clin Endocrinol (Oxf) 2004; 60: 397-409
• 9 Matei I, Matei L.. Cytokine patterns and pathogenicity in autoimmune diseases. Rom J Intern Med 2002; 40: 27-41
• 10 Slowik M, Urbaniak-Kujda D, Bohdanowicz-Pawlak A et al. CD8+CD28-lymphocytes in peripheral blood and serum concentrations of soluble interleukin 6 receptor are increased in patients with Graves’ orbitopathy and correlate with disease activity. Endocr Res 2012; 37: 89-95
• 11 Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004; 116: 281-297
• 12 Lee RC, Feinbaum RL, Ambros V. The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 1993; 75: 843-854
• 13 He L, Hannon GJ. MicroRNAs: small RNAs with a big role in gene regulation. Nat Rev Genet 2004; 5: 522-531
• 14 Xiao C, Rajewsky K. MicroRNA control in the immune system: basic principles. Cell 2009; 136: 26-36
• 15 Croce CM, Calin GA. miRNAs, cancer, and stem cell division. Cell 2005; 122: 6-7
• 16 Li Q-J, Chau J, Ebert PJR et al. miR-181a is an intrinsic modulator of T cell sensitivity and selection. Cell 2007; 129: 147-161
• 17 Neilson JR, Zheng GXY, Burge CB et al. Dynamic regulation of miRNA expression in ordered stages of cellular development. Genes Dev 2007; 21: 578-589
• 18 Zhou B, Wang S, Mayr C et al. miR-150, a microRNA expressed in mature B and T cells, blocks early B cell development when expressed prematurely. Proc Natl Acad Sci USA 2007; 104: 7080-7085
• 19 Chen C-Z, Li L, Lodish HF et al. MicroRNAs modulate hematopoietic lineage differentiation. Science 2004; 303: 83-86
• 20 Kim VN, Han J, Siomi MC. Biogenesis of small RNAs in animals. Nat Rev Mol Cell Biol 2009; 10: 126-139
• 21 O’Carroll D, Mecklenbrauker I, Das PP et al. A Slicer-independent role for Argonaute 2 in hematopoiesis and the microRNA pathway. Genes Dev 2007; 21: 1999-2004
• 22 Harfe BD, McManus MT, Mansfield JH et al. The RNaseIII enzyme Dicer is required for morphogenesis but not patterning of the vertebrate limb. Proc Natl Acad Sci USA 2005; 102: 10898-10903
• 23 Kanellopoulou C, Muljo SA, Kung AL et al. Dicer-deficient mouse embryonic stem cells are defective in differentiation and centromeric silencing. Genes Dev 2005; 19: 489-501
• 24 Chong MMW, Rasmussen JP, Rudensky AY et al. The RNAseIII enzyme Drosha is critical in T cells for preventing lethal inflammatory disease. J Exp Med 2008; 205: 2005-2017
• 25 Koralov SB, Muljo SA, Galler GR et al. Dicer ablation affects antibody diversity and cell survival in the B lymphocyte lineage. Cell 2008; 132: 860-874
• 26 Bernecker C, Lenz L, Ostapczuk MS et al. MicroRNAs -146a1, -155 2, -200a1 are Regulated in Autoimmune Thyroid Diseases. Thyroid 2012;
• 27 Ehlers M, Thiel A, Bernecker C et al. Evidence of a combined cytotoxic thyroglobulin and thyroperoxidase epitope-specific cellular immunity in Hashimoto’s thyroiditis. J Clin Endocrinol Metab 2012; 97: 1347-1354
• 28 Taganov KD, Boldin MP, Chang K-J et al. NF-kappaB-dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. Proc Natl Acad Sci USA 2006; 103: 12481-12486
• 29 Perry MM, Moschos SA, Williams AE et al. Rapid changes in microRNA-146a expression negatively regulate the IL-1beta-induced inflammatory response in human lung alveolar epithelial cells. J Immunol 2008; 180: 5689-5698
• 30 Pauley KM, Cha S, Chan EKL. MicroRNA in autoimmunity and autoimmune diseases. J Autoimmun 2009; 32: 189-194
• 31 Nakasa T, Miyaki S, Okubo A et al. Expression of microRNA-146 in rheumatoid arthritis synovial tissue. Arthritis Rheum 2008; 58: 1284-1292
• 32 Chen HF, Hu TT, Zheng XY et al. Association between miR-146a rs2910164 polymorphism and autoimmune diseases susceptibility: a meta-analysis. Gene 2013; 521: 259-264
• 33 Thai T-H, Calado DP, Casola S et al. Regulation of the germinal center response by microRNA-155. Science 2007; 316: 604-608
• 34 Glinsky GV. Phenotype-defining functions of multiple non-coding RNA pathways. Cell Cycle 2008; 7: 1630-1639
• 35 Genome-wide association study of 14 000 cases of seven common diseases and 3 000 shared controls. Nature 2007; 447: 661-678
• 36 Hung C-S, Liu H-H, Liu J-J et al. MicroRNA-200a and -200b Mediated Hepatocellular Carcinoma Cell Migration Through the Epithelial to Mesenchymal Transition Markers. Ann Surg Oncol 2012;
• 37 Soubani O, Ali AS, Logna F et al. Re-expression of miR-200 by novel approaches regulates the expression of PTEN and MT1-MMP in pancreatic cancer. Carcinogenesis 2012;
• 38 Feldkamp J. Autoimmune thyroiditis: diagnosis and treatment. Dtsch Med Wochenschr 2009; 134: 2504-2509