Up-Regulation of Tmevpg1 and Rmrp LncRNA Levels in Splenocytes and Brain of Mouse with Experimental Autoimmune Encephalomyelitis
Background: Two long noncoding (lnc) RNAs, which have been recognized as Tmevpg1/Ifng-AS1/NeST and Rmrp play indispensable roles in the differentiation of TH1 and TH17, respectively. The aim of the present scientific study was to analyze the expression levels of the aforementioned lncRNAs in experimental autoimmune encephalomyelitis (EAE) as an animal model for multiple sclerosis (MS).
Materials and Methods: Initially, EAE was induced in C57BL/6 mice via immunization by using MOG peptide. The leukocyte infiltration rate and demyelination of neuronal axons were determined. Secondly, the expression levels of Tmevpg1, Rmrp, Tbx21, and Rorc were analyzed in the cultured splenocytes and brain lysates, by using Real-Time PCR assay; eventually, the levels of interferon-gamma and interleukin-17 evaluated by ELISA.
Results: Gene expression analysis revealed that Rorc expression in the splenocytes of EAE mice in comparison to the controls was elevated; however, Tbx21 expression did not show any significant difference. Tmevpg1 and Rmrp levels increased in the splenocytes of EAE mice (4.48 times and 39.70 times, respectively, p = 0.0001). Besides, in the brain lysate, the entire genes that have been mentioned were higher than the controls (Tmevpg1: 3.35 times p = 0.02 and Rmrp 11.21 times, p = 0.0001).
Conclusion: The marked up-regulation in Tmevpg1 and Rmrp transcripts suggested the essential roles of lncRNAs in the pathogenesis of EAE and multiple sclerosis indeed. Further investigations are necessary to evaluate the values of these lncRNAs as the target for the therapy or molecular marker for disease monitoring.
Compston A, Coles A. Multiple sclerosis. Lancet (London, England). 2002; 359(9313):1221-31.
Fletcher JM, Lalor SJ, Sweeney CM, Tubridy N, Mills KH. T cells in multiple sclerosis and experimental autoimmune encephalomyelitis. Clinical and experimental immunology. 2010;162(1):1-11.
Langrish CL, Chen Y, Blumenschein WM, Mattson J, Basham B, Sedgwick JD, et al. IL-23 drives a pathogenic T cell population that induces autoimmune inflammation. The Journal of experimental medicine. 2005;201(2):233-40.
Szabo SJ, Kim ST, Costa GL, Zhang X, Fathman CG, Glimcher LH. A novel transcription factor, T-bet, directs Th1 lineage commitment. Cell. 2000;100(6):655-69.
Yang XO, Pappu BP, Nurieva R, Akimzhanov A, Kang HS, Chung Y, et al. T helper 17 lineage differentiation is programmed by orphan nuclear receptors ROR alpha and ROR gamma. Immunity. 2008;28(1):29-39.
Gomez JA, Wapinski OL, Yang YW, Bureau JF, Gopinath S, Monack DM, et al. The NeST long ncRNA controls microbial susceptibility and epigenetic activation of the interferon-gamma locus. Cell. 2013;152(4):743-54.
Huang W, Thomas B, Flynn RA, Gavzy SJ, Wu L, Kim SV, et al. DDX5 and its associated lncRNA Rmrp modulate TH17 cell effector functions. Nature. 2015;528(7583):517-22.
Guttman M, Amit I, Garber M, French C, Lin MF, Feldser D, et al. Chromatin signature reveals over a thousand highly conserved large non-coding RNAs in mammals. Nature. 2009;458(7235):223-7.
Kornienko AE, Guenzl PM, Barlow DP, Pauler FM. Gene regulation by the act of long non-coding RNA transcription. BMC biology. 2013;11:59.
Collier SP, Collins PL, Williams CL, Boothby MR, Aune TM. Cutting edge: influence of Tmevpg1, a long intergenic noncoding RNA, on the expression of Ifng by Th1 cells. Journal of immunology. 2012;189(5):2084-8.
Linder P, Jankowsky E. From unwinding to clamping - the DEAD box RNA helicase family. Nature reviews Molecular cell biology. 2011;12(8):505-16.
Atianand MK, Caffrey DR, Fitzgerald KA. Immunobiology of Long Noncoding RNAs. Annu Rev Immunol. 2017;35:177-98.
Peng H, Liu Y, Tian J, Ma J, Tang X, Rui K, et al. The Long Noncoding RNA IFNG-AS1 Promotes T Helper Type 1 Cells Response in Patients with Hashimoto's Thyroiditis. Scientific reports. 2015;5:17702.
Wang J, Peng H, Tian J, Ma J, Tang X, Rui K, et al. Upregulation of long noncoding RNA TMEVPG1 enhances T helper type 1 cell response in patients with Sjogren syndrome. Immunologic research. 2016;64(2):489-96.
Li H, Hao Y, Zhang D, Fu R, Liu W, Zhang X, et al. Aberrant expression of long noncoding RNA TMEVPG1 in patients with primary immune thrombocytopenia. Autoimmunity. 2016; 49(7):496-502.
Ridanpaa M, van Eenennaam H, Pelin K, Chadwick R, Johnson C, Yuan B, et al. Mutations in the RNA component of RNase MRP cause a pleiotropic human disease, cartilage-hair hypoplasia. Cell. 2001;104(2):195-203.
Feng W, Li L, Xu X, Jiao Y, Du W. Up-regulation of the long non-coding RNA RMRP contributes to glioma progression and promotes glioma cell proliferation and invasion. Archives of medical science: AMS. 2017;13(6):1315-21.
Greco S, Zaccagnini G, Perfetti A, Fuschi P, Valaperta R, Voellenkle C, et al. Long noncoding RNA dysregulation in ischemic heart failure. Journal of translational medicine. 2016;14(1):183.
Zhang F, Gao C, Ma XF, Peng XL, Zhang RX, Kong DX, et al. Expression Profile of Long Noncoding RNAs in Peripheral Blood Mononuclear Cells from Multiple Sclerosis Patients. CNS neuroscience & therapeutics. 2016; 22(4):298-305.
Stromnes IM, Goverman JM. Active induction of experimental allergic encephalomyelitis. Nature protocols. 2006;1(4):1810-9.
Majidi-Zolbanin J, Doosti MH, Kosari-Nasab M, Salari AA. Prenatal maternal immune activation increases anxiety- and depressive-like behaviors in offspring with experimental autoimmune encephalomyelitis. Neuroscience. 2015;294:69-81.
Cai Y, Shen H, Qin C, Zhou J, Lai W, Pan J, et al. The Spatio-Temporal Expression Profiles of CD4 + T Cell Differentiation and Function-Related Genes During EAE Pathogenesis. Inflammation. 2017;40(1):195-204.
Lee J, Choi J, Lee W, Ko K, Kim S. Dehydrodiconiferyl alcohol (DHCA) modulates the differentiation of Th17 and Th1 cells and suppresses experimental autoimmune encephalomyelitis. Molecular immunology. 2015;68(2 Pt B):434-44.
Mandolesi G, Musella A, Gentile A, Grasselli G, Haji N, Sepman H, et al. Interleukin-1beta alters glutamate transmission at purkinje cell synapses in a mouse model of multiple sclerosis. J Neurosci. 2013;33(29):12105-21.
Pfaffl MW, Horgan GW, Dempfle L. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic acids research. 2002;30(9):e36.
Huang W, Littman DR. Regulation of RORgammat in Inflammatory Lymphoid Cell Differentiation. Cold Spring Harbor symposia on quantitative biology. 2015;80:257-63.
Padua D, Mahurkar-Joshi S, Law IK, Polytarchou C, Vu JP, Pisegna JR, et al. A long noncoding RNA signature for ulcerative colitis identifies IFNG-AS1 as an enhancer of inflammation. American journal of physiology Gastrointestinal and liver physiology. 2016;311(3):446-57.
Ivanov, II, McKenzie BS, Zhou L, Tadokoro CE, Lepelley A, Lafaille JJ, et al. The orphan nuclear receptor RORgammat directs the differentiation program of proinflammatory IL-17+ T helper cells. Cell. 2006;126(6):1121-33.
Huh JR, Littman DR. Small molecule inhibitors of RORgammat: targeting Th17 cells and other applications. Eur J Immunol. 2012;42(9):2232-7.
Robinette ML, Fuchs A, Cortez VS, Lee JS, Wang Y, Durum SK, et al. Transcriptional programs define molecular characteristics of innate lymphoid cell classes and subsets. Nature immunology. 2015;16(3):306-17.
He Z, Wang F, Zhang J, Sen S, Pang Q, Luo S, et al. Regulation of Th17 Differentiation by IKKalpha-Dependent and -Independent Phosphorylation of RORgammat. Journal of immunology. 2017;199(3):955-64.
Reboldi A, Coisne C, Baumjohann D, Benvenuto F, Bottinelli D, Lira S, et al. C-C chemokine receptor 6-regulated entry of TH-17 cells into the CNS through the choroid plexus is required for the initiation of EAE. Nature immunology. 2009;10(5):514-23.
Vigneau S, Rohrlich PS, Brahic M, Bureau JF. Tmevpg1, a candidate gene for the control of Theiler's virus persistence, could be implicated in the regulation of gamma interferon. Journal of virology. 2003; 77(10):5632-5638.
Anderson DM, Anderson KM, Chang CL, Makarewich CA, Nelson BR, McAnally JR, et al. A micropeptide encoded by a putative long noncoding RNA regulates muscle performance. Cell. 2015;160(4):595-606.