Mohammad Amin Honardoost1, Abbas Kiani-Esfahani2, Kamran Ghaedi3, Masoud Etemadifar4, Mansoor Salehi5. 1. Division of Cellular and Molecular Biology, Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran. 2. Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran. 3. Division of Cellular and Molecular Biology, Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran; Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran; Cellular & Molecular Immunology Research Center, Isfahan University of Medical Sciences, Isfahan, Iran. Electronic address: kamranghaedi@yahoo.com. 4. Department of Neurology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran; Multiple Sclerosis and Neuroimmunology Research Center, Isfahan, Iran. 5. Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran. Electronic address: m_salehi@med.mui.ac.ir.
Abstract
BACKGROUND: Multiple sclerosis is an inflammatory autoimmune disease widely characterized by myelin destruction of CNS. Th-17 cells, have been demonstrated to play a crucial role in pathogenesis of MS. MicroRNAs are a new class of non-coding RNAs that participate in post-transcriptional regulation of gene expression. Previous studies have reported a potential role of various miRNAs in induction of Th-17 differentiation and progress of autoimmune diseases. In recent years, it has been shown that miR-326 and miR-26a involved in progress of Th-17 and MS disease. OBJECTIVE: To evaluate expression pattern of miR-326 and miR-26a in peripheral blood lymphocytes of relapsing-remitting MS patients during relapsing and remitting phases compared to healthy control subjects. MATERIALS AND METHODS: Forty RR-MS patients of Isfahan population were diagnosed as relapsing (n=20) or remitting phase (n=20) patients according to clinical manifests and expression level of miR-26a and miR-326 was measured in these groups by quantitative real time PCR method compared to 20 healthy controls. In-silico molecular signaling pathway enrichment analysis was also performed on validated and predicted targets (targetome) of miR-26a by DAVID database to explore possible role of miR-26a in Th17 differentiation. RESULTS: We observed up-regulation of both miR-326 and miR-26a in relapsing phase of multiple sclerosis patients compared with remitting phase (p value=0.0001) and healthy controls (p value=0.0091). ROC curve analysis confirmed valuable and precise potential of miR-326 to discriminate between relapsing and remitting phases of multiple sclerosis with specificity and sensitivity of 100% at a proposed optimum cutoff point. Furthermore, in-silico molecular signaling pathway enrichment analysis detected TGF-β signaling pathway as one of the most statistically relevant pathway with miR-26a targetome. CONCLUSION: Our results confirmed potential of miR-326 as a diagnostic biomarker to discriminate between relapsing and remitting phases of multiple sclerosis disease. Similar expression pattern to miR-326 and in-silico molecular enrichment analysis altogether suggest an inducing role of miR-26a in differentiation of pathogenic Th17 cells during pathogenesis of multiple sclerosis by targeting major components of the TGF-β signaling pathway (i.e. SMAD4 and SMAD1) and disarrangement of this signaling pathway.
BACKGROUND:Multiple sclerosis is an inflammatory autoimmune disease widely characterized by myelin destruction of CNS. Th-17 cells, have been demonstrated to play a crucial role in pathogenesis of MS. MicroRNAs are a new class of non-coding RNAs that participate in post-transcriptional regulation of gene expression. Previous studies have reported a potential role of various miRNAs in induction of Th-17 differentiation and progress of autoimmune diseases. In recent years, it has been shown that miR-326 and miR-26a involved in progress of Th-17 and MS disease. OBJECTIVE: To evaluate expression pattern of miR-326 and miR-26a in peripheral blood lymphocytes of relapsing-remitting MS patients during relapsing and remitting phases compared to healthy control subjects. MATERIALS AND METHODS: Forty RR-MS patients of Isfahan population were diagnosed as relapsing (n=20) or remitting phase (n=20) patients according to clinical manifests and expression level of miR-26a and miR-326 was measured in these groups by quantitative real time PCR method compared to 20 healthy controls. In-silico molecular signaling pathway enrichment analysis was also performed on validated and predicted targets (targetome) of miR-26a by DAVID database to explore possible role of miR-26a in Th17 differentiation. RESULTS: We observed up-regulation of both miR-326 and miR-26a in relapsing phase of multiple sclerosispatients compared with remitting phase (p value=0.0001) and healthy controls (p value=0.0091). ROC curve analysis confirmed valuable and precise potential of miR-326 to discriminate between relapsing and remitting phases of multiple sclerosis with specificity and sensitivity of 100% at a proposed optimum cutoff point. Furthermore, in-silico molecular signaling pathway enrichment analysis detected TGF-β signaling pathway as one of the most statistically relevant pathway with miR-26a targetome. CONCLUSION: Our results confirmed potential of miR-326 as a diagnostic biomarker to discriminate between relapsing and remitting phases of multiple sclerosis disease. Similar expression pattern to miR-326 and in-silico molecular enrichment analysis altogether suggest an inducing role of miR-26a in differentiation of pathogenic Th17 cells during pathogenesis of multiple sclerosis by targeting major components of the TGF-β signaling pathway (i.e. SMAD4 and SMAD1) and disarrangement of this signaling pathway.
Authors: Pernille M Madsen; Dario Motti; Shaffiat Karmally; David E Szymkowski; Kate Lykke Lambertsen; John R Bethea; Roberta Brambilla Journal: J Neurosci Date: 2016-05-04 Impact factor: 6.167