title: Disulfiram and Diphenhydramine Hydrochloride Upregulate miR-30a to Suppress IL-17-Associated Autoimmune Inflammation.
abstract: T-helper 17 (Th17) cells play an important role in the pathogenesis of multiple sclerosis (MS), an autoimmune demyelinating disease that affects the CNS. In the present study, MicroRNA sequencing (miRNA-seq) was performed in mouse Th0 and Th17 cells to determine the critical miRNAs that are related to Th17 differentiation. We found that miR-30a was significantly downregulated during mouse Th17 differentiation. In addition, the level of miR-30a in CD4(+) T cells from peripheral blood of MS patients and experimental autoimmune encephalomyelitis (EAE) animal models was also decreased and inversely correlated with the expression of interleukin 17a, the canonical cytokine of Th17 cells. Moreover, overexpression of miR-30a inhibited Th17 differentiation and prevented the full development of EAE, whereas interference of miR-30a promoted Th17 differentiation. Mechanism studies showed that miR-30a reduced IRF4 expression by specifically binding with the 3'-untranslated region. Through screening of 640 different Food and Drug Administration (FDA)-approved drugs, we found that disulfiram and diphenhydramine hydrochloride were effective candidates for inhibiting Th17 differentiation and ameliorating EAE development through upregulating miR-30a. To our knowledge, the present work is not only the first miRNA-seq study focusing on Th17 differentiation, but also the first chemical screening for FDA-approved drugs that inhibit Th17 differentiation through regulating miRNA expression.
  
  
  
  The present work is the first miRNA sequencing (miRNA-seq) study focusing on T-helper 17 (Th17) differentiation. By miRNA deep sequencing, we found that miR-30a was downregulated during Th17 differentiation. miR-30a was also decreased in CD4(+) T cells from multiple sclerosis patients and experimental autoimmune encephalomyelitis (EAE) mice. miR-30a reduced IRF4 expression by specific binding with the 3'-untranslated region and thus suppressed Th17 differentiation and prevented the full development of EAE. Interestingly, by performing a chemical screen with Food and Drug Administration-approved small-molecule drugs, we found that disulfiram and diphenhydramine upregulated miR-30a and suppressed Th17-associated autoimmune demyelination.
authors: Institute of Neuroscience, Key Laboratory of Molecular Neurobiology of the Ministry of Education and the Collaborative Innovation Center for Brain Science, Second Military Medical University, Shanghai 200433, China, Department of Neurology, Chinese PLA 254 Hospital, Tianjin, China.
authors: Ming
authors: M
authors: Zhao
authors: Institute of Neuroscience, Key Laboratory of Molecular Neurobiology of the Ministry of Education and the Collaborative Innovation Center for Brain Science, Second Military Medical University, Shanghai 200433, China.
authors: Dingya
authors: D
authors: Sun
authors: Neurology Department, Renji Hospital, Shanghai 200127, China.
authors: Yangtai
authors: Y
authors: Guan
authors: Institute of Neuroscience, Key Laboratory of Molecular Neurobiology of the Ministry of Education and the Collaborative Innovation Center for Brain Science, Second Military Medical University, Shanghai 200433, China.
authors: Zhihong
authors: 0000-0001-7037-8488
authors: Z
authors: Wang
authors: Neurology Department, the First Affiliated Hospital of Bengbu Medical College, Anhui 233004, China.
authors: Daoqian
authors: D
authors: Sang
authors: Institute of Neuroscience, Key Laboratory of Molecular Neurobiology of the Ministry of Education and the Collaborative Innovation Center for Brain Science, Second Military Medical University, Shanghai 200433, China.
authors: Mingdong
authors: 0000-0003-2045-4969
authors: M
authors: Liu
authors: Institute of Neuroscience, Key Laboratory of Molecular Neurobiology of the Ministry of Education and the Collaborative Innovation Center for Brain Science, Second Military Medical University, Shanghai 200433, China.
authors: Yingyan
authors: Y
authors: Pu
authors: Institute of Neuroscience, Key Laboratory of Molecular Neurobiology of the Ministry of Education and the Collaborative Innovation Center for Brain Science, Second Military Medical University, Shanghai 200433, China.
authors: Xue
authors: X
authors: Fang
authors: Institute of Neuroscience, Key Laboratory of Molecular Neurobiology of the Ministry of Education and the Collaborative Innovation Center for Brain Science, Second Military Medical University, Shanghai 200433, China.
authors: Dan
authors: D
authors: Wang
authors: Institute of Neuroscience, Key Laboratory of Molecular Neurobiology of the Ministry of Education and the Collaborative Innovation Center for Brain Science, Second Military Medical University, Shanghai 200433, China.
authors: Aijun
authors: A
authors: Huang
authors: Neurology Department, Changhai Hospital, Shanghai 200433, China, and.
authors: Xiaoying
authors: X
authors: Bi
authors: Institute of Neuroscience, Key Laboratory of Molecular Neurobiology of the Ministry of Education and the Collaborative Innovation Center for Brain Science, Second Military Medical University, Shanghai 200433, China, caoli@smmu.edu.cn chenghe@smmu.edu.cn.
authors: Li
authors: L
authors: Cao
authors: Institute of Neuroscience, Key Laboratory of Molecular Neurobiology of the Ministry of Education and the Collaborative Innovation Center for Brain Science, Second Military Medical University, Shanghai 200433, China, caoli@smmu.edu.cn chenghe@smmu.edu.cn.
authors: Cheng
authors: C
authors: He
mesh_terms: D000818:Animals; D016022:Case-Control Studies; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D004155:Diphenhydramine / Q000494:pharmacology*; D004195:Disease Models, Animal; D004221:Disulfiram / Q000494:pharmacology*; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000378:metabolism* / Q000517:prevention & control; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D050835:Interferon Regulatory Factors / Q000378:metabolism; D020381:Interleukin-17 / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000378:metabolism*; D018991:Myelin Proteolipid Protein / Q000633:toxicity; D063308:Myelin-Oligodendrocyte Glycoprotein / Q000633:toxicity; D010446:Peptide Fragments / Q000633:toxicity; D018709:Statistics, Nonparametric; D014162:Transfection; D015854:Up-Regulation / Q000187:drug effects*; D000099218:Interferon Regulatory Factor-4
chemical_list: D004155:Diphenhydramine; D004221:Disulfiram; D050835:Interferon Regulatory Factors; D020381:Interleukin-17; D035683:MicroRNAs; D018991:Myelin Proteolipid Protein; D063308:Myelin-Oligodendrocyte Glycoprotein; D010446:Peptide Fragments; D000099218:Interferon Regulatory Factor-4; C508717:IL17A protein, human; C534715:Mirn30d microRNA, mouse; C445081:myelin oligodendrocyte glycoprotein (35-55); C435023:myelin proteolipid protein (139-151)
keywords: EAE; FDA-approved drugs; Th17 cells; miRNAs; multiple sclerosis