AB0180 Identification of lncrna lncmkln1 contributed to abnormal activation of type i interferon pathway in systemic lupus erythematosus. (12th June 2018)

• Record Type: Journal Article
• Authors: Ye, Z.; Tang, Y.; Yin, Z.; Chen, X.; Chen, Y.
• Is Part Of: Annals of the rheumatic diseases. Volume 77(2018)Supplement 2
• Page Start: 1277
• Page End: 1278
• Publication Date: 2018-06-12
• Subjects: Rheumatism -- Periodicals; 616.723005
• DOI: 10.1136/annrheumdis-2018-eular.6029
• Languages: English
• ISSNs: 0003-4967

Abstract

Abstract : Background: Dysregulation or dysfunction of some key moleculars in signalling pathway is involved in disease pathogenesis. Type I interferon pathway abnormal activation has been identified as major molecular phenotype of lupus patients. Long non-coding RNA (lncRNA), as a regulator of gene expression, plays great role in signalling pathway. Objectives: We hypothesised that dysregulation of lncRNA may involve in key signalling pathway of SLE. Methods: Deep sequencing of human renal samples to screen differential expression of lncRNAs between LN patients and healthy donors. RNA-FISH was used to identify subcellular location of lncRNA. Stimulation in human renal mesangial cells (HRMC) by all kinds of TLR ligands, IFNs, and TNFα, and transfection in HRMC cells by antisense oligonucleotides (ASOs), and quantitative real-time polymerase chain reaction (RT-qPCR), enzyme linked immunosorbent assay (ELISA) were used to analyse the relative genes expression. LncMKLN1 transcription was activated or inhibited through CRISPR-dCas9 system in Hela cell line. RNA-seq was executed to examine the gene expression profile after changing lncMKLN1 expression, and western blot was applied to determine the key signalling molecules of IFN pathway. Results: LncMKLN1, dominantly located in nucleus, was up-regulated in lupus patients compared to healthy donors, and could be induced by IFNα and TLR ligands in HRMC. Silencing lncMKLN1 significantly reduced the expression of a group of interferon-inducible genes, including IFIT3, OAS1, CXCL10, etc. We used lose-of-function and gain-of-function strategy through CRSIPR system to confirm that lncMKLN1 positively regulated type I interferon pathway. Furthermore, it was identified the involvement of lncMKLN1 in interferon signalling pathway was through regulating the expression of STAT1, IRF9 and phosphorylation of IRF9 and STAT1 although its mechanism is also needed to investigate. Conclusions: Upregulated lncMKLN1 expression contributed to abnormal activation of interferon pathway of SLE. Disclosure of Interest: None declared