Identification of Disease-Susceptible Lncrna Contributed to Abnormal Activation of Type I Interferon Pathway in Systemic Lupus Erythematosus

Nan Shen^1,2, Yuanjia Tang³, Zhixin Xue³ and Chaojie Cui⁴, ¹Shanghai Institute of Rheumatology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China, Shanghai, China, ²The Center for Autoimmune Genomics and Etiology (CAGE), Cincinnati Children’s Hospital Medical Center, Cincinnati, Ohio, United States of America, Cincinnati, OH, ³Shanghai Institute of Rheumatology,Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China, ⁴Shanghai Institute of Rheumatology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

Meeting: 2017 ACR/ARHP Annual Meeting

Date of first publication: September 18, 2017

Keywords: interferons and polymorphism, RNA, SLE

Session Information

Date: Sunday, November 5, 2017

Title: Genetics, Genomics and Proteomics Poster I

Session Type: ACR Poster Session A

Session Time: 9:00AM-11:00AM

Background/Purpose:

Dysregulation or dysfunction of some key moleculars in signaling pathway is involved in disease pathogenesis. Long non-coding RNA (lncRNA), as a regulator of gene expression, plays great role in signaling pathway. The majority of susceptible single nucleotide polymorphisms (SNPs) of systemic lupus erythematosus (SLE) identified in GWAS studies are located in noncoding regions of the human genome. We hypothesized that disease-related functional SNP linked to lncRNA may affect expression or function of lncRNA and involve in key signaling pathway of SLE.

Methods:

Deep sequencing of human renal samples to screen differential expression of lncRNAs between LN patients and healthy donors and mining GWAS data were applied to get candidate lncRNAs. RNA-FISH was used to identify subcellular location of lncRNA. Reporter gene assay was applied to ascertain effect of disease-related SNP on lncMKLN1 transcription. 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) were used to analyze 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 signaling 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. GWAS data showed that there were two lupus-related SNPs located within proximal region of lncMKLN1 promoter. Promoter constructs containing susceptible allele have higher activity of transcription compared to another allele. 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 signaling pathway was through regulating the expression of STAT1, IRF9 and phosphorylation of IRF9 and STAT1 although its mechanism is also needed to investigate.

Conclusion:

Functional susceptible SNP enhances lncMKLN1 expression, resulting in abnormal activation of interferon pathway of SLE.

Disclosure: N. Shen, None; Y. Tang, None; Z. Xue, None; C. Cui, None.

To cite this abstract in AMA style:

Shen N, Tang Y, Xue Z, Cui C. Identification of Disease-Susceptible Lncrna Contributed to Abnormal Activation of Type I Interferon Pathway in Systemic Lupus Erythematosus [abstract]. Arthritis Rheumatol. 2017; 69 (suppl 10). https://acrabstracts.org/abstract/identification-of-disease-susceptible-lncrna-contributed-to-abnormal-activation-of-type-i-interferon-pathway-in-systemic-lupus-erythematosus/. Accessed .

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