Background/Purpose:

Primary Sjögren’s syndrome (pSS) is a complex autoimmune disease characterized by a progressive hypofunction of the salivary and lachrymal glands. In order to better understand the pathways potentially involved in the outcome and progression of the disease, we used the latest RNA-Seq technique to characterize the transcriptome of the human salivary gland. Sjögren’s syndrome patients’ salivary gland and compare it to healthy controls.

Methods:

Total RNA was isolated from minor salivary glands stored in RNAlater from healthy volunteers and a primary Sjögren’s syndrome patient. The amount and the quality of the RNA was assessed by different system (Nanodrop, Qubit and Bioanalyzer). 2 mg of total RNA were depleted of the ribosomal RNA, fragmented, retro-transcribed and amplified according to the library construction protocols for the Ion Torrent sequencer of Life Technologies. The library quality and enrichment was assessed by Bioanalyzer and Qubit. The libraries obtained were then loaded in the Ion PI chip and ran on the Ion Torrent semiconductor sequencer, system based on the standard pyrosequencing chemistry. The data generated were then aligned to the human sequence databases (hg19) and analyzed with Partek Suite.

Results:

The average size library was 160 bp, and the final number of aligned sequences to the human genome database was 2.84E+07 (Healthy Volunteers) and 1.36E+07 (pSS patients). These genome-matched reads were then analyzed with Partek Suite and a list of 43.000 mRNAs was screened in order to find expression differences. Preliminary analysis identified several genes differentially expressed in pSS versus controls. Functional analysis identified both well established in autoimmunity and novel pathways that might play a role in Sjogren’s syndrome biogenesis, such as cytokine-cytokine receptor interactions. Analysis of mRNA splicing also identified significant and unexpected differences between pSS and healthy controls.

Conclusion:

This is the first known effort for comparison of the transcriptome of Sjögren’s Syndrome patients’ salivary glands versus healthy controls. Using the latest RNA-Seq technique we were able to identify important pathway and genes differentially expressed that could shed light to the mechanism of salivary gland hypofunction in pSS.

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« Back to 2013 ACR/ARHP Annual Meeting

ACR Meeting Abstracts - https://acrabstracts.org/abstract/comparative-study-of-the-transcriptome-of-minor-salivary-gland-of-sjogrens-syndrome-patients-versus-healthy-controls-based-on-rna-seq/