The Critical Biomarkers Identification of Insulin Signaling Involved in Initiating cAMP Signaling Mediated Salivary Secretion in Sjogren Syndrome: Transcriptome Sequencing in NOD Mice Model

Bo Chen; Jiannan Zhou; Tianjiao Mao; Tingting Cao; Shilin Hu; Wenqi Zhang; Xueyang Li; Xiuni Qin; Xintong Liu; Nobumoto Watanabe; Jiang Li

doi:10.1186/s12575-022-00189-5

The Critical Biomarkers Identification of Insulin Signaling Involved in Initiating cAMP Signaling Mediated Salivary Secretion in Sjogren Syndrome: Transcriptome Sequencing in NOD Mice Model

Biol Proced Online. 2022 Dec 27;24(1):26. doi: 10.1186/s12575-022-00189-5.

Authors

Bo Chen¹, Jiannan Zhou¹, Tianjiao Mao¹, Tingting Cao¹, Shilin Hu¹, Wenqi Zhang², Xueyang Li¹, Xiuni Qin³, Xintong Liu^{4

5}, Nobumoto Watanabe^{4

5}, Jiang Li⁶

Affiliations

¹ Guangdong Engineering Research Center of Oral Restoration and Reconstruction, Affiliated Stomatology Hospital of Guangzhou Medical University, #195 Dongfeng West Road, Guangzhou, 510140, Guangdong, China.
² School of Basic Medicine, Guangzhou Medical University, Guangzhou, Guangdong, China.
³ Guangzhou Concord Cancer Center, Guangzhou, Guangdong, China.
⁴ Chemical Biology Research Group, RIKEN Center for Sustainable Resource Science, Wako, Saitama, 351-0198, Japan.
⁵ Bio-Active Compounds Discovery Unit, RIKEN Center for Sustainable Resource Science, Wako, Saitama, 351-0198, Japan.
⁶ Guangdong Engineering Research Center of Oral Restoration and Reconstruction, Affiliated Stomatology Hospital of Guangzhou Medical University, #195 Dongfeng West Road, Guangzhou, 510140, Guangdong, China. ljiang@gzhmu.edu.cn.

Abstract

Background: Sjogren's syndrome (SS) is an autoimmune disorder characterized by the destruction of exocrine glands, resulting in dry mouth and eyes. Currently, there is no effective treatment for SS, and the mechanisms associated with inadequate salivary secretion are poorly understood.

Methods: In this study, we used NOD mice model to monitor changes in mice's salivary secretion and water consumption. Tissue morphology of the submandibular glands was examined by H&E staining, and Immunohistochemical detected the expression of AQP5 (an essential protein in salivary secretion). Global gene expression profiling was performed on submandibular gland tissue of extracted NOD mice model using RNA-seq. Subsequently, a series of bioinformatics analyses of transcriptome sequencing was performed, including differentially expressed genes (DEGs) identification, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, PPI network construction, hub gene identification, and the validity of diagnostic indicators using the dataset GSE40611. Finally, IFN-γ was used to treat the cells, the submandibular gland tissue of NOD mice model was extracted, and RT-qPCR was applied to verify the expression of hub genes.

Results: We found that NOD mice model had reduced salivary secretion and increased water consumption. H&E staining suggests acinar destruction and basement membrane changes in glandular tissue. Immunohistochemistry detects a decrease in AQP5 immunostaining within acinar. In transcriptome sequencing, 42 overlapping DEGs were identified, and hub genes (REN, A2M, SNCA, KLK3, TTR, and AZGP1) were identified as initiating targets for insulin signaling. In addition, insulin signaling and cAMP signaling are potential pathways for regulating salivary secretion and constructing a regulatory relationship between target-cAMP signaling-salivary secretion.

Conclusion: The new potential targets and signal axes for regulating salivary secretion provide a strategy for SS therapy in a clinical setting.

Keywords: Biomarkers; Insulin signaling; NOD mice model; Salivary secretion; Sjogren syndrome (SS); cAMP signaling.

Abstract

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