Regulation of the Human IL-10RB Gene Expression by Sp8 and Sp9

Article type: Research Article

Authors: Luo, Shuyue^a | Dong, Xiangjun^a | Guo, Shipeng^a | Wang, Qunxian^a | Dai, Xi^a | Jiang, Yanshuang^a | Zhu, Weiyi^a | Zhou, Weihui^{a; *} | Song, Weihong^{a; b; c; *}

Affiliations: [a] Chongqing Key Laboratory of Translational Medical Research in Cognitive Development and Learning and Memory Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, National Clinical Research Center for Child Health and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Children’s Hospital of Chongqing Medical University, Chongqing, China | [b] Institute of Aging, Key Laboratory of Alzheimer’s Disease of Zhejiang Province, Zhejiang Provincial Clinical Research Center for Mental Disorders, School of Mental Health and Kangning Hospital, The Second Affiliated Hospital and Yuying Children’s Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, China | [c] Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision and Brain Health), Wenzhou, Zhejiang, China

Correspondence: [*] Correspondence to: Dr. Weihong Song, Chongqing Key Laboratory of Translational Medical Research in Cognitive Development and Learning and Memory Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, National Clinical Research Center for Child Health and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Children’s Hospital of Chongqing Medical University, Chongqing, China. Tel.: +86 577 86699670; E-mail: [email protected]; Dr. Weihui Zhou, Tel.: +23 63637854; E-mail: [email protected].

Abstract: Background:Interleukin-10 (IL-10) is a classic anti-inflammatory cytokine that exerts its effects via the receptor complexes IL-10RA and IL-10RB. Loss of IL-10RB results in many diseases. Moreover, IL-10RB is closely associated with neuronal survival and synaptic formation. However, the regulation of IL-10RB gene expression remains elusive. Objective:To investigate whether the expression of IL-10RB gene is increased in brain of Alzheimer’s disease (AD) and its transcriptional regulation. Methods:We examined the gene expression of AD patient brain from public database and detected the protein expression of AD model mouse brain by western blot. We constructed a variety of reporter gene plasmids with different lengths or mutation sites, tested the promoter activity and defined the functional region of the promoter with the luciferase reporter assay. The protein-DNA binding between transcription factors and the promoter was analyzed using chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assay (EMSA). Results:We found that the IL-10RB is elevated in the brain of AD patient and AD model mice. The minimal promoter of the IL-10RB gene is located in the –90 to +51 bp region (relative to the transcriptional start site) and is sufficient for high-level expression of the IL-10RB gene. Transcription factors Sp8 and Sp9 bind to the IL-10RB promoter in vitro. The overexpression or knockdown of Sp8 and Sp9 affected the IL-10RB promoter activity and its gene expression. Conclusion:Our study functionally characterized the promoter of the IL-10RB gene and demonstrated that Sp8 and Sp9 regulated its expression.

Keywords: Gene expression, IL-10RB, Sp8, Sp9, transcriptional regulation

DOI: 10.3233/JAD-220321

Journal: Journal of Alzheimer's Disease, vol. 88, no. 4, pp. 1469-1485, 2022