Transcriptome analysis reveals the mechanism of stromal cell-derived factor-1 and exendin-4 synergistically promoted periodontal ligament stem cells osteogenic differentiation

Wenyan Kang; Lingqian Du; Qianyu Liang; Rui Zhang; Chunxu Lv; Shaohua Ge

doi:10.7717/peerj.12091

Transcriptome analysis reveals the mechanism of stromal cell-derived factor-1 and exendin-4 synergistically promoted periodontal ligament stem cells osteogenic differentiation

PeerJ. 2021 Aug 27:9:e12091. doi: 10.7717/peerj.12091. eCollection 2021.

Authors

Wenyan Kang¹, Lingqian Du², Qianyu Liang¹, Rui Zhang¹, Chunxu Lv¹, Shaohua Ge¹

Affiliations

¹ Department of Periodontology, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, Shandong, China.
² Department of Stomatology, The Second Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China.

Abstract

Stromal cell-derived factor-1 (SDF-1) and Exendin-4 (EX-4) play beneficial roles in promoting periodontal ligament stem cells (PDLSCs) osteogenic differentiation, while the detailed mechanism has not been clarified. In this study, we aimed to evaluate the biological mechanism of SDF-1 and EX-4 alone or synergistic application in regulating PDLSCs differentiation by RNA-sequencing (RNA-seq). A total of 110, 116 and 109 differentially expressed genes (DEGs) were generated in osteogenic medium induced PDLSCs treated by SDF-1, EX-4, and SDF-1+EX-4, respectively. The DEGs in SDF-1 group were enriched in signal transduction related signaling pathways; the DEGs in EX-4 group were enriched in metabolism and biosynthesis-related pathways; and the DEGs generated in SDF-1+EX-4 group were mainly enriched in RNA polymerase II transcription, cell differentiation, chromatin organization, protein phosphorylation pathways. Based on Venn analysis, a total of 37 specific DEGs were identified in SDF-1+EX-4 group, which were mainly enriched in negative regulation of autophagy and cellular component disassembly signaling pathways. Short time-series expression miner (STEM) analysis grouped all expressed genes of PDLSCs into 49 clusters according to the dynamic expression patterns and 25 genes, including NRSN2, CHD9, TUBA1A, distributed in 10 gene clusters in SDF-1+EX-4 treated PDLSCs were significantly up-regulated compared with the SDF-1 and EX-4 alone groups. The gene set enrichment analysis indicated that SDF-1 could amplify the role of EX-4 in regulating varied signaling pathways, such as type II diabetes mellitus and insulin signaling pathways; while EX-4 could aggravate the effect of SDF-1 on PDLSCs biological roles via regulating primary immunodeficiency, tight junction signaling pathways. In summary, our study confirmed that SDF-1 and EX-4 combined application could enhance PDLSCs biological activity and promote PDLSCs osteogenic differentiation by regulating the metabolism, biosynthesis and immune-related signaling pathways.

Keywords: Exendin-4; Osteogenic differentiation; Periodontal ligament stem cells; RNA-seq; Stromal cell-derived factor-1.

Grants and funding

This research was supported by the Shandong Provincial Natural Science Foundation (No. ZR2020QH159), the National Natural Science Foundation of China (No. 81670993, 81873716, 82170964 and 8210033710), The Construction Engineering Special Fund of “Taishan Scholars” of Shandong Province (No. ts20190975 and tsqn201909180), Rongxiang Regenerative Medicine Foundation of Shandong University (No. 2019SDRX-14), National Key R&D Program of China (No. 2017YFB0405400), Collaborative Innovation Center of Technology and Equipment for Biological Diagnosis and Therapy in Universities of Shandong, The National Key Research and Development Program of China (No. 2017YFA0104604), Open Foundation of Shandong Provincial Key Laboratory of Oral Tissue Regeneration (No. SDKQ201901, SDKQ201904), and The Youth scientific research funds of school of Stomatology, Shandong university (2020QNJJ01). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.