A comprehensive analysis of the circRNA-miRNA-mRNA network in osteocyte-like cell associated with Mycobacterium leprae infection

Zheng-Rong Gao; Qiong Liu; Jie Zhao; Ya-Qiong Zhao; Li Tan; Shao-Hui Zhang; Ying-Hui Zhou; Yun Chen; Yue Guo; Yun-Zhi Feng

doi:10.1371/journal.pntd.0010379

A comprehensive analysis of the circRNA-miRNA-mRNA network in osteocyte-like cell associated with Mycobacterium leprae infection

PLoS Negl Trop Dis. 2022 May 2;16(5):e0010379. doi: 10.1371/journal.pntd.0010379. eCollection 2022 May.

Authors

Affiliations

¹ Department of Stomatology, The Second Xiangya Hospital, Central South University, Changsha, Hunan, China.
² National Clinical Research Center for Metabolic Diseases, Hunan Provincial Key Laboratory of Metabolic Bone Diseases, and Department of Metabolism and Endocrinology, The Second Xiangya Hospital of Central South University, Changsha, Hunan, China.

Abstract

Background: Bone formation and loss are the characteristic clinical manifestations of leprosy, but the mechanisms underlying the bone remodeling with Mycobacterium leprae (M. leprae) infection are unclear.

Methodology/principal findings: Osteocytes may have a role through regulating the differentiation of osteogenic lineages. To investigate osteocyte-related mechanisms in leprosy, we treated osteocyte-like cell with N-glycosylated muramyl dipeptide (N.g MDP). RNA-seq analysis showed 724 differentially expressed messenger RNAs (mRNAs) and 724 differentially expressed circular RNA (circRNAs). Of these, we filtered through eight osteogenic-related differentially expressed genes, according to the characteristic of competing endogenous RNA, PubMed databases, and bioinformatic analysis, including TargetScan, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes. Based on these results, we built a circRNA-microRNA (miRNA)-mRNA triple network. Quantitative reverse-transcription polymerase chain reaction and western blots analyses confirmed decreased Clock expression in osteocyte-like cell, while increased in bone mesenchymal stem cells (BMSCs), implicating a crucial factor in osteogenic differentiation. Immunohistochemistry showed obviously increased expression of CLOCK protein in BMSCs and osteoblasts in N.g MDP-treated mice, but decreased expression in osteocytes.

Conclusions/significance: This analytical method provided a basis for the relationship between N.g MDP and remodeling in osteocytes, and the circRNA-miRNA-mRNA triple network may offer a new target for leprosy therapeutics.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Gene Expression Profiling
Gene Regulatory Networks
Leprosy*
Mice
MicroRNAs* / genetics
MicroRNAs* / metabolism
Mycobacterium leprae / genetics
Mycobacterium leprae / metabolism
Osteocytes / metabolism
Osteogenesis / genetics
RNA, Circular / genetics
RNA, Messenger / genetics
RNA, Messenger / metabolism

Substances

MicroRNAs
RNA, Circular
RNA, Messenger

Grants and funding

This study was supported by the National Natural Science Foundation of China (Grants 81773339) to YZF and (Grants 81800788) to YG; the Science and Technology Department of Hunan Province, China (Grants 2017WK2041) to YZF and (Grants 2018SK52511) to YG; the fund for Xiangya Clinical Medicine Database of Central South University (Grant 2014-ZDYZ-1-16) to YZF, and the Open Sharing Fund for the Large-scale Instruments and Equipment of Central South University to YG. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.