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

• Published in: PLoS neglected tropical diseases Vol. 16; no. 5; p. e0010379
• Main Authors: Gao, Zheng-Rong, Liu, Qiong, Zhao, Jie, Zhao, Ya-Qiong, Tan, Li, Zhang, Shao-Hui, Zhou, Ying-Hui, Chen, Yun, Guo, Yue, Feng, Yun-Zhi
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 01-05-2022; Public Library of Science (PLoS)
• Subjects: Analysis; Analytical methods; Animals; Bacterial infections; Binding sites; Biology and Life Sciences; Biomedical materials; Bone cells; Bone growth; Bone remodeling; Bone remodelling; Bones; Circular RNA; CLOCK protein; Diagnosis; Differentiation (biology); Encyclopaedias; Encyclopedias; Ethics; Gene expression; Gene Expression Profiling; Gene Regulatory Networks; Genes; Genetic aspects; Genomes; Health aspects; Immunohistochemistry; Infections; Leprosy; Medical research; Medicine and Health Sciences; Mesenchyme; Messenger RNA; Mice; MicroRNA; MicroRNAs; MicroRNAs - genetics; MicroRNAs - metabolism; miRNA; Muramyl dipeptide; Mycobacterium leprae; Mycobacterium leprae - genetics; Mycobacterium leprae - metabolism; Nucleic acids; Nucleotide sequence; Osteocytes; Osteocytes - metabolism; Osteogenesis; Osteogenesis - genetics; PCR; Polymerase chain reaction; Proteins; Research and Analysis Methods; Ribonucleic acid; RNA; RNA, Circular - genetics; RNA, Messenger - genetics; RNA, Messenger - metabolism; Stem cell transplantation; Stem cells; Transcription; Tropical diseases; Western blotting; China; United States > US
• ISSN: 1935-2735; 1935-2727; 1935-2735
• DOI: 10.1371/journal.pntd.0010379

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.
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.
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.