Integrated mRNA-seq and miRNA-seq analysis of goat fibroblasts response to Brucella Melitensis strain M5-90

Baobao Li; Si Chen; Chengqiang Wang; Qiaoling Chen; Churiga Man; Qi An; Zhenxing Zhang; Zhiyong Liu; Li Du; Fengyang Wang

doi:10.7717/peerj.11679

Integrated mRNA-seq and miRNA-seq analysis of goat fibroblasts response to Brucella Melitensis strain M5-90

PeerJ. 2021 Jun 29:9:e11679. doi: 10.7717/peerj.11679. eCollection 2021.

Authors

Baobao Li^#¹, Si Chen^#¹, Chengqiang Wang¹, Qiaoling Chen¹, Churiga Man¹, Qi An¹, Zhenxing Zhang¹, Zhiyong Liu¹, Li Du¹, Fengyang Wang¹

Affiliation

¹ Hainan Key Lab of Tropical Animal Reproduction, Breeding and Epidemic Disease Research, Animal Genetic Engineering Key Lab of Haikou, College of Animal Science and Technology, Hainan University, Hainan Key Lab of Tropical Animal Reproduction, Haikou, Hainan, China.

^# Contributed equally.

Abstract

Brucellosis is a globally zoonotic bacterial disease of humans and various animals including goats, sheep, and cattle. Brucella melitensis M5-90, a live attenuated vaccine strain, has been widely used to prevent brucellosis in goats and sheep. However, the molecular mechanisms governing protective immunity response in non-professional phagocytes infected with B. melitensis M5-90 have not been fully investigated, especially in goats. In our research, goat fibroblasts were used as in vitro models to determine these mechanisms by transcriptome analysis. After incubating with B. melitensis M5-90 3 h, the infected goat fibroblasts were collected at 0 h, 4 h, 24 h, 48 h and 72 h for RNA-seq. The results indicated that there were totally 11,819 differentially expressed genes (DEGs) and 777 differentially expressed (DE) miRNAs found in experiment groups compared with the control groups (|log2(Foldchange)|≥1, FDR<0.05). GO and KEGG enrichment analyses revealed that down-regulated genes were involved in the riboflavin metabolism and positive regulation of IL-8 secretion pathway. The up-regulated genes were mainly involved in adaptive immunity, including TNF signaling pathway, MAPK signaling pathway and JAK/STAT pathway. Additionally, cytokine-cytokine receptor interaction, natural killer cell mediated cytotoxicity and toll-like receptor signaling pathway, which associated with innate immunity pathways, were also induced. Based on the Pearson correlation coefficients and prediction results of TargetScan and miRanda, the miRNA-mRNA networks of NFKB1, IFNAR2 and IL10RB were constructed and verified in goat fibroblasts by qPCR, which demonstrated that goat fibroblasts displayed immunomodulatory properties. Our findings provide a deeper insight into the host miRNA-driven B. melitensis defense mechanism and reveal the transcriptome changes involved in the innate and adaptive immune response of the goats to B. melitensis infection.

Keywords: Adaptive immunity; Brucella melitensis strain M5-90; Goat fibroblasts; Innate immunity; Transcriptomics.

Grants and funding

This work was financially supported by Natural Science Foundation of China (No. U1803236), Hainan Provincial Natural Science Foundation of China (No. 2019RC123) and Scientific Research Foundation of Hainan University (No. KYQD (ZR) 1946). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.