Genome-Wide Differential Expression Profiling of Pulmonary circRNAs Associated With Immune Reaction to Pasteurella multocida in Goats

Abstract

Pasteurella multocida is a highly versatile pathogen that infects a wide range of animals, including goats, causing pneumonia and hemorrhagic septicemia. Circular RNA (circRNA) is a type of non-coding RNA that plays an important role in regulating cellular metabolism. However, whether and how circRNA is involved in regulating immune responses in the goat lung has not been reported. Thus, this study was designed to examine the function of circRNA in goats infected with Pasteurella multocida. Goats were assigned into one of two groups: an uninfected control group (CK) and an infected group challenged with P. multocida. Compared with the CK group, which remained healthy, the infected goats showed clinical signs of infection, including depression, cough, nasal discharge, and dyspnea, along with elevated body temperature and lesions in the lung. Whole-transcriptome sequencing and small RNA sequencing were then performed using lung samples from goats from each group. A total of 138 circRNA, 56 microRNAs (miRNA), and 2,673 messenger RNA (mRNA) molecules were significantly differentially expressed in the P. multocida-infected group compared with the CK group. Randomly selected differentially expressed circRNA, miRNA, and mRNA molecules (n = 5 per group) were then validated by quantitative reverse-transcriptase polymerase chain reaction analysis. Gene ontology (GO) analysis of the source genes indicated that six immune-related terms were enriched among the differentially expressed cirRNA molecules, including inflammatory response, immune effector process, cell activation involved in immune response, cytokine-mediated signaling pathway, response to endogenous stimulus, and immune response. The corresponding circRNA molecules were then selected for construction of a competitive endogenous RNA network to identify networks that may be involved in the immune response to P. multocida infection. The results indicated that P. multocida HN01 may cause pneumonia and stimulate an immune response in goats via regulation of circRNA expression. This study presents the first comprehensive circRNA profile in response to P. multocida infection in goats, thus, providing a basis for understanding the function of circRNA in the host immune response to P. multocida infection.

The mRNA and miRNA profiles of goat bronchial epithelial cells stimulated by Pasteurella multocida strains of serotype A and D

“…Wu and coworkers established a P. multocida -induced pneumonia mouse model and transcriptome sequencing showed that the expression of the IL-17 gene was significantly upregulated in the lungs of P. multocida -infected mice ( Wu et al, 2017 ). In our previous study, transcriptome sequencing and small RNA sequencing on goats challenged with P. multocida identified 2,673 significantly differentially expressed mRNAs and 56 miRNAs, some of them are involved in inflammatory response, immune effector process, and cell activation in response to endogenous stimuli ( Chen et al, 2021 ).…”

Effects of Pasteurella multocida on Histopathology, miRNA and mRNA Expression Dynamics in Lung of Goats

“…This study aimed to explore the pathogenic mechanisms of serotype D Pm–infected Hainan black goats. A previous study reported that Pm–challenged goats showed changes at the transcriptome level in the goat lung [ 10 ]. Therefore, this study explored the effects of different infection periods on the miRNA and mRNA levels of goats.…”

“…cp037861). Pm HN01 isolates were revived for the current study as described previously [ 10 ]. Briefly, the bacteria were confirmed by a 16s rRNA polymerase chain reaction (PCR).…”

“…Sequence about the primers and probes is provided in Table A1 . The reaction mixtures used for the qPCR assays were the same as those described by Chen et al [ 10 ]. All reactions were carried out at 95 °C for 5 min, followed by 40 cycles of 95 °C for 15 s and 60 °C for 30 s. The relative expression of the target genes and miRNAs was calculated using the 2 − △△Ct method and normalized to the expression of the house-keeping genes (GAPDH and 5S rRNA).…”

Up-Regulation of Interleukin-10 in Splenic Immune Response Induced by Serotype A Pasteurellamultocida

“…These virulence factors participate in the processes of bacterial growth, colonization, host infection, and cell damage. As the main components of the bacterium, the capsule and LPS play important roles in the pathogenesis of P. multocida [ 5 ]. For example, the virulence of P. multocida capsule mutants in mice was weak [ 5 ]; in the UDP-galactose (galE) mutant of P. multocida , LPS biosynthesis was thoroughly weakened in mice [ 6 ]; the virulence of different serotypes of P. multocida capsule mutants substantially decreased in mice [ 6 ].…”