Table_3_Characterization of microRNA Profiles in Pasteurella multocida-Infected Rabbits and Identification of miR-29-5p as a Regulator of Antibacterial Immune Response.XLS

Pasteurella multocida is the pathogenic agent for a variety of severe diseases in livestock, including rabbits. MicroRNAs (miRNAs) participate in the immune response to the pathogen. Distinct miRNA expression patterns were explored in rabbit lung by small-RNA deep sequencing to assess dysregulated miRNAs during P. multocida infection. Totally, 571 miRNAs were screened, of which, 62 were novel, and 32 exhibited differential expression (DE). Of the 32 known DE-miRNAs, 13 and 15 occurred at 1 day and 3 days post-infection (dpi); and ocu-miR-107-3p and ocu-miR-29b-5p were shared between the two time points. Moreover, 7,345 non-redundant target genes were predicted for the 32 DE-miRNAs. Putative target genes were enriched in diverse GO and KEGG pathways and might be crucial for disease resistance. Interestingly, upregulation of ocu-miR-29-5p suppresses P. multocida propagation and downregulates expression of epithelial membrane protein-2 (EMP2) and T-box 4 (TBX4) genes by binding to their 3′ untranslated region in RK13 cells. Thus, ocu-miR-29-5p may indirectly inhibit P. multocida invasion by modulating genes related to the host immune response, such as EMP2 and TBX4.

多杀性巴氏杆菌（Pasteurella multocida）是引发多种家畜（包括家兔）严重疾病的病原菌。微小RNA（miRNAs）参与宿主针对该病原菌的免疫应答反应。为评估多杀性巴氏杆菌感染过程中差异表达的miRNAs，本研究通过小RNA深度测序对家兔肺组织的miRNA表达谱进行了分析。本次研究共筛选得到571个miRNAs，其中62个为新发现的miRNA，另有32个呈现差异表达（DE）。在这32个已知的差异表达miRNAs中，13个与15个分别在感染后1天（dpi）与感染后3天（dpi）被检测到；且ocu-miR-107-3p与ocu-miR-29b-5p在两个时间点均存在差异表达。此外，针对这32个差异表达miRNAs，共预测得到7345个非冗余靶基因。预测得到的靶基因富集于多种基因本体（Gene Ontology，GO）与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）通路，可能在家兔抗病过程中发挥关键作用。值得注意的是，在RK13细胞中，ocu-miR-29-5p的过表达可通过结合上皮膜蛋白2（EMP2）与T-box转录因子4（TBX4）基因的3'非翻译区（3′ untranslated region），抑制多杀性巴氏杆菌的增殖并下调这两个基因的表达。综上，ocu-miR-29-5p可通过调控宿主免疫相关基因（如EMP2与TBX4）的表达，间接抑制多杀性巴氏杆菌的入侵。