Chuammitri Phongsakorn, Vannamahaxay Soulasack, Sornpet Benjaporn, Pringproa Kidsadagon, Patchanee Prapas
Department of Veterinary Biosciences and Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand.
Center of Excellence in Veterinary Biosciences (CEVB), Chiang Mai University, Chiang Mai, Thailand.
PeerJ. 2020 Feb 12;8:e8522. doi: 10.7717/peerj.8522. eCollection 2020.
MicroRNAs (miRNAs) play an essential role in gene regulators in many biological and molecular phenomena. Unraveling the involvement of miRNA as a key cellular factor during in vitro canine parvovirus (CPV) infection may facilitate the discovery of potential intervention candidates. However, the examination of miRNA expression profiles in CPV in tissue culture systems has not been fully elucidated.
In the present study, we utilized high-throughput small RNA-seq (sRNA-seq) technology to investigate the altered miRNA profiling in miRNA libraries from uninfected (Control) and CPV-2c infected Crandell Reese Feline Kidney cells.
We identified five of known miRNAs (miR-222-5p, miR-365-2-5p, miR-1247-3p, miR-322-5p and miR-361-3p) and three novel miRNAs (Novel 137, Novel 141 and Novel 102) by sRNA-seq with differentially expressed genes in the miRNA repertoire of CPV-infected cells over control. We further predicted the potential target genes of the aforementioned miRNAs using sequence homology algorithms. Notably, the targets of miR-1247-3p exhibited a potential function associated with cellular defense and humoral response to CPV. To extend the probing scheme for gene targets of miR-1247-3p, we explored and performed Gene Ontology (GO) enrichment analysis of its target genes. We discovered 229 putative targets from a total of 38 enriched GO terms. The top over-represented GO enrichment in biological process were lymphocyte activation and differentiation, marginal zone B cell differentiation, negative regulation of cytokine production, negative regulation of programed cell death, and negative regulation of signaling. We next constructed a GO biological process network composed of 28 target genes of miR-1247-3p, of which, some genes, namely , , , , , and were among the genes with obviously intersected in multiple GO terms.
The miRNA-1247-3p and its cognate target genes suggested their great potential as novel therapeutic targets or diagnostic biomarkers of CPV or other related viruses.
微小RNA（miRNA）在许多生物学和分子现象的基因调控中发挥着重要作用。阐明miRNA作为关键细胞因子在犬细小病毒（CPV）体外感染过程中的作用，可能有助于发现潜在的干预靶点。然而，在组织培养系统中对CPV感染时miRNA表达谱的研究尚未完全阐明。
在本研究中，我们利用高通量小RNA测序（sRNA-seq）技术，研究未感染（对照）和CPV-2c感染的Crandell Reese猫肾细胞的miRNA文库中miRNA谱的变化。
通过sRNA-seq，我们在CPV感染细胞的miRNA库中鉴定出5个已知miRNA（miR-222-5p、miR-365-2-5p、miR-1247-3p、miR-322-5p和miR-361-3p）和3个新miRNA（新137、新141和新102），其差异表达基因相对于对照有所不同。我们进一步使用序列同源性算法预测了上述miRNA的潜在靶基因。值得注意的是，miR-1247-3p的靶标表现出与细胞对CPV的防御和体液反应相关的潜在功能。为了扩展对miR-1247-3p基因靶标的探究方案，我们对其靶基因进行了基因本体论（GO）富集分析。我们从总共38个富集的GO术语中发现了229个推定靶标。在生物学过程中，最显著的GO富集是淋巴细胞激活和分化、边缘区B细胞分化、细胞因子产生的负调控、程序性细胞死亡的负调控以及信号传导的负调控。接下来，我们构建了一个由miR-1247-3p的28个靶基因组成的GO生物学过程网络，其中，一些基因，即 、 、 、 、 、 和 是在多个GO术语中明显相交的基因。
miRNA-1247-3p及其同源靶基因表明它们作为CPV或其他相关病毒的新型治疗靶点或诊断生物标志物具有巨大潜力。
