Integrated Analysis of Differentially Expressed miRNAs and mRNAs in Goat Skin Fibroblast Cells in Response to Orf Virus Infection Reveals That cfa-let-7a Regulates Thrombospondin 1 Expression.

Feng Pang,Xinying Wang,Zhenxing Zhang,Li Du,Zhen Chen,Fengyang Wang,Xiaohong Yang,Mengmeng Zhang,Qi An,Chengqiang Wang

doi:10.3390/v12010118

Abstract

Orf is a zoonotic disease that has caused huge economic losses globally. Systematical analysis of dysregulated cellular micro RNAs (miRNAs) in response to Orf virus (ORFV) infection has not been reported. In the current study, miRNA sequencing and RNA sequencing (RNA-seq) were performed in goat skin fibroblast (GSF) cells at 0, 18, and 30 h post infection (h.p.i). We identified 140 and 221 differentially expressed (DE) miRNAs at 18 and 30 h.p.i, respectively. We also identified 729 and 3961 DE genes (DEGs) at 18 and 30 h.p.i, respectively. GO enrichment analysis indicates enrichment of apoptotic regulation, defense response to virus, immune response, and inflammatory response at both time points. DE miRNAs and DEGs with reverse expression were used to construct miRNA-gene networks. Seven DE miRNAs and seven DEGs related to “negative regulation of viral genome replication” were identified. These were validated by RT-qPCR. Cfa-let-7a, a significantly upregulated miRNA, was found to repress Thrombospondin 1 (THBS1) mRNA and protein expression by directly targeting the THBS1 3′ untranslated region. THBS1 has been reported to induce apoptosis; therefore, the cfa-let-7a-THBS1 axis may play an important role in cellular apoptosis during ORFV infection. This study provides new insights into ORFV and host cell interaction mechanisms.

Highlights

Orf, known as contagious ecthyma, is a zoonotic disease that has led to great economic losses in livestock production globally [1]
We focused on differentially expressed (DE) micro RNAs (miRNAs) and DE genes (DEGs) during the late stage of Orf virus (ORFV) infection—18 and 30 h.p.i
1151 miRNAs were common in all three groups

Summary

Introduction

Known as contagious ecthyma, is a zoonotic disease that has led to great economic losses in livestock production globally [1]. The disease mainly affects goats and sheep, and affects other ruminants and mammals such as musk ox, steenbok, reindeer, dog, and cat [1,2]. People can become infected following contact with infected animals [3]. ORFV, a member of the genus Parapoxvirus, is the causative agent of Orf. ORFV has a double-stranded DNA genome of approximately 130–140 kb, encoding 132 genes [4]. The relatively conserved central regions of the ORFV genome are responsible for morphogenesis and viral replication while the highly variable terminal regions are responsible for virus virulence [5]

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Results

Conclusion