miRNA let-7 family regulated by NEAT1 and ARID3A/NF-κB inhibits PRRSV-2 replication in vitro and in vivo

Xiangbin You; Min Liu; Qian Liu; Huijuan Li; Yilin Qu; Xiaoxiao Gao; Chengyu Huang; Gan Luo; Gang Cao; Dequan Xu

doi:10.1371/journal.ppat.1010820

miRNA let-7 family regulated by NEAT1 and ARID3A/NF-κB inhibits PRRSV-2 replication in vitro and in vivo

PLoS Pathog. 2022 Oct 10;18(10):e1010820. doi: 10.1371/journal.ppat.1010820. eCollection 2022 Oct.

Authors

Xiangbin You^{1

2

3

4}, Min Liu^{2

5}, Qian Liu^{2

4

6}, Huijuan Li^{2

3

6}, Yilin Qu^{2

5}, Xiaoxiao Gao^{2

5}, Chengyu Huang^{2

5}, Gan Luo^{2

3

6}, Gang Cao^{1

5}, Dequan Xu^{2

3

6}

Affiliations

¹ State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, China.
² Key Laboratory of Swine Genetics and Breeding of Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan, China.
³ College of Animal Science & Technology, Huazhong Agricultural University, Wuhan, China.
⁴ College of Animal Science & Technology, Henan University of Science and Technology, Luoyang, China.
⁵ College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.
⁶ Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan, China.

Abstract

Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating diseases affecting the swine industry worldwide. To investigate the role of miRNAs in the infection and susceptibility of PRRS virus (PRRSV), twenty-four miRNA libraries were constructed and sequenced from PRRSV-infected and mock-infected Porcine alveolar macrophages (PAMs) of Meishan, Landrace, Pietrain and Qingping pigs at 9 hours post infection (hpi), 36 hpi, and 60 hpi. The let-7 family miRNAs were significantly differentially expressed between PRRSV-infected and mock-infected PAMs from 4 pig breeds. The let-7 family miRNAs could significantly inhibit PRRSV-2 replication by directly targeting the 3'UTR of the PRRSV-2 genome and porcine IL6, which plays an important role in PRRSV replication and lung injury. NEAT1 acts as a competing endogenous lncRNA (ceRNA) to upregulate IL6 by attaching let-7 in PAMs. EMSA and ChIP results confirmed that ARID3A could bind to the promoter region of pri-let-7a/let-7f/let-7d gene cluster and inhibit the expression of the let-7 family. Moreover, the NF-κB signaling pathway inhibits the expression of the let-7 family by affecting the nuclear import of ARID3A. The pEGFP-N1-let-7 significantly reduced viral infections and pathological changes in PRRSV-infected piglets. Taken together, NEAT1/ARID3A/let-7/IL6 play significant roles in PRRSV-2 infection and may be promising therapeutic targets for PRRS.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

3' Untranslated Regions
Animals
DNA-Binding Proteins / genetics
Interleukin-6 / metabolism
Macrophages, Alveolar / metabolism
MicroRNAs* / genetics
MicroRNAs* / metabolism
NF-kappa B / genetics
NF-kappa B / metabolism
Porcine Reproductive and Respiratory Syndrome* / genetics
Porcine Reproductive and Respiratory Syndrome* / metabolism
Porcine respiratory and reproductive syndrome virus* / metabolism
RNA, Long Noncoding* / genetics
RNA, Long Noncoding* / metabolism
Swine
Transcription Factors / genetics
Virus Replication

Substances

3' Untranslated Regions
DNA-Binding Proteins
Interleukin-6
MicroRNAs
NF-kappa B
RNA, Long Noncoding
Transcription Factors

Grants and funding

This work was supported by Open Funds of the State Key Laboratory of Agricultural Microbiology (AMLKF202104), Fundamental Research Funds for the Central Universities (2662020DKPY003) and National Natural Science Foundation of China (31201764) for M.L. and by National Key Research and Development Program of China (2021YFD1301201), National Natural Science Foundation of China (31172179), Natural Science Foundation of Hubei Province (2015CFA101), Agricultural Science and Technology Innovation Action of Hubei Province (2018skjcx01), Agricultural Science Innovation Foundation of Hubei Province (2021-620-000-001-030), Animal Husbandry Major Technology Collaborative Extension Project of Hubei Province and Research Project of Huazhong Agricultural University for D.X. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. None of the authors received a salary from their patrons.