miR-4454通过靶向LRPAP1激活Ⅰ型干扰素信号通路抑制寨卡病毒复制*

doi:10.3969/j.issn.1671-2587.2025.03.009

临床输血与检验 ›› 2025, Vol. 27 ›› Issue (3): 329-339.DOI: 10.3969/j.issn.1671-2587.2025.03.009

miR-4454通过靶向LRPAP1激活Ⅰ型干扰素信号通路抑制寨卡病毒复制^*

韦芊芊, 陈利民, 李玉佳, 李世林

中国医学科学院北京协和医学院输血研究所, 四川成都 610052

收稿日期:2025-02-28 发布日期:2025-06-23
通讯作者: 李世林,主要从事输血传播病原体的致病机制研究,（E-mail）shilin-li@hotmail.com。共同通信作者李玉佳,主要从事输血传播病原体研究,（E-mail）lily83630@163.com。
作者简介:韦芊芊,主要从事输血传播病原体的致病机制研究,（E-mail）916157386@qq.com。
基金资助:
*本课题受四川省自然科学基金（面上）项目（No.2025ZNSFSC0678）资助

MiR-4454 Inhibits Zika Virus Replication by Targeting LRPAP1 and Activating the Type Ⅰ Interferon Signaling Pathway

WEI Qianqian, CHEN Limin, LI Yujia, LI Shilin

Institute of Blood Transfusion, Chinese Academy of Medical Sciences Peking Union Medical College, Chengdu 610052

Received:2025-02-28 Published:2025-06-23

摘要/Abstract

摘要： 目的本研究探讨miR-4454对寨卡病毒（ZIKV）复制的调节作用及其潜在分子机制。方法将miR-4454的模拟物（mimic）或inhibitor转染至细胞后,用ZIKV感染,qRT-PCR或western blot分别检测ZIKV NS5 mRNA及NS1蛋白的表达。使用生物信息学软件对miR-4454可能的靶基因进行预测,并验证miR-4454下游靶基因LRPAP1-低密度脂蛋白受体相关蛋白关联蛋白1（low-density lipoprotein receptor-related protein associated protein 1, LRPAP1）。将LRPAP1质粒或si-LRPAP1转染至细胞后,再用ZIKV感染,qRT-PCR检测ZIKV NS5 mRNA、干扰素刺激基因的表达,Western blot 检测ZIKV NS1蛋白、IFNAR1和STAT1磷酸化水平,探究LRPAP1表达改变对ZIKV复制和Ⅰ型干扰素信号通路的影响。结果在细胞中高表达miR-4454抑制ZIKV NS5 mRNA的复制或NS1蛋白的表达。预测得到miR-4454潜在靶基因LRPAP1,其与病毒感染和Ⅰ型干扰素信号通路有关,双荧光素酶报告基因实验证实LRPAP1是miR-4454直接作用的靶基因之一。过表达LRPAP1促进ZIKV复制和抑制Ⅰ型干扰素信号通路,而敲低LRPAP1抑制ZIKV复制以及激活Ⅰ型干扰素信号通路。结论 miR-4454通过固有免疫通路调控ZIKV感染,揭示了其作为潜在靶点的可能性,为相关治疗策略的开发提供了新思路。

关键词: 寨卡病毒, 非编码RNA, miR-4454, Ⅰ型干扰素信号通路

Abstract: Objective This study aims to investigate the specific regulatory effect of miR-4454 on ZIKV replication and reveal its potential molecular mechanisms, providing new ideas and targets for the treatment of ZIKV infection. Methods Cells were transfected with miR-4454 mimics or inhibitors and subsequently infected with ZIKV. The expression of ZIKV NS5 mRNA and NS1 protein was measured using qRT-PCR and Western blot analyses. Bioinformatics tools were employed to predict potential target genes of miR-4454, focusing on the validation of LRPAP1 (low-density lipoprotein receptor-related protein associated protein 1) as a downstream target. Following the transfection of LRPAP1 plasmid or si-LRPAP1 into cells, ZIKV infection was performed, and qRT-PCR was used to detect ZIKV NS5 mRNA and interferon-stimulated gene expression, while Western blotting assessed the levels of ZIKV NS1 protein, IFNAR1, and phosphorylated STAT1, to explore the effect of LRPAP1 expression changes on ZIKV replication and the Type Ⅰ interferon signaling pathway. Results Overexpression of miR-4454 in cells inhibited the replication of ZIKV NS5 mRNA and expression of NS1 protein. Bioinformatics prediction identified LRPAP1 as a potential target of miR-4454, which is related to viral infection and the Type Ⅰ interferon signaling pathway. Dual-luciferase reporter assays confirmed that LRPAP1 is a direct target of miR-4454. Overexpression of LRPAP1 promoted ZIKV replication and suppressed the Type Ⅰ interferon signaling pathway, while knockdown of LRPAP1 inhibited ZIKV replication and activated this signaling pathway. Conclusion miR-4454 modulates ZIKV infection through the intrinsic immune pathway, suggesting its potential as a therapeutic target and providing new insights for the development of related treatment strategies.

Key words: Zika virus, Non-coding RNA, MiR-4454, Type Ⅰ interferon signaling pathway

中图分类号:

R373.9

韦芊芊, 陈利民, 李玉佳, 李世林. miR-4454通过靶向LRPAP1激活Ⅰ型干扰素信号通路抑制寨卡病毒复制^*[J]. 临床输血与检验, 2025, 27(3): 329-339.

WEI Qianqian, CHEN Limin, LI Yujia, LI Shilin. MiR-4454 Inhibits Zika Virus Replication by Targeting LRPAP1 and Activating the Type Ⅰ Interferon Signaling Pathway[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2025, 27(3): 329-339.

参考文献

[1] DICK G A,KITCHEN S F,HADDOW A J.Zika virus. I. isolations and serological specificity[J]. Trans R Soc Trop Med Hyg,1952,46(5):509-520.
[2] GRARD G,CARON M,MOMBO I M,et al.Zika virus in Gabon (central Africa)—2007:a new threat from Aedes albopictus?[J]. PLoS Negl Trop Dis,2014,8(2):e2681.
[3] CAO-LORMEAU V M,ROCHE C,TEISSIER A, et al. Zika virus,French Polynesia,south Pacific,2013[J]. Emerg Infect Dis,2014,20(6):1085-1086.
[4] BESNARD M,LASTERE S,TEISSIER A,et al.Evidence of perinatal transmission of zika virus,French Polynesia,December 2013 and February 2014[J]. Euro Surveill,2014,19(13):20751.
[5] MUSSO D,BOSSIN H,MALLET H P,et al.Zika virus in French Polynesia 2013-14:anatomy of a completed outbreak[J]. Lancet Infect Dis,2018,18(5):e172-e182.
[6] WHO.WHO Director-General summarizes the outcome of the Emergency Committee regarding clusters of microcephaly and Guillain-Barré syndrome[EB/OL].[2025-01-20]. https://www.who.int/news/item/01-02-2016-who-director-general-summarizes-the-outcome-of-the-emergency-committee-regarding-clusters-of-microcephaly-and-guillain-barré-syndrome.
[7] MUSSO D,KO A I,BAUD D.Zika virus infection-after the pandemic[J]. N Engl J Med,2019,381(15):1444-1457.
[8] FERNANDEZ-GARCIA M D,MAZZON M, JACOBS M,et al. Pathogenesis of flavivirus infections:using and abusing the host cell[J]. Cell Host Microbe,2009, 5(4):318-328.
[9] KHARWADKAR S,HERATH N.Clinical manifestations of dengue,zika and chikungunya in the Pacific Islands:a systematic review and meta-analysis[J]. Rev Med Virol, 2024,34(2):e2521.
[10] FENG Y Q.Recent advances in the study of zika virus structure,drug targets,and inhibitors[J]. Front Pharmacol,2024,15:1418516.
[11] GARLEY M,NOWAK K,JABŁOŃSKA E. Neutrophil microRNAs[J]. Biol Rev Camb Philos Soc,2024,99(3): 864-877.
[12] LI H Y,WANG X F,MU H B,et al.miR-484 contributes to diminished ovarian reserve by regulating granulosa cell function via YAP1-mediated mitochondrial function and apoptosis[J]. Int J Biol Sci,2022,18(3):1008-1021.
[13] LU S,ZHU N,GUO W W,et al.RNA-seq revealed a circular RNA-microRNA-mRNA regulatory network in hantaan virus infection[J]. Front Cell Infect Microbiol, 2020,10:97.
[14] BAHOJB MAHDAVI S Z,JEBELLI A,AGHBASH P S, et al. A comprehensive overview on the crosstalk between microRNAs and viral pathogenesis and infection[J]. Med Res Rev,2025,45(2):349-425.
[15] AZOUZ F,ARORA K,KRAUSE K,et al.Integrated microRNA and mRNA profiling in zika virus-infected neurons[J]. Viruses,2019,11(2):162.
[16] YE H Y,KANG L,YAN X P,et al.miR-103a-3p promotes zika virus replication by targeting OTU deubiquitinase 4 to activate p38 mitogen-activated protein kinase signaling pathway[J]. Front Microbiol,2022,13:862580.
[17] BU G,SCHWARTZ A L.RAP,a novel type of ER chaperone[J]. Trends Cell Biol,1998,8(7):272-276.
[18] WILLNOW T E,ROHLMANN A,HORTON J,et al.RAP,a specialized chaperone,prevents ligand-induced ER retention and degradation of LDL receptor-related endocytic receptors[J]. EMBO J,1996,15(11):2632-2639.
[19] LI H C,WANG X,WANG Y R,et al.Secreted LRPAP1 binds and triggers IFNAR1 degradation to facilitate virus evasion from cellular innate immunity[J]. Signal Transduct Target Ther,2023,8(1):374.
[20] SHEREEN M A,BASHIR N,SU R,et al.Zika virus dysregulates the expression of astrocytic genes involved in neurodevelopment[J]. PLoS Negl Trop Dis,2021, 15(4):e0009362.
[21] WU W J,WANG C,XIA C L,et al.microRNA let-7 suppresses influenza A virus infection by targeting RPS16 and enhancing type Ⅰ interferon response[J]. Front Cell Infect Microbiol,2022,12:904775.
[22] JAFARZADEH A,NASERI A,SHOJAIE L,et al.microRNA-155 and antiviral immune responses[J]. Int Immunopharmacol,2021,101(Pt A):108188.
[23] WANG H,HU H,LUO Z Z,et al. miR-4454 up-regulated by HPV16 E6/E7 promotes invasion and migration by targeting ABHD2/NUDT21 in cervical cancer[J]. Biosci Rep,2020,40(9):BSR20200796.
[24] GUO X K,ZHANG Q,GAO L,et al.Increasing expression of microRNA 181 inhibits porcine reproductive and respiratory syndrome virus replication and has implications for controlling virus infection[J]. J Virol,2013,87(2):1159-1171.
[25] SCHNEIDER W M,CHEVILLOTTE M D,RICE C M.Interferon-stimulated genes:a complex web of host defenses[J]. Annu Rev Immunol,2014,32:513-545.
[26] SCHOGGINS J W,WILSON S J,PANIS M,et al.A diverse range of gene products are effectors of the typeⅠ interferon antiviral response[J]. Nature,2011, 472: 481-485.
[27] PAREEK S,ROY S,KUMARI B,et al.miR-155 induction in microglial cells suppresses Japanese encephalitis virus replication and negatively modulates innate immune responses[J]. J Neuroinflammation,2014,11:97.
[28] THOUNAOJAM M C,KAUSHIK D K,KUNDU K, et al.microRNA-29b modulates Japanese encephalitis virus-induced microglia activation by targeting tumor necrosis factor alpha-induced protein 3[J]. J Neurochem, 2014,129(1):143-154.
[29] BUGGELE W A,HORVATH C M. microRNA profiling of Sendai virus-infected A549 cells identifies miR-203 as an interferon-inducible regulator of IFIT1/ISG56[J]. J Virol,2013,87(16):9260-9270.
[30] FENG L X,FENG Z A,HU J,et al.Identification of hsa-miR-619-5p and hsa-miR-4454 in plasma-derived exosomes as a potential biomarker for lung adenocarcinoma[J]. Front Genet,2023,14:1138230.
[31] LIN H M,ZHANG R,WU W R,et al.miR-4454 promotes hepatic carcinoma progression by targeting Vps4A and Rab27A[J]. Oxid Med Cell Longev,2021,2021:9230435.
[32] ZHAO L N,YUAN H F,WANG Y F,et al.HBV confers innate immune evasion through triggering HAT1/acetylation of H4K5/H4K12/miR-181a-5p or KPNA2/cGAS-STING/IFN-I signaling[J]. J Med Virol,2023, 95(7): e28966.
[33] PAN Q,XIE Y,ZHANG Y,et al.EGFR core fucosylation,induced by hepatitis C virus,promotes TRIM40-mediated-RIG-I ubiquitination and suppresses interferon-I antiviral defenses[J]. Nat Commun,2024, 15(1):652.
[34] MORRISON J,LAURENT-ROLLE M,MAESTRE A M,et al.Dengue virus co-opts UBR4 to degrade STAT2 and antagonize type Ⅰ interferon signaling[J]. PLoS Pathog,2013,9(3):e1003265.
[35] SINGH P K,GUEST J M,KANWAR M,et al.Zika virus infects cells lining the blood-retinal barrier and causes chorioretinal atrophy in mouse eyes[J]. JCI Insight, 2017,2(4):e92340.
[36] SINGH P K,SINGH S,FARR D,et al.Interferon-stimulated gene 15 (ISG15) restricts Zika virus replication in primary human corneal epithelial cells[J]. Ocul Surf,2019,17(3):551-559.
[37] ESPADA C E,DA ROCHA E L,RICCIARDI-JORGE T, et al. ISG15/USP18/STAT2 is a molecular hub regulating IFN I-mediated control of Dengue and Zika virus replication[J]. Front Immunol,2024,15:1331731.
[38] DAI J F,PAN W,WANG P H.ISG15 facilitates cellular antiviral response to dengue and west Nile virus infection in vitro[J]. Virol J,2011,8:468.
[39] LIU H C,YAN L,LI X K,et al.microRNA expression in osteoarthritis:a meta-analysis[J]. Clin Exp Med,2023, 23(7):3737-3749.
[40] ZHOU C,ZHAO X,DUAN S W.The role of miR-543 in human cancerous and noncancerous diseases[J]. J Cell Physiol,2021,236(1):15-26.
[41] BAUER A N,MAJUMDAR N,WILLIAMS F,et al.microRNAs:small but key players in viral infections and immune responses to viral pathogens[J]. Biology (Basel), 2023,12(10):1334.

miR-4454通过靶向LRPAP1激活Ⅰ型干扰素信号通路抑制寨卡病毒复制^*

MiR-4454 Inhibits Zika Virus Replication by Targeting LRPAP1 and Activating the Type Ⅰ Interferon Signaling Pathway

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