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Design of antiviral AGO2-dependent short hairpin RNAs

Virologica Sinica 2024年第4期39卷 645-654页

作者： Yuanyuan Bie Jieling Zhang Jiyao Chen Yumin Zhang Muhan Huang Leike Zhang Xi Zhou Yang Qiu Key Laboratory of virology and Biosafety Wuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China State Key Laboratory of virology Wuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China University of Chinese Academy of Sciences Beijing 100049China School of Life Sciences Division of Life Sciences and MedicineUniversity of Science and Technology of ChinaHefei 230027China

The increasing emergence and re-emergence of RNA virus outbreaks underlines the urgent need to develop effective *** interference(RNAi)is a sequence-specific gene silencing mechanism that is triggered by small interfering RNAs(siRNAs)or short hairpin RNAs(shRNAs),which exhibits significant promise for antiviral ***2-dependent shRNA(agshRNA)generates a single-stranded guide RNA and presents significant advantages over traditional siRNA and *** this study,we applied a logistic regression algorithm to a previously published chemically siRNA efficacy dataset and built a machine learning-based model with high predictive *** this model,we designed siRNA sequences targeting diverse RNA viruses,including human enterovirus A71(EV71),Zika virus(ZIKV),dengue virus 2(DENV2),mouse hepatitis virus(MHV)and severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),and transformed them into *** validated the performance of our agshRNA design by evaluating antiviral efficacies of agshRNAs in cells infected with different *** the agshRNA targeting EV71 as an example,we showed that the anti-EV71 effect of agshRNA was more potent compared with the corresponding siRNA and ***,the antiviral effect of agshRNA is dependent on AGO2-processed guide RNA,which can load into the RNA-induced silencing complex(RISC).We also confirmed the antiviral effect of agshRNA in ***,this work develops a novel antiviral strategy that combines machine learning-based algorithm with agshRNA design to custom design antiviral agshRNAs with high efficiency.

关键词： AgshRNA Antiviral strategy Machine learning model

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Mutations in the SARS-CoV-2 spike receptor binding domain and their delicate balance between ACE2 affinity and antibody evasion

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Protein & Cell 2024年第6期15卷 403-418页

作者： Song Xue Yuru Han Fan Wu Qiao Wang Key Laboratory of medical Molecular virology (MOE/NHC/CAMS) Shanghai Institute of Infectious Disease and BiosecurityShanghai Frontiers Science Center of Pathogenic Microorganisms and InfectionSchool of Basic Medical SciencesShanghai Medical CollegeFudan UniversityShanghai 200032China

Intensive selection pressure constrains the evolutionary trajectory of SARS-CoV-2 genomes and results in various novel variants with distinct mutation *** mutations,particularly those within the receptor binding domain(RBD)of SARS-CoV-2 spike(S)protein,lead to the functional alteration in both receptor engagement and monoclonal antibody(mAb)***,we review the data of the RBD point mutations possessed by major SARS-CoV-2 variants and discuss their individual effects on ACE2 affinity and immune *** single amino acid substitutions within RBD epitopes crucial for the antibody evasion capacity may conversely weaken ACE2 binding ***,this weakened effect could be largely compensated by specific epistatic mutations,such as N501Y,thus maintaining the overall ACE2 affinity for the spike protein of all major *** predominant direction of SARS-CoV-2 evolution lies neither in promoting ACE2 affinity nor evading mAb neutralization but in maintaining a delicate balance between these two ***,this review interprets how RBD mutations efficiently resist antibody neutralization and meanwhile how the affinity between ACE2 and spike protein is maintained,emphasizing the significance of comprehensive assessment of spike mutations.

关键词： ACE2 affinity maintained

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Single-cell RNA sequencing to understand host-virus interactions

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Virologica Sinica 2024年第1期39卷 1-8页

作者： Jia-Tong Chang Li-Bo Liu Pei-Gang Wang Jing An Department of microbiology School of Basic Medical SciencesCapital Medical UniversityBeijing 100069China

Single-cell RNA sequencing(scRNA-seq)has allowed for the profiling of host and virus transcripts and host-virus interactions at single-cell *** review summarizes the existing scRNA-seq technologies together with their strengths and *** applications of scRNA-seq in various virological studies are discussed in depth,which broaden the understanding of the immune atlas,host-virus interactions,and immune ***-seq can be widely used for virology in the near future to better understand the pathogenic mechanisms and discover more effective therapeutic strategies.

关键词： Single-cell RNA sequencing(scRNA-seq) Host-virus interaction COVID-19 Flavivirus

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Leafhopper salivary carboxylesterase suppresses JA-Ile synthesis to facilitate initial arbovirus transmission in rice phloem

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Plant Communications 2024年第9期5卷 116-135页

作者： Yunhua Chi Hongxiang Zhang Siyu Chen Yu Cheng Xiaofeng Zhang Dongsheng Jia Qian Chen Hongyan Chen Taiyun Wei Vector-borne Virus Research Center Fujian Province Key Laboratory of Plant VirologyInstitute of Plant VirologyFujian Agriculture and Forestry UniversityFuzhouFujian 350002China

Plant jasmonoyl-L-isoleucine(JA-Ile)is a major defense signal against insect feeding,but whether or how insect salivary effectors suppress JA-Ile synthesis and thus facilitate viral transmission in the plant phloem remains *** carboxylesterases(CarEs)are the third major family of detoxification ***,we identify a new leafhopper CarE,CarE10,that is specifically expressed in salivary glands and is secreted into the rice phloem as a saliva *** CarE10 directly binds to rice jasmonate resistant 1(JAR1)and promotes its degradation by the proteasome ***,the direct association of CarE10 with JAR1 clearly impairs JAR1 enzyme activity for conversion of JA to JA-Ile in an in vitro JAIle synthesis system.A devastating rice reovirus activates and promotes the co-secretion of virions and CarE10 via virus-induced vesicles into the saliva-storing salivary cavities of the leafhopper vector and ultimately into the rice phloem to establish initial ***,a virus-mediated increase in CarE10 secretion or overexpression of CarE10 in transgenic rice plants causes reduced levels of JAR1 and thus suppresses JA-Ile synthesis,promoting host attractiveness to insect vectors and facilitating initial viral *** findings provide insight into how the insect salivary protein CarE10 suppresses host JA-Ile synthesis to promote initial virus transmission in the rice phloem.

关键词： leafhopper salivary protein CarE10 JAR1 degradation JA-Ile synthesis rice phloem initial viral transmission

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Crucial role played by CK8^(+)cells in mediating alveolar injury remodeling for patients with COVID-19

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Virologica Sinica 2024年第3期39卷 390-402页

作者： Yufeng Li Hengrui Hu Jia Liu Longda Ma Xi Wang Liang Liu Qian Liu Liang Ren Jiang Li Fei Deng Zhihong Hu Yiwu Zhou Manli Wang State Key Laboratory of virology Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of SciencesWuhan430071China Department of Forensic Medicine Tongji Medical College of Huazhong University of Science and TechnologyWuhan430010China University of the Chinese Academy of Sciences Beijing100049China

The high risk of SARS-CoV-2 infection and reinfection and the occurrence of post-acute pulmonary sequelae have highlighted the importance of understanding the mechanism underlying lung repair after *** address this concern,comparative and systematic analyses of SARS-CoV-2 infection in COVID-19 patients and animals were *** the lungs of nine patients who died of COVID-19 and one recovered from COVID-19 but died of unrelated disease in early 2020,damage-related transient progenitor(DATP)cells expressing CK8 marker proliferated *** CK8^(+)DATP cells were derived from bronchial CK5^(+)basal ***,they showed different cell fate toward differentiation into type I alveolar cells in the deceased and convalescent patients,*** using a self-limiting hamster infection model mimicking the dynamic process of lung injury remodeling in mild COVID-19 patients,the accumulation and regression of CK8t cell marker were found to be closely associated with the disease ***,we examined the autopsied lungs of two patients who died of infection by the recent Omicron variant and found that they only exhibited mild pathological injury with no CK8^(+)cell *** results indicate a clear pulmonary cell remodeling route and suggest that CK8^(+)DATP cells play a primary role in mediating alveolar remodeling,highlighting their potential applications as diagnostic markers and therapeutic targets.

关键词： SARS-CoV-2 CK8t DATP cells Cell proliferation Lung injury-remodeling Autopsy Hamster infection model

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AIMP2 restricts EV71 replication by recruiting SMURF2 to promote the degradation of 3D polymerase

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Virologica Sinica 2024年第4期39卷 632-644页

作者： Junrui Ren Lei Yu Qiuhan Zhang Pengyu Ren Yumeng Cai Xueyun Wang Ke Lan Shuwen Wu State Key Laboratory of virology College of Life SciencesWuhan UniversityWuhan430072China medical Research institute Wuhan UniversityWuhan430072China Taikang Center for Life and medical Sciences Wuhan UniversityWuhan430072China

Hand,foot and mouth disease(HFMD),mainly caused by enterovirus 71(EV71),has frequently occurred in the Asia-Pacific region,posing a significant threat to the health of infants and young ***,research on the infection mechanism and pathogenicity of enteroviruses is increasingly becoming *** 3D polymerase,as the most critical RNA-dependent RNA polymerase(RdRp)for EV71 replication,is widely targeted to inhibit EV71 *** this study,we identified a novel host protein,AIMP2,capable of binding to 3D polymerase and inhibiting EV71 *** investigations revealed that AIMP2 recruits the E3 ligase SMURF2,which mediates the polyubiquitination and degradation of 3D ***,the antiviral effect of AIMP2 extended to the CVA16 and CVB1 *** research has uncovered the dynamic regulatory function of AIMP2 during EV71 infection,revealing a novel antiviral mechanism and providing new insights for the development of antienteroviral therapeutic strategies.

关键词： AIMP2 Enterovirus 71(EV71) 3D polymerase E3 ligase SMURF2 Ubiquitination

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Screening of Rodents from Rural Communities of Lagos and Ogun State Nigeria of Lassa Virus RNA

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Open Journal of medical microbiology 2024年第2期14卷 120-130页

作者： Fatai Sarafa Emmanuel Tochukwu Onyeje Joseph Ojonugwa Shaibu Olumuyiwa Babalola Salu Rosemary Ajuma Audu Sunday Aremu Omilabu Department of medical microbiology and Parasitology College of Medicine University of Lagos Lagos Nigeria Centre for Human virology and Genomics Research Microbiology Department Nigerian Institute of Medical Research Lagos Nigeria Centre for Human and Zoonotic virology Central Research Laboratory and Department of Medical Microbiology and Parasitology College of Medicine University of Lagos Lagos Nigeria

Background: In recent times, there has been an increase in the number of Lassa fever cases resulting from the several episodes of Lassa fever epidemics ravaging Nigeria and other West African countries. The presence of Lassa virus in rodents other than the major reservoir (Mastomys natalensis) has been a public health concern as to the actual burden of the disease. It is therefore of a public health necessity to explore the LASV RNA habouring potential of several species of rodents in endemic as well as non-endemic areas for proper prevention of emergence of outbreaks in non-endemic areas. Objectives: The aim of this study was to detect the presence of LASV RNA in different species of rodents in Ikorodu, Lagos state and Abeokuta, Ogun state. Methods: A total of ninety one (91) rodents were captured from Ikorodu, Lagos State (61 rodents) and Abeokuta, Ogun State (30 rodents), euthanized, bled, and plasma obtained for the detection of LASV RNA by Reverse Transcriptase Polymerase Chain Reaction. Results: A total of 91 rodents consisting of 77 Rattus rattus and 14 Crocidura spp. The S segment of LASV RNA was not in any of the 91 rodents’ plasma samples. Conclusion: The rodents captured within the rural communities of Ikorodu, Lagos State and Abeokuta, Ogun State were found not to habour the LASV RNA. This study is limited by the relatively small sample size. Similar studies should be encouraged both in endemic and non-endemic areas in order to understand the actual burden of Lassa fever as well as put into check future epidemics.

关键词： Lassa Virus Lassa Fever Mastomys natalensis Transmission Lagos State Ogun State

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Identification of residues in Lassa virus glycoprotein 1 involved in receptor switch

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Virologica Sinica 2024年第4期39卷 600-608页

作者： Jiao Guo Yi Wan Yang Liu Xiaoying Jia Siqi Dong Gengfu Xiao Wei Wang State Key Laboratory of virology Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of SciencesWuhan 430071China University of the Chinese Academy of Sciences Beijing 100049China The Xi'an Key Laboratory of Pathogenic Microorganism and Tumor Immunity School of Basic MedicineXi'an Medical UniversityXi'an 710021China

Lassa virus(LASV)is an enveloped,negative-sense RNA virus that causes Lassa hemorrhagic *** entry of LASV requires the viral glycoprotein 1(GP1)to undergo a receptor switch from its primary receptor alpha-dystroglycan(α-DG)to its endosomal receptor lysosome-associated membrane protein 1(LAMP1).A conserved histidine triad in LASV GP1 has been reported to be responsible for receptor *** test the hypothesis that other non-conserved residues also contribute to receptor switch,we constructed a series of mutant LASV GP1 proteins and tested them for binding to *** residues,L84,K88,L107,and H170,were identified as critical for receptor *** any of the four residues with the corresponding lymphocytic choriomeningitis virus(LCMV)residue(L84 N,K88E,L10F,and H170S)reduced the binding affinity of LASV GP1 for ***,all mutations caused decreases in glycoprotein precursor(GPC)-mediated membrane fusion at both pH 4.5 and *** infectivity of pseudotyped viruses bearing either GPCL84N or GPCK88E decreased sharply in multiple cell types,while L107F and H170S had only mild effects on *** biolayer light interferometry assay,we found that all four mutants had decreased binding affinity to LAMP1,in the order of binding affinity being L84 N>L107F>K88E>*** four amino acid loci identified for the first time in this study have important reference significance for the in-depth investigation of the mechanism of receptor switching and immune escape of LASV occurrence and the development of reserve anti-LASV infection drugs.

关键词： Lassa virus(LASV) Lysosome-associated membrane protein 1 (LAMP1) Glycoprotein Receptor switch Membrane fusion

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Minimizing the ratio of ionizable lipid in lipid nanoparticles for in vivo base editing

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National Science Review 2024年第6期11卷 285-293页

作者： Qiubing Chen Xuebin Wang Yizhou Zhang Ming Tian Junyi Duan Ying Zhang Hao Yin Departments of Urology and Laboratory Medicine Frontier Science Center for Immunology and Metabolism Medical Research InstituteZhongnan Hospital of Wuhan UniversityWuhan University State Key Laboratory of virology Tai Kang Center for Life and Medical Sciences Tai Kang Medical SchoolWuhan University Department of Rheumatology and Immunology Zhongnan Hospital of Wuhan UniversityWuhan University Wuhan Research Center for Infectious Diseases and Cancer Chinese Academy of Medical Sciences RNA institute Wuhan University

Lipid nanoparticles(LNPs) have gained clinical approval as carriers for both siRNA and mRNA. Among the crucial components of LNPs, ionizable lipids play a pivotal role in determining the efficiency of RNA delivery. In this study, we synthesized a series of ionizable lipids, denoted as HTO, with a higher count of hydroxyl groups compared to SM-102. Remarkably, LNPs based on HTO12 lipid demonstrated comparable mRNA delivery efficiency and biosafety to those based on SM-102. However, the former reduced the ratio of ionizable lipid/total lipids to mRNA in LNPs by 2.5 times compared to SM-102. The HTO12 LNP efficiently encapsulated adenine base editor mRNA and sgRNA targeting Pcsk9, leading to substantial gene editing within the liver of mice and effective reduction of the target protein. Our study underscores that ionizable lipids with multiple hydroxyl groups may facilitate an improved lipid-to-mRNA ratio to minimize the dosage of ionizable lipids for in vivo delivery.

关键词： ionizable lipid lipid nanoparticle mRNA delivery base editor delivery Pcsk9

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C1QTNF5 is a novel attachment factor that facilitates the entry of influenza A virus

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Virologica Sinica 2024年第2期39卷 277-289页

作者： Lei Yu Xinjin Liu Xiaoqin Wei Junrui Ren Xueyun Wang Shuwen Wu Ke Lan State Key Laboratory of virology Medical Research InstituteCollege of Life SciencesWuhan UniversityWuhan430072China Frontier Science Center for Immunology and Metabolism Wuhan UniversityWuhan430072China Taikang Center for Life and medical Sciences Wuhan UniversityWuhan430072China

Influenza A virus(IAV)binds sialic acid receptors on the cell surface to enter the host cells,which is the key step in initiating infection,transmission and *** the factors that contribute to the highly efficient entry of IAV into human cells will help elucidate the mechanism of viral entry and pathogenicity,and provide new targets for *** the present study,we reported a novel membrane protein,C1QTNF5,which binds to the hemagglutinin protein of IAV and promotes IAV infection in vitro and in *** found that the HA1 region of IAV hemagglutinin is critical for the interaction with C1QTNF5 protein,and C1QTNF5 interacts with hemagglutinin mainly through its N-terminus(1–103 aa).In addition,we further demonstrated that overexpression of C1QTNF5 promotes IAV entry,while blocking the interaction between C1QTNF5 and IAV hemagglutinin greatly inhibits viral ***,C1QTNF5 does not function as a receptor to mediate IAV infection in sialic acid-deficient CHO-Lec2 cells,but promotes IAV to attach to these cells,suggesting that C1QTNF5 is an important attachment factor for *** work reveals C1QTNF5 as a novel IAV attachment factor and provides a new perspective for antiviral strategies.

关键词： C1QTNF5 Influenza A virus Viral entry Hemagglutinin Attachment factor

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