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Overexpression of wild-type HRAS drives non-alcoholic steatohepatitis to hepatocellular carcinoma in mice

Zoological Research 2024年第3期45卷 551-566页

作者： Chen Ling Su-Su Liu Yu-Ya Wang Gui-Tao Huo Yan-Wei Yang Nan Xu Hong Wang Yong Wu Yu-Fa Miao Rui Fu Yu-Wei Zhao Chang-Fa Fan College of Life Sciences Northwest UniversityProvincial Key Laboratory of Biotechnology of Shaanxi ProvinceXi’anShaanxi 710069China division of Animal Model Research Institute for Laboratory Animal ResourcesNational Institutes for Food and Drug Control(NIFDC)Beijing 102629China National Center for Safety Evaluation of Drugs National Institutes for Food and Drug Control(NIFDC)Beijing 100176China division of hiv/aids and sexually transmitted virus vaccines Institute for Biological Products ControlNational Institutes for Food and Drug Control(NIFDC)Beijing 102629China division of Laboratory Animal Monitoring Institute for Laboratory Animal ResourcesNational Institutes for Food and Drug Control(NIFDC)Beijing 102629China

Hepatocellular carcinoma(HCC),a prevalent solid carcinoma of significant concern,is an aggressive and often fatal disease with increasing global incidence rates and poor therapeutic *** etiology and pathological progression of non-alcoholic steatohepatitis(NASH)-related HCC is multifactorial and ***,no single animal model can accurately mimic the full NASH-related HCC pathological progression,posing considerable challenges to transition and mechanistic ***,a novel conditional inducible wild-type human HRAS overexpressed mouse model(HRAS-HCC)was established,demonstrating 100%morbidity and mortality within approximately one month under normal dietary and lifestyle *** symptoms of HCC such as ascites,thrombus,internal hemorrhage,jaundice,and lung metastasis were successfully replicated in ***-depth pathological features of NASH-related HCC were demonstrated by pathological staining,biochemical analyses,and typical marker gene *** murine anti-PD-1 and sorafenib treatment effectively prolonged mouse survival,further confirming the accuracy and reliability of the *** on protein-protein interaction(PPI)network and RNA sequencing analyses,we speculated that overexpression of HRAS may initiate the THBS1-COL4A3 axis to induce NASH with severe fibrosis,with subsequent progression to ***,our study successfully duplicated natural sequential progression in a single murine model over a very short period,providing an accurate and reliable preclinical tool for therapeutic evaluations targeting the NASH to HCC continuum.

关键词： HRAS THBS1 HCC driver factor NASH Fibrosis Cirrhosis HCC Treatment evaluation

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Screening and Identification of Marburg virus Entry Inhibitors Using Approved Drugs

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Virologica Sinica 2020年第2期35卷 235-239页

作者： Li Zhang Shan Lei Hui Xie Qianqian Li Shuo Liu Qiang Liu Weijin Huang Xinyue Xiao Youchun Wang division of hiv/aids and sexually-transmitted virus vaccines National Institutes for Food and Drug ControlBeijing 102629China Institute for Reference Standards and Standardization National Institutes for Food and Drug ControlBeijing 102629China

Dear Editor,Marburg virus(MARV)belongs to the Filoviridae family,along with the related Ebola virus(EBOV).Although MARV is less renowned than EBOV,it causes an equally devastating disease,with clinical symptoms similar to EBOV infection and a remarkably high mortality *** date,about 600 MARV cases have been reported globally,with an average mortality rate of around 80%(Brauburger et al.2012).Due to the severity of disease caused by MARV,as well as its threat to global public health,MARV is classified as a risk group 4 pathogen by WHO,necessitating containment measures equivalent to biosafety level 4(BSL-4).

关键词： mortality al. clinical

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Monitoring Neutralization Property Change of Evolving Hantaan and Seoul viruses with a Novel Pseudovirus-Based Assay

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Virologica Sinica 2021年第1期36卷 104-112页

作者： Tingting Ning Ling Wang Shuo Liu Jian Ma Jianhui Nie Weijin Huang Xuguang Li Yuhua Li Youchun Wang division of hiv/aids and sexually transmitted virus vaccines National Institutes for Food and Drug Control(NIFDC)Beijing 102629China division of Arboviruses Vaccine National Institutes for Food and Drug Control(NIFDC)Beijing 102629China Centre for Biologics Evaluation Biologics and Genetic Therapies DirectorateHealth CanadaOttawaON K2J 4X9Canada

The Hantaan virus(HTNV)and Seoul virus(SEOV)mutants have accumulated over *** is important to determine whether their neutralizing epitopes have evolved,thereby making the current vaccine ***,it is impossible to determine by using traditional plaque reduction neutralization test(PRNT),because it requires large numbers of live mutant ***-based neutralization assays(PBNA)were developed by employing vesicular stomatitis virus(VSV)backbone incorporated with HTNV or SEOV glycoproteins(VSVDG*-HTNVG or VSVDG*-SEOVG).56 and 51 single amino acid substitutions of glycoprotein(GP)in HTNV and SEOV were selected and introduced into the reference *** the mutant pseudoviruses were generated and tested by *** PBNA results were highly correlated with PRNT ones with R2 being 0.91 for VSVDG*-HTNVG and 0.82 for VSVDG*-SEOVG.53 HTNV mutant pseudoviruses and 46 SEOV mutants were successfully ***,by using PBNA,we found that HTNV or SEOV immunized antisera could neutralize all the corresponding 53 HTNV mutants or the 46 SEOV mutants *** novel PBNA enables us to closely monitor the effectiveness of vaccines against large numbers of evolving HTNV and *** the current vaccine remains to be effective for the naturally occurring mutants.

关键词： Hemorrhagic fever with renal syndrome(HFRS) Hantaan virus(HTNV) Seoul virus(SEOV) Pseudovirus-based neutralization assay(PBNA) Amino acid substitution Vaccine

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Three amino acid residues in the envelope of human immunodeficiency virus type 1 CRF07_BC regulate viral neutralization susceptibility to the human monoclonal neutralizing antibody IgG1b12

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Virologica Sinica 2014年第5期29卷 299-307页

作者： Jianhui Nie Juan Zhao Qingqing Chen Weijin Huang Youchun Wang College of Life Science Jilin University Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products Division of HIV/AIDS and sex-transmitted virus vaccines National Institutes for Food and Drug Control (NIFDC)

The CD4 binding site(CD4bs) of envelope glycoprotein(Env) is an important conserved target for anti-human immunodeficiency virus type 1(hiv-1) neutralizing antibodies. Neutralizing monoclonal antibodies IgG1 b12(b12) could recognize conformational epitopes that overlap the CD4 bs of Env. Different virus strains, even derived from the same individual, showed distinct neutralization susceptibility to b12. We examined the key amino acid residues affecting b12 neutralization susceptibility using single genome amplification and pseudovirus neutralization assay. Eleven amino acid residues were identified that affect the sensitivity of Env to b12. Through site-directed mutagenesis, an amino acid substitution at position 182 in the V2 region of Env was confirmed to play a key role in regulating the b12 neutralization susceptibility. The introduction of V182 L to a resistant strain enhanced its sensitivity to b12 more than twofold. Correspondingly, the introduction of L182 V to a sensitive strain reduced its sensitivity to b12 more than tenfold. Amino acid substitution at positions 267 and 346 could both enhance the sensitivity to b12 more than twofold. However, no additive effect was observed when the three site mutageneses were introduced into the same strain, and the sensitivity was equivalent to the single V182 L mutation. CRF07_BC is a major circulating recombinant form of hiv-1 prevalent in China. Our data may provide important information for understanding the molecular mechanism regulating the neutralization susceptibility of CRF07_BC viruses to b12 and may be helpful for a vaccine design targeting the CD4 bs epitopes.

关键词： human immunodeficiency virus type 1 CRF07_BC envelope glycoprotein IgG1b12 neutralizing antibody single genome amplification

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Synthesis and biological evaluation of novel tricyclic matrinic derivatives as potential anti-filovirus agents

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Acta Pharmaceutica Sinica B 2018年第4期8卷 629-638页

作者： Xin Zhang Qiang Liu Qianqian Li Yinghong Li Zhandong Liu Hongbin Deng Sheng Tang Yanxiang Wang Youchun Wang Danqing Song Beijing Key Laboratory of Antimicrobial Agents Institute of Medicinal BiotechnologyChinese Academy of Medical Science & Peking Union Medical College division of hiv/aids and Sex-transmitted virus vaccines National Institutes for Food and Drug Control Department of Neurology Beijing Friendship HospitalCapital Medical University

Twenty-six novel tricyclic sophoridinic and matrinic derivatives containing a common chlorinated benzene fragment were designed, synthesized and evaluated for their anti-ebolavirus(EBOV)activities. Structure–activity relationship analysis indicated:(i) 12 N-dichlorobenzyl motif was beneficial for the activity;(ii) the chiral configuration at C5 atom might not affect the activity much. Among the target compounds, compound 7d exhibited the most potent potency against EBOV with an IC_(50) value of 5.29 μmol/L and an SI value of over 37.8. Further in vivo anti-EBOV assay of 7d identified its high effectiveness, and in vivo anti-MARV assay of 7d suggested its inspiring broad-spectrum anti-filovirus activity. The results provided powerful information on further strategic optimization and development of this kind of compounds against filoviruses.

关键词： Sophoridinic Matrinic Structure-activity relationship EBOV Filovirus

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Monkeypox virus quadrivalent mRNA vaccine induces immune response and protects against vaccinia virus

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Signal Transduction and Targeted Therapy 2023年第5期8卷 2423-2433页

作者： Ye Sang Zhen Zhang Fan Liu Haitao Lu Changxiao Yu Huisheng Sun Jinrong Long Yiming Cao Jierui Mai Yiqi Miao Xin Wang Jiaxin Fang Youchun Wang Weijin Huang Jing Yang Shengqi Wang Bioinformatics center of AMMS Beijing100850P.R.China division of hiv/aids and Sex-transmitted virus vaccines Institute for Biological Product ControlNational Institutes for Food and Drug Control(NIFDC)Beijing102629P.R.China Institute of Medical Biology Chinese Academy of Medical SciencesKunming650031P.R.China

Monkeypox has been declared a public health emergency by the World Health *** is an urgent need for efficient and safe vaccines against the monkeypox virus(MPXV)in response to the rapidly spreading monkeypox *** the age of COVID-19,mRNA vaccines have been highly successful and emerged as platforms enabling rapid development and large-scale ***,we develop two MPXV quadrivalent mRNA vaccines,named mRNA-A-LNP and mRNA-B-LNP,based on two intracellular mature virus specific proteins(A29L and M1R)and two extracellular enveloped virus specific proteins(A35R and B6R).By administering mRNA-A-LNP and mRNA-B-LNP intramuscularly twice,mice induce MPXV specific IgG antibodies and potent vaccinia virus(VACV)specific neutralizing ***,it elicits efficient MPXV specific Th-1 biased cellular immunity,as well as durable effector memory T and germinal center B cell responses in *** addition,two doses of mRNA-A-LNP and mRNA-B-LNP are protective against the VACV challenge in ***,the passive transfer of sera from mRNA-A-LNP and mRNA-B-LNP-immunized mice protects nude mice against the VACV ***,our results demonstrate that mRNA-A-LNP and mRNA-B-LNP appear to be safe and effective vaccine candidates against monkeypox epidemics,as well as against outbreaks caused by other orthopoxviruses,including the smallpox virus.

关键词： doses protective immunity

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Screening and identification of HTNV_(pv)entry inhibitors with high-throughput pseudovirus-based chemiluminescence

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Virologica Sinica 2022年第4期37卷 531-537页

作者： Xiaojing Wen Li Zhang Qiang Liu Xinyue Xiao Weijin Huang Youchun Wang division of hiv/aids and Sex-transmitted virus vaccines Institute for Biological Product ControlNational Institutes for Food and Drug Control(NIFDC)and WHO Collaborating Center for Standardization and Evaluation of BiologicalsBeijing 102629China Department of Microbiological Laboratory Technology School of Public HealthShandong UniversityJinan 250012China Institute for Reference Standards and Standardization National Institutes for Food and Drug ControlBeijing 102629China

Hantaviruses,such as Hantaan virus(HTNV)and Seoul virus,are the causative agents of Hantavirus cardiopulmonary syndrome(HCPS)and hemorrhagic fever with renal syndrome(HFRS),and are important zoonotic *** has the highest incidence of HFRS,which is mainly caused by HTNV and Seoul *** approved antiviral drugs are available for these hantaviral ***,a chemiluminescence-based highthroughput-screening(HTS)assay was developed and used to screen HTNV pseudovirus(HTNVpv)inhibitors in a library of 1813 approved drugs and 556 small-molecule compounds from traditional Chinese medicine *** identified six compounds with in vitro anti-HTNVpvactivities in the low-micromolar range(EC50values of0.1–2.2μmol/L;selectivity index of 40–900).Among the six selected compounds,cepharanthine not only showed good anti-HTNVpvactivity in vitro but also inhibited HTNVpv-fluc infection in Balb/c mice 5 h after infection by94%(180 mg/kg/d,Pviruses.

关键词： Hantaan virus(HTNV) High-throughput screening(HTS) Inhibitor Drug repurposing

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Cathepsin L plays a key role in SARS-CoV-2 infection in humans and humanized mice and is a promising target for new drug development

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Signal Transduction and Targeted Therapy 2021年第4期6卷 1357-1368页

作者： Miao-Miao Zhao Wei-Li Yang Fang-Yuan Yang Li Zhang Wei-Jin Huang Wei Hou Chang-Fa Fan Rong-Hua Jin Ying-Mei Feng You-Chun Wang Jin-Kui Yang Department of Endocrinology Beijing Tongren HospitalCapital Medical UniversityBeijingChina division of hiv/aids and Sex-transmitted virus vaccines Institute for Biological Product ControlNational Institutes for Food and Drug Control(NIFDC)BeijingChina Department of Science and Technology Beijing Youan HospitalCapital Medical UniversityBeijingChina division of Animal Model Research Institute for Laboratory Animal ResourcesNational Institutes for Food and Drug ControlBeijingChina

To discover new drugs to combat COVID-19,an understanding of the molecular basis of SARS-CoV-2 infection is urgently ***,for the first time,we report the crucial role of cathepsin L(CTSL)in patients with *** circulating level of CTSL was elevated after SARS-CoV-2 infection and was positively correlated with disease course and ***,SARS-CoV-2 pseudovirus infection increased CTSL expression in human cells in vitro and human ACE2 transgenic mice in vivo,while CTSL overexpression,in turn,enhanced pseudovirus infection in human *** functionally cleaved the SARS-CoV-2 spike protein and enhanced virus entry,as evidenced by CTSL overexpression and knockdown in vitro and application of CTSL inhibitor drugs in ***,amantadine,a licensed anti-influenza drug,significantly inhibited CTSL activity after SARS-CoV-2 pseudovirus infection and prevented infection both in vitro and in ***,CTSL is a promising target for new anti-COVID-19 drug development.

关键词： infection drugs ACE2

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Cellular tropism and antigenicity of mink-derived SARS-CoV-2 variants

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Signal Transduction and Targeted Therapy 2021年第6期6卷 1668-1670页

作者： Li Zhang Qianqian Li Jianhui Nie Ruxia Ding Haixin Wang Jiajing Wu Xuguang Li Xiaoming Yang Weijin Huang Youchun Wang division of hiv/aids and Sex-transmitted virus vaccines Institute for Biological Product ControlNational Institutes for Food and Drug Control(NIFDC)and WHO Collaborating Center for Standardization and Evaluation of BiologicalsBeijingChina Centre for Biologies Evaluation Biologic and Radiopharmaceutical Drugs DirectorateHPFBHealth CanadaOttawaONCanada China National Biotec Group Company Limited BeijingChina

Dear Editor,The outbreak of SARS-CoV-2 in minks has been observed recently,raising serious concerns over cross-species transmission and the emergence of variants capable of rendering antibody therapy and vaccines less ***,the species tropism and antigenicity of the spike protein of ten variants were analyzed in pseudovirus-based assays involving 25 cell lines as well as 293T cells expressing ACE2 receptor from 14 species.

关键词： ACE2 mink variants

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Animal models for COVID-19:advances,gaps and perspectives

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Signal Transduction and Targeted Therapy 2022年第8期7卷 2817-2840页

作者： Changfa Fan Yong Wu Xiong Rui Yuansong Yang Chen Ling Susu Liu Shunan Liu Youchun Wang division of Animal Model Research Institute for Laboratory Animal ResourcesNational Institutes for Food and Drug Control(NIFDC)National Rodent Laboratory Animal Resources CenterBeijing 102629China Department of Microbiology&Infectious Disease Center School of Basic Medical SciencesPeking University Health Science CenterBeijing 100083China College of Life Sciences Northwest UniversityProvincial Key Laboratory of Biotechnology of Shaanxi ProvinceNorthwest UniversityXi’an 710069China division of hiv/aids and sexually transmitted virus vaccines Institute for Biological Product ControlNational Institutes for Food and Drug Control(NIFDC)BeijingChina

COVID-19,caused by SARS-CoV-2,is the most consequential pandemic of this *** the outbreak in late 2019,animal models have been playing crucial roles in aiding the rapid development of vaccines/drugs for prevention and therapy,as well as understanding the pathogenesis of SARS-CoV-2 infection and immune responses of ***,the current animal models have some deficits and there is an urgent need for novel models to evaluate the virulence of variants of concerns(VOC),antibodydependent enhancement(ADE),and various comorbidities of *** review summarizes the clinical features of COVID-19 in different populations,and the characteristics of the major animal models of SARS-CoV-2,including those naturally susceptible animals,such as non-human primates,Syrian hamster,ferret,minks,poultry,livestock,and mouse models sensitized by genetically modified,AAV/adenoviral transduced,mouse-adapted strain of SARS-CoV-2,and by engraftment of human tissues or *** understanding the host receptors and proteases is essential for designing advanced genetically modified animal models,successful studies on receptors and proteases are also *** improved alternatives for future mouse models are proposed,including the reselection of alternative receptor genes or multiple gene combinations,the use of transgenic or knock-in method,and different strains for establishing the next generation of genetically modified mice.

关键词： prevention drugs establishing

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