A General Molecular Strategy for Attenuation of Human Pathogenic Arenaviruses

人类致病性沙粒病毒减毒的通用分子策略

• 批准号: 9217579
• 负责人: Juan C. de la Torre
• 金额: $ 24.06万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-05 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9217579
• 关键词: Adverse effects; Africa; Animal Model; Antigens; Area; Arenaviridae; Arenavirus; Attenuated; Attenuated Live Virus Vaccine; Cavia; Cell Line; Cells; Clinical; Clinical Trials; Competence; Consensus Sequence; Cultured Cells; Development; Dideoxy Chain Termination DNA Sequencing; Disease; Engineering; Exhibits; Future; Genetic; Genome; Goals; Growth; Heterophile Antigens; Human; Immune response; Immunity; Immunize; Individual; Infection; Interferons; Knowledge; Label; Lassa Fever; Lassa virus; Lujo virus; Lymphocytic choriomeningitis virus; Mediating; Modeling; Molecular; Morbidity - disease rate; Mus; Mutation; Old World Arenaviruses; Pathogenicity; Phenotype; Population; Production; Property; Protocols documentation; Public Health; Reassortant Viruses; Recombinants; Resources; Ribavirin; Serial Passage; South Africa; Symptoms; Testing; Travel; Vaccines; Viral; Viral Hemorrhagic Fevers; Virulence; Virus; Virus Diseases; attenuation; base; biodefense; clinically significant; combat; design; epidemiology study; genetic approach; in vivo; interest; metropolitan; mortality; mouse model; neglect; novel; novel strategies; pathogen; public health relevance; pyrosequencing; response; reverse genetics

 DESCRIPTION (provided by applicant): Several arenaviruses cause hemorrhagic fever (HF) disease in humans and pose a serious public health problem in their endemic regions. Thus, the Old World arenavirus (OWA) Lassa (LASV) infects several hundred thousand individuals yearly in West Africa resulting in a high number of Lassa fever (LF) cases associated with high morbidity and mortality. Moreover, evidence indicates that the worldwide-distributed OWA LCMV is a neglected clinically important human pathogen. In addition, several arenaviruses including LASV and LCMV pose a credible biodefense threat. Existing anti-arenaviral therapy is limited to an off-label use of ribavirin that is only partially effective, and there are no FDA-licensed, or currently in clinical trials, arenavirus vaccines. However, the MOPV/LASV reassortant (ML29) is a candidate live-attenuated vaccine (L-AttV) for LASV that has shown promising results in animal models. Nevertheless, as with many other traditional L-AttV, the mechanism of ML29 attenuation remains unknown, which raises concerns about the phenotypic stability of ML29 in response to additional mutations. The central goal of this application is to test the hypothesis that we can convert a pathogenic OWA into an attenuated form with features of L-AttV via replacement of its L IGR by a genetically defined synthetic S-like IGR (Ssyn). Our hypothesis is supported by our recent following findings: 1) We could rescued rLCMV(IGR/S-S) with the same S-IGR in both S and L genome segments, and found that it was highly attenuated in vivo. 2) Mice immunized with rLCMV(IGR/S-S) were protected against a lethal challenge with wild type (WT) LCMV. 3) We have obtained evidence that a high degree of sequence plasticity within the S IGR is compatible with virus viability, which supports the feasibility of engineering recombinant arenaviruses with a synthetic S IGR (Ssyn) in the L segment as a general molecular strategy for arenavirus attenuation. To test our hypothesis we propose to complete the following specific aims: Aim 1. Characterize rLCMV (IGR/S-Ssyn): We have rescued rLCMV(IGR/S-Ssyn) where a synthetic S IGR (Ssyn) substituted for the L-IGR. We will characterize rLCMV(IGR/S-Ssyn) in cultured cells and using the mouse model of LCMV infection test the hypothesis that rLCMV(IGR/S-Syn) displays key features for L-AttV. Aim 2: Generate and characterize rLASV(IGR/S-Ssyn): We will generate rLASV(IGR/S-Ssyn) and test the hypothesis that it is attenuated in vivo but able to induce protective immunity against a letha challenge with WT LASV in a well-established guinea pig model of LASV infection. Aim 3. Examine the stability of rLCMV(IGR/S-Ssyn): We will test the hypothesis that rLCMV(IGR/S-Ssyn) is genetically stable during multiplication under different growth conditions. The successful completion of this application will uncover a general molecular strategy for arenavirus attenuation that can facilitate a novel strategy to develop L-AttV to combat human pathogenic arenaviruses.



项目成果

A Lassa Virus Live-Attenuated Vaccine Candidate Based on Rearrangement of the Intergenic Region

• DOI: 10.1128/mbio.00186-20
• 发表时间: 2020-03-01
• 期刊: MBIO
• 影响因子: 6.4
• 作者: Cai, Yingyun;Iwasaki, Masaharu;de la Torre, Juan Carlos
• 通讯作者: de la Torre, Juan Carlos

Molecular Engineering of a Mammarenavirus with Unbreachable Attenuation.

具有不可破坏的减毒能力的乳腺病毒的分子工程。

• DOI: 10.1128/jvi.01385-22
• 发表时间: 2023
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Sakabe,Saori;Cubitt,Beatrice;Martinez-Sobrido,Luis;delaTorre,JuanC
• 通讯作者: delaTorre,JuanC