Development of Zika viral pseudoinfectious virus as Zika vaccine candidate

开发寨卡病毒假感染病毒作为寨卡候选疫苗

基本信息

批准号：
9536650
负责人：
XIAOWU PANG
金额：
$ 30万
依托单位：
TENGEN BIOMEDICAL CO.
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2021-04-22
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9536650
关键词：
Aedes Americas Animals Antigens Asia Attenuated Birth Brazil Capsid Proteins Cell Line Cells Complementary DNA Country Culicidae DNA Vaccines Dengue Dengue Virus Development Disabled Persons Disease Outbreaks Elements Endoplasmic Reticulum Epidemic FDA approved Flavivirus Formalin Foundations French Polynesia Genes Genome Goals Guillain-Barré Syndrome Harvest Helper Viruses Immunity In Vitro Industrialization Infant Infection Infectious Agent International Investigation Japanese Encephalitis Lead Length Lethal Dose 50 Location Medical Membrane Proteins Microcephaly Micronesia Mus Neuropathy Pan American Health Organization Peptide Hydrolases Peptide Signal Sequences Pharmaceutical Preparations Phase Preventive Process Production Protein C Proteins Public Health Recombinants Replicon Reporting Research Safety Series Serotyping Small Business Technology Transfer Research Structural Protein Subunit Vaccines Syndrome System Teratogens Titrations Uganda United States Vaccines Vero Cells Viral Viral Antigens Viral Genome Viral Proteins Viral Structural Proteins Viral Vaccines Virion Virus Virus Diseases West Nile virus Yellow Fever Yellow fever virus ZIKV infection Zika Virus Zika virus vaccine base chemical synthesis co-infection cost effective experience genetic manipulation immunogenicity in vivo inducible gene expression mouse model novel particle pathogen pre-clinical programs public health emergency public health priorities response vaccine candidate vector mosquito

项目摘要

Development of Zika viral pseudoinfectious virus as zika vaccine candidate Abstract Zika virus(ZIKV) is a mosquito-borne flavivirus that was first identified in Uganda in 1947. It then spread to Asia and Pacific. Recently, the explosive outbreaks in Americas and its implication in Guillan Barré (GBS) syndrome and the birth of microcephalic infants following maternal infection lead to WHO declared that ZIKV infection is “a Public Health Emergency of International Concern“. Currently, there is no specific treatment. Thus, developing a safe and effective vaccine for ZIKV infection is a high priority for public health. Here, we propose to develop a novel ZIKV vaccine candidate based on ZIKV pseudoinfectious virus (PIV) produced by incorporation of ZIKV subgenomic replicon into viral particle in stable packaging cells. The approach is based on our extensive experience of dengue virus (DENV) PIV investigation. In DENV PIV production system, the subgenomic replicon was constructed by deleting the capsid protein (C) gene from the DENV genome and optimizing the signal peptide sequence of pre- membrane protein (prM) to facilitate the formation of viral particles. Packaging cells were developed for inducible expression of a bi-protein Cpr, where the protein pr is the “pr” segment of viral protein prM that holds the protein C on the endoplasmic reticulum (ER). When the replicon was introduced into the packaging cells, protein C was released from the bi-protein Cpr by a replicon-encoded viral protease. Coordinate expression of viral structural proteins by the replicon and packaging cells led to the incorporation of the replicon into viral particle to produce DENV PIVs. Toward to develop a ZIKV PIV, we have constructed a full-length cDNA clone of ZIKV by chemical synthesis. Based upon these highly promising results, the Specific Aims of this Phase I STTR proposal are: 1) construction of subgenome for ZIKV 2) development of stable packaging cell lines based on FDA approved Vero cells, 3) rapid and efficient harvesting of ZIKV PIV from infected packaging lines, and 4) preliminary analysis of the safety and immunogenicity of the proposed vaccine in mice. Successful completion of this program will enable a Phase II research including development of a cost-effective GMP process for PIVs production at industrial scale and a pre-clinical animal study of immunity to ZIKV infection.

开发寨卡病毒假感染病毒作为寨卡候选疫苗抽象的寨卡病毒 (ZIKV) 是一种蚊媒黄病毒，于 1947 年在乌干达首次发现。随后传播最近，美洲爆发的疫情及其对吉兰·巴雷的影响（GBS）综合征和产妇感染后小头婴儿的出生导致世界卫生组织宣布寨卡病毒感染是“国际关注的突发公共卫生事件”。因此，开发安全有效的 ZIKV 感染疫苗是当务之急。在此，我们建议基于 ZIKV 开发一种新型 ZIKV 候选疫苗。将 ZIKV 亚基因组复制子掺入病毒颗粒中产生的假感染病毒 (PIV) 该方法基于我们在登革热病毒 (DENV) PIV 方面的丰富经验。在DENV PIV生产系统中，通过删除亚基因组复制子来构建。来自 DENV 基因组的衣壳蛋白 (C) 基因并优化前体的信号肽序列开发了促进病毒颗粒形成的膜蛋白（prM）。双蛋白 Cpr 的诱导表达，其中蛋白 pr 是病毒蛋白 prM 的“pr”片段当复制子被引入内质网 (ER) 时，它将蛋白质 C 保持在内质网 (ER) 上。在包装细胞中，蛋白 C 通过复制子编码的病毒蛋白酶从双蛋白 Cpr 中释放出来。复制子和包装细胞协调表达病毒结构蛋白导致将复制子整合到病毒颗粒中以产生 DENV PIV，为了开发 ZIKV PIV。在此基础上，我们通过化学合成构建了 ZIKV 的全长 cDNA 克隆。由于结果良好，第一阶段 STTR 提案的具体目标是：1）亚基因组的构建针对 ZIKV 2) 基于 FDA 批准的 Vero 细胞开发稳定的包装细胞系，3) 快速和从受感染的包装线中高效收获 ZIKV PIV，以及 4) 安全性初步分析以及所提出的疫苗在小鼠中的免疫原性将成功完成该计划。启用 II 期研究，包括为 PIV 生产开发具有成本效益的 GMP 工艺 ZIKV 感染免疫的工业规模和临床前动物研究。