Development of Zika viral pseudoinfectious virus as Zika vaccine candidate

谓词SBIR/STTR第一阶段拨款和团队执行摘要 Tengen Biomedical Co.和Howard University获得了STTR第一阶段资助(5R41AI129119-02 2017年，美国国家过敏和传染病研究所(NIAID)。我们的谓词的总体目标是 STTR第一阶段赠款是基于以下新概念开发一种安全、有效和负担得起的寨卡病毒疫苗 第三代黄病毒疫苗。寨卡病毒流行及其与寨卡病毒感染的关系 格林-巴利综合征和先天性残疾，包括小头畸形症，领导着世界卫生组织 2016年，世界卫生组织宣布ZIKV为“国际关注的突发公共卫生事件”。因此，一个 目前迫切需要有效的寨卡病毒疫苗。 该项目提出了四个具体目标： 1.ZIKV亚基因组复制子的构建 2.建立稳定的包装细胞系，提供反式ZIKV结构蛋白C。 3.从感染的包装细胞系中提取ZIKV PIVs，并对包装细胞的稳定性进行分析 在旅途中的线条和PIV， 4.初步分析该疫苗在小鼠体内的安全性和免疫原性。 两年来，我们在推进具体目标方面取得了8项重大成就： 1.寨卡病毒野生型和变异型感染性克隆的构建 2.选择适应建立稳定的脊椎动物特异性复制缺陷ZIKV 3.脊椎动物细胞中稳定复制缺陷基因ZIKV(VSRD-ZIKV)的DNA测序及分析 新的工程突变 4.VSRD-ZIKV在新生和3周龄免疫缺陷小鼠中的初步评价 5.Prime-Boost VSRD-ZIKV疫苗提供了强大的保护，以抵御致命的ZIKV挑战 免疫缺陷小鼠 6.VSRD-ZIKV疫苗接种与ZIKV特异性体液免疫和CD8+干扰素-+相关 回应 7.VSRD-ZIKV提供保护并防止病毒在挑战的雄性小鼠睾丸中积聚 用致命的ZIKV 8.VSRD-ZIKV免疫可保护孕鼠免受垂直传播 致命的ZIKV 此外，我们最近在此研究的基础上发表了一篇论文：万S、曹S、王旭、周勇、严伟、顾欣、吴 脊椎动物特异性复制缺陷寨卡病毒的产生及初步特征。 病毒学。2020年10月6日；552：73-82。DOI：10.1016/j.virol。2020.09.001。在线版先于印刷版。PMID：33075709 我们建议i-Corps团队包括三名高素质的关键调查人员。克里斯·D·塔，MBA。首席执行官 天根生物医药公司，拥有17年以上的行业开发经验；庞晓武， 博士，STTR第一阶段项目的首席研究员，他一直在推动该项目；以及顾新斌，医学博士。 博士，Tengen Biomedical Co.的联合创始人和STTR第一阶段项目的联合调查员。顾医生一直在 带领团队中的梦想家。所有三名成员都承诺遵守 程序。

Executive Summary of Predicate SBIR/STTR Phase I Grant and Team TenGen Biomedical Co. and Howard University received an STTR Phase I grant (5R41AI129119-02) from the National Institute of Allergy and Infectious Diseases (NIAID) in 2017. The overall objective of our predicate STTR Phase I grant is to develop a safe, effective, and affordable Zika vaccine based on the novel concept of the third generation of flavivirus vaccine. Zika virus (ZIKV) epidemics and the association of ZIKV infection with Guillain–Barré syndrome, and congenital disabilities, including microcephaly, led the World Health Organization to declare ZIKV a “Public Health Emergency of International Concern” in 2016. Therefore, an effective Zika vaccine is urgently needed. The project proposed four specific aims: 1. Construction of subgenomic replicon for ZIKV, 2. Development of stable packaging cell lines providing ZIKV structural protein C in trans. 3. Harvest of ZIKV PIVs from infected packaging cell line and analyzing the stability of the packaging cell lines and PIV during passages, 4. Preliminary analysis of the safety and immunogenicity of the proposed vaccine in mice. In the past two years, we have made 8 significant achievements forward the specific aims: 1. Generation of Infectious cDNA Clones of Wildtype and Mutant Zika Virus 2. Development of stable Vertebrate Specific Replication Defected ZIKV by selective adaptation 3. DNA sequencing of the stably replication-defective in vertebrate cells-ZIKV (VSRD-ZIKV) and analysis of new engineered mutations 4. Preliminary evaluation of VSRD-ZIKV in both newborn and 3-week-old immunocompromised mice 5. Prime-boost VSRD-ZIKV vaccination provides robust protection against lethal ZIKV challenge in immunocompromised mice 6. Vaccination with VSRD-ZIKV is associated with ZIKV-specific humoral immunity and CD8+IFN- + responses 7. VSRD-ZIKV provides protection and prevents viral accumulation in the testes of male mice challenged with lethal ZIKV 8. Immunization with VSRD-ZIKV protects against vertical transmission in pregnant mice challenged with lethal ZIKV In addition, we recently published a paper based on this study: Wan S, Cao S, Wang X, Zhou Y, Yan W, Gu X, Wu TC, Pang X. Generation and preliminary characterization of vertebrate-specific replication-defective Zika virus. Virology. 2020 Oct 6;552:73-82. doi: 10.1016/j.virol. 2020.09.001. Online ahead of print. PMID: 33075709 We proposed the I-Corps team included three highly qualified key investigators. Chris D. Ta, MBA. CEO of TenGen Biomedical Co. and he has over seventeen years of experience in industry development; Xiaowu Pang, Ph.D. Principal Investigator of the STTR Phase I project and he has been driving the project; and Xinbin Gu, MD. Ph.D. Co-founder of TenGen Biomedical Co. and Co-Investigator of the STTR Phase I project. Dr. Gu has been leading the visionary on the team. All three members are committed to the time requirements of the program.