Rational Design of Live Attenuated Influenza A Vaccine Candidates

描述（由申请人提供）：尽管有流感疫苗，但仅在美国每年就有大约35,000人死于流感和流感相关并发症。流感的临床影响促使人们寻找能够快速设计和容易生产的更有效的新型疫苗。我们已经开发了一种合理的、计算机辅助的方法来构建有效的流感病毒减毒活疫苗候选物。这一过程被称为合成减毒病毒工程（SAVE） （Coleman等人，2008年）。Science 320(5884):1784-7) SAVE技术中使用的定制设计衰减通过密码子对偏差的基因组尺度变化来操作（Mueller等人，2010年）。自然生物技术28:723-727)。在本提案中，我们将把我们在实验室菌株a/ PR8/3/34 （Mueller et al. 2010）中成功研制出减毒活疫苗的技术扩展到临床意义重大的大流行性流感菌株a/ California/07/2009 （CA07）。CA07是导致2009年H1N1（“猪流感”）大流行的毒株。将在小鼠模型中测试这些save减毒（减弱）CA07病毒的安全性水平及其作为活疫苗候选物的潜力。我们系统的衰减是通过在病毒基因组中设计数百个同义核苷酸变化来实现的。这种合理的基因设计方法使Codagenix公司能够保持氨基酸序列对所有病毒蛋白具有100%的同一性。这数百种突变通过高遗传稳定性为候选疫苗提供了广泛的安全边际。此外，保持与疫苗靶点100%匹配的氨基酸序列，提供了save减毒疫苗的一个特点，这是目前任何可用的疫苗技术都不具备的。通过将SAVE扩展到具有临床重要性的菌株，以及构建各种合成设计，我们将展示

DESCRIPTION (provided by applicant): Despite the availability of influenza vaccines, approximately 35,000 individuals die each year in the US alone from influenza, and influenza-related complications. The clinical impact of Influenza motivates the search for new, more effective vaccines that can be rapidly designed and easily produced. We have developed a rational, computer-aided approach to construct efficacious live attenuated (weakened) influenza virus vaccine candidates. The process is termed Synthetic Attenuated Virus Engineering (SAVE), (Coleman et al., 2008. Science 320(5884):1784-7) The custom designed attenuation used in the SAVE technology operates through genome-scale changes in codon pair bias (Mueller et al., 2010. Nature Biotechnology 28:723-727). In this proposal, we will extend our technology, which successfully created a live-attenuated vaccine in the laboratory strain A/PR8/3/34 (Mueller et al. 2010), to a clinically significant pandemic influenza strain, A/California/07/2009 (CA07). CA07 was the strain responsible for the 2009 H1N1 ("swine flu") pandemic. These SAVE-attenuated (weakened) CA07 viruses will be tested for their level of safety and their potential as live vaccine candidates in a mouse model. Attenuation in our system is achieved via designing hundreds of synonymous nucleotide changes across the viral genome. This rational gene design approach allows Codagenix Inc. to maintain the amino acid sequences to have 100% identity to all viral proteins. These hundreds of mutations offers a wide margin of safety for vaccine candidates via high genetic stability. Also maintaining a 100% amino acid sequence match to the vaccine target, provides a characteristic of SAVE-attenuated vaccines that is not present in any currently available vaccine technology. By extending SAVE to a strain of clinical importance, as well as constructing various synthetic designs, we will show that our unique computer algorithms allow us construct viable vaccine candidates with the additional ability to tune the amount of attenuation predictably, similar to the volume on a car radio. Given that SAVE is a computer-aided approach the method can be applied rapidly to any emerging influenza virus in its entirety. The deliverable of this project is an improved live attenuated vaccine strain of the A/California/07/2009 lineage that has 100% amino acid identity to the target strain. This vaccine candidate will be taken into further clinical development in Phase II. PUBLIC HEALTH RELEVANCE: Vaccination has been humankind's main most robust defense against viral disease. We describe an entirely novel and rapid method to generate anti-Influenza A vaccine candidates that might prove applicable to most if not all Influenza A strains.