Improve the safety of an efficacious live-attenuated HIV-1 vaccine through unnatu

通过 unnatu 提高有效的 HIV-1 减毒活疫苗的安全性

基本信息

批准号：
8837571
负责人：
Jiantao Guo
金额：
$ 46.49万
依托单位：
UNIVERSITY OF NEBRASKA LINCOLN
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-05-01 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8837571
关键词：
Acquired Immunodeficiency Syndrome Address Amino Acids Amino Acyl-tRNA Synthetases Animal Model Antigens Attenuated Bacteria Biochemical Biomedical Research Bone Marrow Charge Clinic Clinical Clinical Trials Code Codon Nucleotides Communicable Diseases Development Generations Genome Goals HIV HIV Genome HIV-1 Health Human Immunization In Vitro Infection Kinetics Lead Life Liver Macaca mulatta Mammalian Cell Mediating Modality Modeling Mus Nature Nebraska Pan Genus Prevention Protein Biosynthesis Regulatory Element Research SIV Safety Technology Testing Thymus Gland Time Transfer RNA Triplet Multiple Birth Universities Vaccine Research Vaccines Viral Genome Virulence Virus clinical application design global health improved in vitro testing in vivo novel novel strategies pandemic disease pathogen prevent rectal spatiotemporal stem tool transmission process

项目摘要

DESCRIPTION (provided by applicant): A safe and effective human immunodeficiency virus type one (HIV-1) vaccine is urgently needed to stem worldwide AIDS pandemic, but still remains elusive. Among all the HIV-1 vaccine modalities developed and tested thus far, nef-deleted live-attenuated HIV-1 vaccines (¿nef-LAV) confer the best protection against HIV-1 acquisition in simian immunodeficiency virus/rhesus macaque model of HIV-1 infection; however, its safety concerns preclude its clinical use. We at University of Nebraska-Lincoln in the past years have developed an unnatural amino acid (UAA)-mediated blank codon suppression approach to control HIV-1 protein biosynthesis. Our approach entails the manipulation of essential protein biosynthesis through blank codon (codons that do not encode a natural proteinogenic amino acid, such as nonsense triplet codon and frameshift codon) suppression that is precisely controlled by UAA that is not found in nature. We applied this technology to HIV-1 vaccine research by introducing several sets of blank codons coding for UAA into virus genome to control HIV-1 replication. We successfully demonstrated that we could turn HIV-1 replication on and off in vitro. We hypothesize that our novel strategy to allow live-attenuated HIV-1 to replicate only in the presence of the three artificial regulatory elements that are not found in nature (novel ¿nef-LAV) will greatly improve the safety of conventional ¿nef-LAV while still maintains its good efficacy against HIV-1 acquisition. We therefore propose to test this hypothesis with three specific aims. Specific Aim 1, to control the replication of novel ¿nef-LAV in vitro, of which we will apply and further optimize blank codon suppression strategy and incorporate suppression tRNA-aaRS pair into the HIV-1 genome to control HIV-1 replication by supplementing or withdrawing UAA in mammalian cells in vitro. Specific Aim 2, to control novel ¿nef-LAV replication in vivo, of which we will compare and contrast the in vivo replication capacity and kinetics of the novel ¿nef-LAV that has passed multi-cycle replication tests in vitro in the Specific Aim 1 with conventional ¿nef-LAV using humanized- BLT mice. The results of the Specific Aim 2 will be used to further optimize the novel ¿nef-LAV virus design and development in the Specific Aim 1. Specific Aim 3, novel ¿nef-LAV in vivo protection, of which we will compare the protection rate of novel ¿nef-LAV vs. conventional ¿nef-LAV against HIV-1 rectal challenge using hu-BLT mice. Since our strategy can greatly improves the safety of ¿nef-LAV by providing additional safety locks on HIV-1, it makes the modified ¿nef-LAV practical in clinical trial. In addition, the approaches/tools that are developed and further optimized in this research can be applied to the generation of vaccines against other pathogenic viruses or bacteria, and therefore is expected to exert broader impact on vaccine research.

描述（由适用提供）：迫切需要一种安全有效的人类免疫缺陷病毒（HIV-1）疫苗来阻止全球促进全球大流行，但仍然难以捉摸。在迄今为止开发和测试的所有HIV-1疫苗模式中，NEF删除的实时侵占的HIV-1疫苗（„ NEF-LAV）会议是HIV-1感染的HIV-1的最佳保护，以防止HIV-1采集HIV-1的HIV-1感染模型；但是，其安全性涉及其临床使用。在过去的几年中，我们在内布拉斯加州林肯大学（Univers of Nebraska-Lincoln）开发了一种不自然的氨基酸（UAA）介导的空白密码子抑制方法来控制HIV-1蛋白质生物合成。我们的方法需要通过空白密码子（不编码天然蛋白氨基酸的密码子（例如无义的三重态密码子和Frameshift密码子）抑制）对必需蛋白质生物合成的操纵，这些密码由UAA精确控制，而UAA在本质上没有发现。我们将该技术应用于HIV-1疫苗研究中，通过将编码UAA编码的几组空白密码子引入病毒基因组中以控制HIV-1复制。我们成功证明了我们可以在体外打开和关闭HIV-1复制。我们假设我们的新型策略允许活体侵蚀的HIV-1仅在存在自然界中未发现的三种人工调节元素的情况下复制（新颖的„ NEF-LAV），这将大大提高常规„ NEF-LAV的安全性，而仍然保持其良好的效率，以抗HIV-1。因此，我们建议用三个特定目标检验这一假设。具体目的1，以控制新型``nef-LAV体外的复制，我们将应用并进一步优化空白的密码子抑制策略，并将抑制tRNA-AARS对纳入HIV-1基因组中，以通过在体外哺乳动物细胞中补充或撤回UAA来控制HIV-1的复制。具体目的2，用于控制新型的体内nef-LAV复制，我们将在其中比较和对比新颖的体内复制能力和动力学，而新颖的LAV的动力学已经通过特定的目标1中的多周期复制测试与常规的nef-lav使用人体化的nef-lav，使用人体化的nef-lav。特定目标2的结果将用于进一步优化特定目的的新型``nef-LAV病毒设计和开发。由于我们的策略可以通过在HIV-1上提供其他安全锁来大大提高`NEF-LAV的安全性，因此它使经过修改的`nef-LAV在临床试验中实用。此外，在本研究中开发并进一步优化的方法/工具可以应用于针对其他致病病毒或细菌的疫苗，因此有望对疫苗研究产生更大的影响。