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

• 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
• 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

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（由申请人提供）：迫切需要一种安全有效的人类免疫缺陷病毒一型（HIV-1）疫苗来阻止世界范围内的艾滋病流行，但在迄今为止开发和测试的所有 HIV-1 疫苗模式中仍然难以捉摸。 nef 删除的 HIV-1 减毒活疫苗 (¿nef-LAV) 可在猿猴免疫缺陷病毒/恒河猴模型中提供针对 HIV-1 感染的最佳保护HIV-1 感染；然而，其安全性问题阻碍了其临床应用。过去几年，我们内布拉斯加大学林肯分校开发了一种非天然氨基酸 (UAA) 介导的空白密码子抑制方法来控制 HIV-1 蛋白质的生物合成。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，例如无义三联密码子和移码密码子抑制，由 UAA 精确控制，而不是）来操纵必需的蛋白质生物合成。我们将这一技术应用于HIV-1疫苗研究，通过将多组编码UAA的空白密码子引入病毒基因组来控制HIV-1复制，我们成功地证明了我们可以在体外打开和关闭HIV-1复制。我们追求的新策略是让减毒活病毒 HIV-1 仅在自然界中未发现的三种人工调控元件存在的情况下才能复制（新颖 ¿ nef-LAV）将大大提高常规¿ nef-LAV 同时仍保持其对抗 HIV-1 感染的良好功效，因此我们建议通过三个具体目标来检验这一假设：控制新型病毒的复制。 nef-LAV体外，其中我们将应用并进一步优化空白密码子抑制策略，将抑制tRNA-aaRS对整合到HIV-1基因组中，通过在体外补充或撤回UAA来控制HIV-1复制。 2、控制小说¿ nef-LAV在体内的复制，我们将比较和对比新型的体内复制能力和动力学 ¿ nef-LAV 已通过特定目标 1 中与常规 ¿ 的体外多周期复制测试使用人源化 BLT 小鼠的 nef-LAV 具体目标 2 的结果将用于进一步优化新型 ¿ nef-LAV病毒在Specific Aim 1中的设计和开发。Specific Aim 3，新颖¿ nef-LAV的体内保护作用，我们将比较其中新颖的保护率 ¿ nef-LAV 与传统 ¿ nef-LAV 使用 hu-BLT 小鼠对抗 HIV-1 直肠攻击，因为我们的策略可以大大提高 ¿ nef-LAV 通过对 HIV-1 提供额外的安全锁，使得修改后的 ¿此外，本研究开发和进一步优化的方法/工具可应用于针对其他病原病毒或细菌的疫苗的生产，因此有望对疫苗研究产生更广泛的影响。 。