Creation of Stable Cleaved Trimers of the HIV-1 Envelope Protein

HIV-1 包膜蛋白的稳定切割三聚体的创建

• 批准号: 7120976
• 负责人: Robert G. Whalen
• 金额: $ 24.04万
• 依托单位: MAXYGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7120976
• 关键词: AIDS; antigens; genes; infection; library; neutralizing antibody; proteins

DESCRIPTION (provided by applicant): The HIV-1 epidemic has resulted in ~5 million new infections in 2004 for a total of 40 million people living with HIV/AIDS. Clinical trials have shown that HIV-1 infection cannot be prevented by immunization with subunits of naturally occurring viral envelope (Env) proteins. However, it is clear from experimental studies that neutralizing antibodies to the Env protein can protect primates from infection. Modified Env proteins that better expose the neutralizing epitopes are now required to accelerate vaccine development. Detailed structural studies of the HIV-1 and SIV Env glycoproteins have revealed that this molecule undergoes substantial changes when it is bound by CD4, which is the initial step of virus-cell interaction. A major objective of HIV vaccine research is to create stable structures that are based on the unliganded form of the trimeric Env molecule. Molecules that resemble a stabilized version of the mature envelope trimer on the virion surface and that better expose existing neutralizing epitopes are currently being investigated. However, the Env structure carried by circulating HIV-1 clearly does not induce broadly neutralizing antibodies in infected individuals. Therefore, whatever the benefits of using a trimeric Env complex as an immunogen for vaccination, the Env protein must still be modified to improve the immunogenicity of conserved neutralizing epitopes. We propose to use directed molecular evolution to accomplish 2 main objectives. In this Phase I SBIR proposal, we will first use in vitro multigene DNA recombination to create libraries of novel chimeric HIV env genes that will be screened to identify a number of stable, cleaved, trimeric forms of the HIV-1 Env ectodomain. Second, upon reaching the goals of the present work, we will evaluate the immunogenicity of these proteins in a subsequent Phase II SBIR proposal. This work will provide a rigorous assessment of the value of trimeric Env molecules as immunogens and will provide lead molecules for further development as vaccine candidates. We are ultimately targeting a commercial vaccine product that will be of value in the prevention of HIV/AIDS. The Specific Aims of this Phase I SBIR proposal are summarized below. They are designed to evaluate the feasibility of creating stable and soluble forms of the HIV-1 Env trimer. -Specific Aim #1: Create libraries of recombined env genes encoding the gp140 ectodomain; -Specific Aim #2: Identify and characterize at least 10 stable forms of fully cleaved Env trimers; -Specific Aim #3: Prepare monomeric gp120 from each of the stable, cleaved Env trimers.

描述（由申请人提供）：HIV-1流行病在2004年导致了约500万种新感染，共有4000万人患有艾滋病毒/艾滋病。临床试验表明，与天然发生的病毒包膜（ENV）蛋白的亚基免疫无法预防HIV-1感染。但是，从实验研究中可以明显看出，对ENV蛋白的中和抗体可以保护灵长类动物免受感染。现在需要更好地暴露中和表位的改良ENV蛋白来加速疫苗的发育。 HIV-1和SIV ENV糖蛋白的详细结构研究表明，该分子在受CD4绑定时会发生实质性变化，这是病毒细胞相互作用的第一步。 HIV疫苗研究的一个主要目标是创建基于三聚体分子的非配合形式的稳定结构。目前正在研究类似于稳定的包膜三聚体的稳定版本，并且目前正在研究更好地暴露现有中和表位的分子。但是，通过循环HIV-1携带的ENV结构显然不会引起受感染个体的大规模中和抗体。因此，无论将三聚合物Env复合物作为免疫原进行疫苗接种的好处，ENV蛋白仍然必须修改以改善保守中和表位的免疫原性。我们建议使用定向的分子进化来实现2个主要目标。在此阶段I SBIR提案中，我们将首先使用体外多基因DNA重组来创建新型嵌合HIV Env基因的库，这些库将被筛选以识别许多稳定，切割，三聚体的HIV-1 Encododomain。其次，在达到本工作的目标后，我们将在随后的II期SBIR提案中评估这些蛋白质的免疫原性。这项工作将对三聚体Env分子作为免疫原子的价值进行严格的评估，并将提供铅分子作为候选疫苗的进一步开发。我们最终将目标定位在预防艾滋病毒/艾滋病的商业疫苗产品中。该阶段I SBIR提案的具体目标如下总结。它们旨在评估创建HIV-1 Env Trimer的稳定和可溶形式的可行性。 - 特定的目标＃1：创建编码GP140 EctodoMain的重组ENV基因的库； - 特定目标＃2：识别和表征至少10种完全切割的Env三聚体的稳定形式； - 特定的目标＃3：从每个稳定的，切割的Env三聚体中准备单体GP120。

项目成果

Testing Densities and Refractive Indices of Extraterrestrial Ice Components Using Molecular Structures - Organic Compounds and Molar Refractions.

使用分子结构 - 有机化合物和摩尔折射测试地外冰成分的密度和折射率。

• DOI: 10.3847/1538-4357/ab6efa
• 发表时间: 2020
• 期刊: The Astrophysical journal
• 作者: Hudson,ReggieL;Loeffler,MarkJ;Ferrante,RobertF;Gerakines,PerryA;Coleman,FalviaM
• 通讯作者: Coleman,FalviaM

Solid-State Isomerization and Infrared Band Strengths of Two Conformational Isomers of Cyclopropanecarboxaldehyde, A Candidate Interstellar Molecule.

候选星际分子环丙烷甲醛的两种构象异构体的固态异构化和红外波段强度。

• DOI: 10.1021/acsearthspacechem.9b00058
• 发表时间: 2019
• 期刊: ACS earth & space chemistry
• 影响因子: 3.4
• 作者: Hudson,ReggieL;Coleman,FalviaM
• 通讯作者: Coleman,FalviaM

Identifying the wide diversity of extraterrestrial purine and pyrimidine nucleobases in carbonaceous meteorites.

• DOI: 10.1038/s41467-022-29612-x
• 发表时间: 2022-04-26
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Oba, Yasuhiro;Takano, Yoshinori;Furukawa, Yoshihiro;Koga, Toshiki;Glavin, Daniel P.;Dworkin, Jason P.;Naraoka, Hiroshi
• 通讯作者: Naraoka, Hiroshi