Defining the impact of glycan-microdomains of the HIV-1 Env glycan shield

定义 HIV-1 Env 聚糖屏蔽的聚糖微结构域的影响

• 批准号: 10586068
• 负责人: JAN NOVAK
• 金额: $ 71.86万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-10 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10586068
• 关键词: Affect; Apical; Area; Binding Sites; Bioinformatics; Biological; Biology; Complex; Data; Databases; Engineering; Epitopes; Exhibits; Experimental Models; Future; Glycoproteins; Goals; HIV; HIV Envelope Protein gp120; HIV-1; Heterogeneity; Human Biology; Immune; Immune response; Immune system; Infection; Informatics; Intervention; Mannose; Methodology; Modeling; Molecular; Molecular Biology; Mutate; N Domain; N-Glycosylation Site; Penetration; Persons; Polysaccharides; Positioning Attribute; Process; Publications; Publishing; Recombinants; Research; Role; Sequence Analysis; Series; Site; Structure-Activity Relationship; Surface; Surface Antigens; Tertiary Protein Structure; Testing; Therapeutic; Time; Variant; Virus; Virus Replication; Work; design; dimer; experimental study; glycosylation; insight; molecular dynamics; molecular mass; neutralizing antibody; novel; pathogen; prevent; receptor; tool; viral transmission

ABSTRACT Human immunodeficiency virus 1 (HIV-1) is a major global pathogen, with 1.7 million new infections in 2019. The HIV-1 envelope glycoprotein (Env) is the sole virus-surface antigen. Env, a trimer of gp120/gp41 dimer subunits, is densely glycosylated by N-glycans. For the gp120 subunit, N-glycans contribute over 50% of its molecular mass and form a protective surface, the so-called “glycan shield”, that interacts with cellular receptors and the immune system and protects the virus from neutralizing antibodies (nAbs). Recently, the concept of an “evolving glycan shield” has been proposed, based on the sequence analysis of immune-escape variants that have lost some potential N-glycosylation sites (NGS) and gained others. Despite significant strides in understanding the biology of HIV-1 Env, the complexity and function of N-glycans at a molecular level are poorly understood. Based on our bioinformatics and experimental studies, we recently proposed that the Env glycan shield may be defined, and analyzed, as distinct structural glycan-microdomains. Using naturally occurring Env variants of a clade B virus, we analyzed the glycans surrounding the highly conserved N262 glycan, in the so- called high-mannose patch (HMP) near the apex of the Env trimer. These HMP glycans, comprised of different combinations of 4 to 6 NGS in the proximity of N262, appear to form a glycan working unit. Based on our published results and preliminary data, we hypothesize that Env outer domain, gp120 glycan shield, has structural components that can be defined as distinct structural glycan-microdomains. We propose to test our hypothesis in different clades of HIV-1 and their immune-escape variants by assessing structural and functional impacts of a range of N-glycan combinations that comprise Env glycan-microdomains. Specifically, we propose to determine how different NGS combinations of the N262-anchored HMP-microdomain across variants and clades influence Env functions (Aim 1), to probe the structural arrangement of different N262-glycan cluster NGS combinations in the context of functional recombinant Env trimers (Aim 2), and to assess the functional impacts of other potential glycan-clusters of the HIV-1 Env glycan shield (Aim 3). To accomplish these goals, we will use a combination of functional, bioanalytical, structural bioinformatics, and molecular dynamics simulation approaches. The results of these studies will test our proposed hypothesis on the functional impact of the structural components of the glycan shield, glycan-microdomains, and will determine how the varied compositions of these microdomains affect the function of the glycan shield. These data will advance our understanding of the molecular biology of HIV-1 and the factors that impact viral transmission and persistence. Ultimately, these studies will generate new information about Env vulnerabilities that can be exploited to develop future therapeutic approaches.

抽象的 人类免疫缺陷病毒1（HIV-1）是一种主要的全球病原体，2019年有170万个新感染。 HIV-1包膜糖蛋白（ENV）是唯一的病毒表面抗原。 Env，GP120/GP41二聚体的触发器 亚基被N-聚糖脱糖基化。对于GP120亚基，N-聚糖贡献了超过50％ 分子质量并形成一个受保护的表面，即所谓的“聚糖盾牌”，与细胞接收器相互作用 和免疫系统并保护病毒免受中和抗体（NABS）的侵害。最近， 基于免疫escape变体的序列分析，已经提出了“不断发展的聚糖屏蔽”。 已经失去了一些潜在的N-糖基化位点（NGS），并获得了其他潜在的。尽管大步很大 了解HIV-1 Env的生物学，N-聚糖在分子水平上的复杂性和功能很差 理解。根据我们的生物信息学和实验研究，我们最近提出了ENV Glycan 可以将盾牌定义并分析为不同的结构性聚糖 - 微域。使用自然发生的Env 进化枝B病毒的变体，我们分析了高度保守的N262 Glycan周围的聚糖， 在env触发器的顶点附近称为高甘露糖贴片（HMP）。这些HMP聚糖由不同的 N262的接近度中4至6 ng的组合似乎形成了一个聚糖工作单元。基于我们 已发布的结果和初步数据，我们假设ENV外域GP120 Glycan Shield具有 可以定义为不同结构性聚糖 - 微域的结构成分。我们建议 通过评估结构和 一系列N-Glycan组合的功能影响，包括Env Glycan-Microdomain。具体来说， 我们建议确定N262锚定的HMP-Microdomain的不同NGS组合 变体和进化枝会影响设想功能（目标1），以探测不同N262-Glycan的结构排列 在功能重组ENV三聚体的背景下（AIM 2）的群集NGS组合，并评估 HIV-1 Env Glycan盾牌的其他潜在聚糖群体的功能影响（AIM 3）。完成这些 目标，我们将使用功能，生物分析，结构生物信息学和分子动力学的组合 模拟方法。这些研究的结果将检验我们提出的关于功能影响的假设 聚糖屏蔽层的结构成分，聚糖 - 微域，并将确定如何变化 这些微区域的组成会影响聚糖屏蔽的功能。这些数据将推动我们的 了解HIV-1的分子生物学以及影响病毒传播和持久性的因素。 最终，这些研究将生成有关ENV漏洞的新信息，可以探索以开发 未来的治疗方法。