Project 1: Establishing a robust functional cure

项目 1：建立强大的功能性治疗方法

• 批准号: 10381477
• 负责人: Mauricio de Aguiar Martins
• 金额: $ 50.87万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-15 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10381477
• 关键词: Address; Animals; Antibodies; Antibody Response; CD4 Antigens; Capsid; Cells; Collaborations; Data; Dependovirus; Disease remission; Dose; Epitopes; Evaluation; Excision; Fc domain; Glycoproteins; Goals; HIV Envelope Protein gp120; HIV-1; HIV-2; Human; Immune; In Vitro; Infection; Macaca; Macaca mulatta; Measures; Mediating; Monkeys; Mus; Patients; Pattern; Property; Prophylactic treatment; Proviruses; Reporting; Resistance; Resources; SIV; Safety; Serum; Testing; Therapeutic; Time; Vaccination; Variant; Viral; Viral reservoir; Virus; Virus Replication; antibody-dependent cell cytotoxicity; antiretroviral therapy; base; cost; design; fitness; immune clearance; immunogenic; immunogenicity; improved; mimetics; neutralizing antibody; peptidomimetics; promoter; simian human immunodeficiency virus; tyrosine O-sulfate; vector; vector genome

PROJECT SUMMARY (Project 1 – Robust functional cures with AAV-expressed eCD4-Ig) eCD4-Ig is a potent and exceptionally broad fusion of the first two domains of CD4 to an antibody Fc domain and a short tyrosine-sulfated coreceptor-mimetic peptide. In rhesus macaques, adeno-associated virus (AAV)-expressed eCD4-Ig mediates consistent and very effective protection against SHIV-AD8 and SIVmac239. eCD4-Ig also has properties that make it especially useful for establishing a functional cure in rhesus macaques and perhaps in humans. These include its potency, breadth, difficulty-of-escape, low immunogenicity when expressed by AAV, consistent expression by AAV, potent intrinsic ADCC activity, and collaboration with serum antibodies to mediate ADCC. These properties allow eCD4-Ig to circumvent two major problems associated with using AAV-expressed antibodies to establish functional cures, namely immune clearance and viral escape. In preliminary data we show that AAV-expressed eCD4-Ig can suppress replication of SHIV-AD8 for more than a year in 5 of 6 macaques. However we also show that, when compared to the “Monkey monkey” described in Project 2, this suppression was less robust, meaning that consistent ‘blipping’ of virus was observed in most eCD4-Ig-suppressed animals. Because the Miami monkey expresses roughly 10 times the amount of total antibody observed in these eCD4-Ig-expressing macaques, we hypothesize that greater expression of eCD4-Ig will result in more robust functional cures. The goals of this project are thus to increase AAV-mediated expression of eCD4-Ig, to establish robust functional cures in SHIV-AD8 and SIVmac239-infected macaques, to amplify the ADCC activities in these macaques by stimulating host antibody responses to epitopes unmasked by eCD4-Ig, to establish a consistent platform that will allow for evaluation of latency reversing agents, and to ask whether long-term expression of eCD4-Ig by itself can impact the viral reservoir. These studies will improve and help understand a viable approach to establishing similar functional cures in humans.

项目摘要（项目 1 – 使用 AAV 表达的 eCD4-Ig 进行稳健的功能性治疗） eCD4-Ig 是 CD4 前两个结构域与抗体 Fc 的有效且异常广泛的融合 域和短酪氨酸硫酸化辅助受体模拟肽在恒河猴中，腺相关。 病毒 (AAV) 表达的 eCD4-Ig 介导针对 SHIV-AD8 和 SIVmac239 还具有使其特别适用于建立功能性治愈的特性。 这些包括它的效力、广度、逃脱难度、低等。 AAV 表达时的免疫原性、AAV 的一致表达、有效的内在 ADCC 活性，以及 与血清抗体合作介导 ADCC，这些特性使 eCD4-Ig 能够规避两种情况。 与使用 AAV 表达的抗体建立功能性治疗相关的主要问题，即 初步数据表明，AAV 表达的 eCD4-Ig 可以促进免疫清除和病毒逃逸。 在 6 只猕猴中，有 5 只抑制 SHIV-AD8 的复制一年多。 与项目 2 中描述的“猴子”相比，这种抑制不那么强烈，这意味着 在大多数 eCD4-Ig 抑制动物中观察到病毒持续“闪烁”，因为迈阿密。 猴子表达的抗体量大约是这些表达 eCD4-Ig 的抗体总量的 10 倍 在猕猴中，我们追求 eCD4-Ig 的更高表达将带来更稳健的功能性治愈。 因此，该项目的目标是增加 AAV 介导的 eCD4-Ig 表达，建立稳健的 对 SHIV-AD8 和 SIVmac239 感染的猕猴进行功能性治愈，以增强这些猕猴中的 ADCC 活性 猕猴通过刺激宿主抗体对 eCD4-Ig 暴露的表位做出反应，建立 一致的平台，将允许评估延迟逆转代理，并询问是否长期 eCD4-Ig 的表达本身可以影响病毒库，这些研究将改善并提供帮助。 了解在人类中建立类似功能性治疗的可行方法。