Engineering of broadly reactive seroantibodies

广泛反应性血清抗体的工程

• 批准号: 10436784
• 负责人: Mohammad Mohseni Sajadi
• 金额: --
• 依托单位: BALTIMORE VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10436784
• 关键词: AIDS prevention; Antibodies; Antibody Therapy; Antigens; Autoimmune; Autoimmune Diseases; Autologous; Award; Caring; Categories; Cells; Chronic Disease; Clinical; Communicable Diseases; Data; Drug Kinetics; Engineering; Family; Genetic Variation; Goals; HIV; HIV Antibodies; HIV Envelope Protein gp120; HIV Infections; HIV-1; HIV-1 vaccine; Half-Life; Health; Healthcare; Human; IgG1; In Vitro; Infection; Infection prevention; Inflammatory; Knowledge; Lead; Life; Macaca mulatta; Malignant Neoplasms; Measures; Mediating; Medicine; Methods; Modality; Modeling; Monoclonal Antibodies; Mucous Membrane; Mutation; Passive Immunization; Persons; Plasma; Prevention; Preventive vaccine; Prophylactic treatment; Provider; Resistance; Savings; Sequencing Biochemistry; Series; Source; Specificity; Technology; Testing; Vaccines; Variant; Veterans; Viral; Virus; Work; antibody engineering; antibody mimetics; antiretroviral therapy; base; chronic infection; curative treatments; design; effective therapy; falls; humanized mouse; improved; in silico; in vivo; in vivo Model; infancy; lead candidate; member; monoclonal antibody production; mouse model; mutant; neonatal Fc receptor; neutralizing antibody; novel; prevent; resistance mutation; screening panel; success; vaccinology

Background/Rationale: Currently, there is no effective preventative vaccine for human immunodeficiency virus (HIV)-1, nor is there a cure for this chronic infection. At least one in 250 Veterans is infected with HIV. HIV-1 Clade B strains are the predominant strain in the US, and the overwhelming source of infection for Veterans (97% Clade B). The only treatment available is life-long therapy with combined anti-retroviral therapy. As part of our current Merit Award, we were successful in discovering a new family of broadly neutralizing antibodies (bNAbs) that are the most potent and broad described to date (N49 P series), including N49P7 (100% breadth in 117 virus panel, median IC50 0.10 ug/ml) and N4P9.3 (97% breadth in the 117 virus panel, median IC50 0.0495 ug/ml). Further, we identified HIV-1 envelope sequences from contemporaneous, circulating Clade B viruses from the same donor, thus delineating their resistance mutations. Other preliminary data include in silico data demonstrating antibody engineering, and in vivo data demonstrating the mouse models used. Objectives: Our hypothesis is that we can exploit both antibody and viral envelope sequences to engineer the N49 P series bNAbs towards even greater breadth and potency, thus covering variants with the potential to escape naturally occurring bNAbs of N49 and similar specificities. The specific aims of this proposal are 1) Engineer the N49 P series bNAbs to optimize clinical utility; and 2) Test the engineered bNAbs in two humanized mice models to establish in vivo efficacy against HIV-1. Methods: Our plan is to first develop a panel of engineered 60 mAbs from the N49 P series bNAbs. Engineering will be informed by crystallographic information regarding wild type N49P7 and N49P9.3 Fab- gp120 interactions; in silico modeling of N49P7 and N49P9.3 resistant Env variants found in standard screening panels; in silico modeling of N49P7 and N49P9.3 resistant Env variants found in the contemporaneous plasma that harbored the bNAb lineages. The aim will be to determine the key residues that are involved in neutralization (with a focus on Clade B strains), as well as those that are detrimental to stability. With this knowledge, we will create 60 mutants with the aim of improving potency/breadth, while maintaining or improving stability, lack of auto/polyreactivity, and ADCC. A list of 5 final bNAbs (in LS format) will be tested in humanized mice models for half-life, and in vivo efficacy (autologous and heterologous challenges in cell-free and cell-associated challenges in the Hu-PBL mouse model). Findings: Anticipated results will be isolation of new engineered bNAbs from the N49 P series family that will demonstrate improved potency and breadth, and adequate stability and PK. Furthermore, these bNAbs will be tested in a humanized mouse models for in vivo efficacy, further moving these products toward clinical use. Status: We have isolated the N49 P series of bNAbs. The team we have assembled has expertise in monoclonal antibody production, HIV-1 virus sequencing, biochemistry and antibody engineering, and mouse models of HIV-1 infection. Impact: If our hypothesis proves correct, these bNAbs will represent a clinical breakthrough and the lead candidate for antibodies being considered for HIV-1 prophylaxis and treatment, and could be used in a variety of clinical formats, especially at the VA, as HIV continues to be a problem for our Veterans.

背景/原理：目前，没有有效的预防性疫苗用于人类免疫缺陷病毒 （HIV）-1，也没有治愈这种慢性感染的方法。每250名退伍军人中至少有一人感染艾滋病毒。HIV-1 进化枝B菌株是美国的主要菌株，也是退伍军人感染的主要来源 (97%进化枝B）。唯一可用的治疗方法是终身治疗和抗逆转录病毒综合疗法。一部分 在我们目前的优异奖中，我们成功地发现了一个新的广泛中和抗体家族， 迄今为止描述的最有效和最广泛的bNAb（N49 P系列），包括N49 P7（100%宽度 117病毒组中，中位IC 50为0.10 ug/ml）和N4P9.3（117病毒组中，97%宽度，中位IC 50为 0.0495微克/毫升）。此外，我们从同期的循环进化枝B中鉴定了HIV-1包膜序列 来自同一供体的病毒，从而描述它们的抗性突变。其他初步数据包括 证明抗体工程化的计算机数据和证明所用小鼠模型的体内数据。 目的：我们的假设是，我们可以利用抗体和病毒包膜序列来工程化 N49 P系列bNAb具有更大的广度和效力，从而涵盖了具有潜在的 逃避N49和类似特异性的天然存在的bNAb。本建议的具体目标是：1） 工程化N49 P系列bNAb以优化临床效用;和2）在两个实验中测试工程化的bNAb 人源化小鼠模型以建立针对HIV-1的体内功效。 方法：我们的计划是首先从N49 P系列bNAb开发一组工程化的60种mAb。 将通过关于野生型N49 P7和N49P9.3 Fab的晶体学信息通知工程部。 gp 120相互作用;在标准品中发现的N49 P7和N49P9.3耐药Env变体的计算机模拟 筛选组;在N49 P7和N49P9.3中发现的耐药Env变体的计算机模拟 同时的血浆，窝藏bNAb谱系。目的是确定关键的残留物， 参与中和（重点是进化枝B菌株），以及对稳定性有害的那些。 有了这些知识，我们将创造60个突变体，目的是提高效力/广度，同时保持或 改善稳定性、缺乏自身/多反应性和ADCC。将对5个最终bNAb（LS格式）的列表进行检测， 用于半衰期和体内功效的人源化小鼠模型（在无细胞环境中的自体和异源激发） 和Hu-PBL小鼠模型中的细胞相关攻击）。 结果：预期结果将是从N49 P系列家族中分离出新的工程化bNAb， 显示出改善的效力和广度，以及足够的稳定性和PK。此外，这些bnab将 在人源化小鼠模型中测试体内功效，进一步将这些产品推向临床应用。 状态：我们已分离出N49 P系列bNAb。我们组建的团队在以下方面拥有专业知识： 单克隆抗体生产，HIV-1病毒测序，生物化学和抗体工程，以及小鼠 HIV-1感染模型。 影响：如果我们的假设被证明是正确的，这些bNAb将代表一个临床突破， 被认为是HIV-1预防和治疗抗体的候选者，并可用于各种 临床形式，特别是在VA，因为艾滋病毒仍然是我们退伍军人的一个问题。