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/理由：目前，人类免疫缺陷病毒尚无有效的预防疫苗 (HIV)-1，这种慢性感染也没有治愈方法。至少二分之一的退伍军人感染艾滋病毒。 HIV-1 B 分支菌株是美国的主要菌株，也是退伍军人的压倒性感染源 （97% 进化枝 B）。唯一可用的治疗方法是终身治疗与抗逆转录病毒联合治疗。作为一部分 在我们当前的优异奖中，我们成功地发现了一个新的广泛中和抗体家族 (bNAb)，是迄今为止描述的最有效和最广泛的（N49 P 系列），包括 N49P7（100% 宽度） 在 117 病毒组中，中值 IC50 0.10 ug/ml）和 N4P9.3（在 117 病毒组中，中值 IC50 为 97%） 0.0495微克/毫升）。此外，我们还从同时流行的 B 分支中鉴定出 HIV-1 包膜序列 来自同一供体的病毒，从而描绘出它们的抗性突变。其他初步数据包括 计算机数据证明了抗体工程，体内数据证明了所使用的小鼠模型。 目标：我们的假设是我们可以利用抗体和病毒包膜序列来设计 N49 P 系列 bNAb 具有更大的广度和效力，从而涵盖了具有潜力的变体 逃避天然存在的 N49 的 bNAb 和类似的特异性。该提案的具体目标是 1) 设计 N49 P 系列 bNAb 以优化临床效用； 2) 在两种情况下测试工程化 bNAb 人源化小鼠模型建立体内抗 HIV-1 功效。 方法：我们的计划是首先从 N49 P 系列 bNAb 中开发一组工程化的 60 种 mAb。 工程人员将了解有关野生型 N49P7 和 N49P9.3 Fab- 的晶体学信息 gp120 相互作用；标准中发现的 N49P7 和 N49P9.3 抗性 Env 变体的计算机建模 筛选板；在计算机模型中发现的 N49P7 和 N49P9.3 抗性 Env 变体 含有 bNAb 谱系的同期血浆。目的是确定关键残留物 参与中和（重点关注分支 B 菌株），以及那些对稳定性有害的菌株。 有了这些知识，我们将创建 60 个突变体，目的是提高效力/广度，同时保持或 提高稳定性、缺乏自体/多反应性和 ADCC。 5 个最终 bNAb 列表（LS 格式）将在 人源化小鼠模型的半衰期和体内功效（无细胞中的自体和异体挑战） 以及 Hu-PBL 小鼠模型中细胞相关的挑战）。 研究结果：预期结果是从 N49 P 系列家族中分离出新的工程 bNAb，这将 表现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了体内功效，进一步将这些产品推向临床应用。 状态：我们已经分离出 N49 P 系列 bNAb。我们组建的团队拥有以下专业知识 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 将代表临床突破并处于领先地位 候选抗体被考虑用于 HIV-1 预防和治疗，并可用于多种 临床形式，特别是在退伍军人管理局，因为艾滋病毒仍然是我们退伍军人的一个问题。