Engineering of broadly reactive seroantibodies

广泛反应性血清抗体的工程

• 批准号: 9890151
• 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/9890151
• 关键词: AIDS prevention; Antibodies; Antibody Therapy; Antigens; Autoimmune Diseases; Autoimmune Process; 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; Proteins; Provider; Resistance; Savings; Sequencing Biochemistry; Series; Source; Specificity; Technology; Testing; Vaccines; Variant; Veterans; Viral; Virus; Work; antibody engineering; 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，人类免疫缺陷病毒没有有效的预防疫苗 （HIV）-1，这种慢性感染也没有治愈。至少有250名退伍军人感染了艾滋病毒。 HIV-1 进化枝B菌株是美国的主要菌株，也是退伍军人的压倒性感染来源 （97％进化枝B）。唯一可用的治疗方法是终身治疗抗逆转录病毒疗法。作为一部分 在我们当前的优点奖中，我们成功地发现了一个新的广泛中和抗体的家庭 （bnabs）迄今为止最有效，最广泛的描述（N49 p系列），包括N49P7（100％广度 在117个病毒面板中，中位数IC50 0.10 ug/ml）和N4P9.3（117个病毒面板中的宽度为97％，中位数IC50 0.0495 ug/ml）。此外，我们从同时循环进化枝B中确定了HIV-1信封序列 来自同一供体的病毒，从而描绘了其耐药性突变。其他初步数据包括 硅数据证明了抗体工程和体内数据，证明了所使用的小鼠模型。 目的：我们的假设是我们可以利用抗体和病毒信封序列来设计 N49 p系列bnabs朝着更大的宽度和效力，因此涵盖了具有潜力的变体 逃避N49的自然发生的BNAB和类似的特异性。该提案的具体目的是1） 工程师N49 P系列BNABS以优化临床实用程序； 2）测试两次工程的bnabs 人性化小鼠模型以建立对HIV-1的体内功效。 方法：我们的计划是首先从N49 P系列BNABS开发一个工程化的60个单克隆抗体。 有关野生型N49P7和N49P9.3 Fab- GP120相互作用；在N49P7和N49P9.3标准中发现的N49P7和N49P9.3抗性ENV变体中 筛选面板；在N49P7和N49P9.3抗性ENV变体中的硅硅建模中 同时的等离子体具有BNAB谱系。目的是确定关键残留物 参与中和（重点是进化枝B菌株），以及那些不利于稳定性的人。 有了这些知识，我们将创建60个突变体，以提高效力/宽度，同时保持或 提高稳定性，缺乏自动/多反应性和ADCC。将测试5个最终BNAB（以LS格式）的列表 人性化的小鼠模型，用于半衰期和体内功效（无细胞的自体和异源挑战 在HU-PBL小鼠模型中与细胞相关的挑战）。 调查结果：预期的结果将是将N49 P系列的新工程BNAB隔离 表现出提高的效力和广度，以及足够的稳定性和PK。此外，这些bnabs将是 在人源化的小鼠模型中测试了用于体内功效的模型，进一步将这些产品移至临床使用。 状态：我们已经隔离了N49 P系列BNAB。我们组装的团队具有专业知识 单克隆抗体产生，HIV-1病毒测序，生物化学和抗体工程以及小鼠 HIV-1感染的模型。 影响：如果我们的假设被证明是正确的，这些BNAB将代表临床突破和铅 考虑用于HIV-1预防和治疗的抗体的候选者，可用于多样性 临床格式，尤其是在VA，因为HIV仍然是我们退伍军人的问题。