Rational Structure-Based Design of Broad Neutralizing Humanized svMP mAbs

基于合理结构的广泛中和人源化 svMP 单克隆抗体的设计

• 批准号: 10310508
• 负责人: Xin Ge
• 金额: $ 18.39万
• 依托单位: TEXAS A&M UNIVERSITY-KINGSVILLE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-02 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10310508
• 关键词: ADAMTS; Active Sites; Acute; Address; Affect; Affinity; Allergic Reaction; American; Animals; Antibodies; Antibody Binding Sites; Antidotes; Antivenins; Basement membrane; Binding; Biochemical; Biological Assay; Bite; Blood Circulation; Blood Coagulation Disorders; Blood Coagulation Factor; Blood capillaries; Blood coagulation; Bulla; California; Catalytic Domain; Cessation of life; Collaborations; Consensus; Consensus Sequence; Copperheads; Cross Reactions; Crotalus; Data; Development; Disabled Persons; Disseminated Intravascular Coagulation; Domestic Animals; Endothelial Cells; Engineering; Evaluation; Exploratory/Developmental Grant for Diagnostic Cancer Imaging; Extravasation; Family; Fc domain; Fibrin; Foundations; Future; Generations; Goals; Hemorrhage; High Pressure Liquid Chromatography; Human; Hydrolysis; Hypersensitivity; Hypotension; Immunization; Immunotherapy; In Vitro; Individual; Inflammation; Inflammatory; Kidney Failure; Knowledge; Lead; Libraries; Matrix Metalloproteinases; Medical; Metalloproteases; Monoclonal Antibodies; Necrosis; Pain; Peptide Hydrolases; Persons; Platelet Aggregation Inhibition; Preparation; Prevention; Production; Proteomics; Public Health; Reaction; Research; Research Project Grants; Risk; STEM research; Shock; Skin; Snake Bites; Snake Venoms; Snakes; Specificity; Structure; System; Testing; Therapeutic; Therapeutic Monoclonal Antibodies; Therapeutic Uses; Thrombocytopenia; Tissues; Toxin; Toxinology; Universities; Variant; Venoms; Viperidae; Work; World Health Organization; antibody libraries; antitoxin; base; cost; cytotoxic; design; efficacy evaluation; global health; health organization; human monoclonal antibodies; humanized monoclonal antibodies; in vivo; inhibitor; member; nanobodies; neglected tropical diseases; neutralizing antibody; next generation; novel; screening; sensor; side effect; systemic inflammatory response

PROJECT SUMMARY In this R21 award, we aim to develop a novel and broad neutralizing human monoclonal antibody for treating snakebite envenoming by rational structure-based design in order to produce a more effective and safer next generation antivenom. Snake envenomation is a serious global public health concern and ranked on the Wor ld Health Organization’s list of neglected tropical diseases, killing on average 125,000 people per year and leaving another 400,000 permanently disabled. The majority of snake envenomation in the US, inflicted by members of the snake family Viperidae, causes local tissue damage (such as myonecrosis, blisters, and local inflammation and pain) and systemic effects, including hemorrhage and coagulopathies which can lead to shock, renal failure and death. Snake venom metalloproteinases are major causative agents for spontaneous systemic bleeding and coagulopathies. Current antivenoms, produced by immunization of domestic animals, have limited efficacy in the prevention of both local and systemic effects of Viperidae envenomation as well as an associated risk of hypersensitivity reactions. Our long-term goal is to develop novel, effective humanized antivenom therapeutics for Viperidae envenomation. The objective of this project is to test the hypothesis that camelid-inspired inhibitory paratope synthetic human antibodies targeted to the active site of medically-relevant viperid venom metalloproteinases (svMPs) can provide broad antivenom protection without cross-reaction with human metalloproteinases and without the risk of hypersensitivity. This objective will be addressed through our established collaboration of complementary expertise between the snake venom toxinology team at National Natural Toxins Research Center (NNTRC) and the antibody discovery team at University of California Riverside (UCR). To test our hypothesis, we will address the following three Specific Aims. Aim 1: Qualitative and Quantitative Characterization of the hemorrhagic activity of viperid svMPs (Galan), Aim 2: Discovery of Broadly Neutralizing svMP-Specific Human mAbs (Ge). Aim 3: Evaluation of the antivenom efficacy of svMP inhibitory mAbs in vitro (Ge) and in vivo (Sanchez). The proposed research is significant because it will advance our understanding of the hemorrhagic aspects caused by snake envenomation at biochemical/cellular levels and develop effective humanized mAb antivenoms, which will be directly translatable for therapeutic use. The novelties of our project are (1) development and application of a novel Hemorrhage Score system to characterize svMPs; (2) isolation of humanized svMP-specific antivenom mAbs from libraries carrying novel convex paratopes; (3) development groundbreaking functional (rather than binding-based) HTS for facile discovery of mAbs inhibiting hemorrhagic snake toxins; and (4) potentially shifting the conventional antivenom production into specific neutralizing humanized mAb therapeutics.

项目总结 在这个R21奖项中，我们的目标是开发一种新的、广泛的中和人源单抗用于治疗 通过合理的结构设计毒蛇咬伤的毒液，以生产出更有效、更安全的NEXT 世代抗蛇毒血清。 蛇毒是一个严重的全球公共卫生问题，在世界排名中 LD 世界卫生组织的被忽视的热带病名单， 每年平均杀死12.5万人， 另有40万人永久残废。在美国，大多数蛇毒是由 蛇科的成员，会引起局部组织损伤(如肌坏死、水泡和局部 炎症和疼痛)和全身影响，包括出血和凝固性疾病，这可能导致 休克，肾功能衰竭和死亡。蛇毒金属蛋白酶是自发性的主要致病因素 全身出血和凝固性疾病。目前的抗蛇毒血清是通过家畜免疫产生的， 在预防毒蛇毒液的局部和全身影响以及 与过敏反应相关的风险。我们的长期目标是开发新颖、有效的人性化产品 毒蛇毒化的抗蛇毒治疗药物。这个项目的目标是检验这样一个假设 骆驼刺激的抑制性副表位合成的人抗体靶向医学相关的活性部位 蛇毒金属蛋白酶(Svmps)可以提供广泛的抗蛇毒保护作用，而不会与 人的金属蛋白酶，无过敏风险。这个目标将通过我们的 在国家毒物研究所蛇毒毒理学小组之间建立了互补的专业合作 加州大学天然毒素研究中心(NNTRC)和抗体发现小组 河滨大学(UCR)。为了验证我们的假设，我们将解决以下三个具体目标。目标1：定性 和Vperid svMPs(Galan)出血活性的定量表征，目标2：发现 广泛中和SVMP特异性人单抗(GE)。目的3：评价SVMP的抗蛇毒效果 体外抑制单抗(GE)和体内抑制单抗(Sanchez)。这项拟议的研究意义重大，因为它将 从生化/细胞角度加深对蛇毒出血的认识 并开发有效的人源化mAb抗蛇毒血清，可直接翻译用于治疗用途。 我们项目的创新之处在于(1)开发和应用了一种新的出血评分系统 鉴定SVMP；(2)从含新基因的文库中分离人源化SVMP特异性抗蛇毒单抗 凸拟合法；(3)开发突破性的功能性(而不是基于结合的)HTS，以方便 发现抑制出血性蛇毒的单抗；以及(4)潜在地改变了传统的抗蛇毒血清 生产成特定的中和人源化单抗疗法。