Evaluation of Toxicity Score and Precise Snake Venom Analytics for Next Generation Antivenom Development

下一代抗蛇毒血清开发的毒性评分评估和精确蛇毒分析

• 批准号: 10394860
• 负责人: Jacob Anthony Galan
• 金额: $ 13.8万
• 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: 2020-05-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10394860
• 关键词: Address; Antibody Therapy; Antidotes; Antivenins; Biochemistry; Biological; Biological Markers; Biomedical Research; Blood Coagulation Disorders; Blood coagulation; Cells; Cessation of life; Chemistry; Child; Collection; Complex; Crotalid; Data; Development; Dose; Enzymes; Escherichia coli; Escherichia coli Proteins; Evaluation; Faculty; Foundations; Functional disorder; Future; Geographic Locations; Goals; Health; Hemorrhage; Human; Immune response; Individual; Injury; Knowledge; Laboratories; Light; Mass Spectrum Analysis; Medical; Metabolic; Metalloproteases; Methods; Modeling; Molecular; Mus; Necrosis; Pathology; Peptides; Plasma; Plasmids; Play; Procedures; Prognostic Marker; Proteins; Proteomics; Recombinants; Reporting; Research; Research Project Grants; Research Training; Role; STEM program; Schools; Science; Snake Bites; Snake Venoms; Snakes; Structure; Students; Technology; Testing; Texas; Therapeutic; Time; Toxic effect; Toxin; Toxinology; Training; Universities; Venoms; Woman; acute toxicity; base; cDNA Library; career; design; diagnostic biomarker; exosome; experimental study; global health; graduate student; in vivo; innovation; insight; interest; member; men; method development; mouse model; neglected tropical diseases; new technology; next generation; novel; programs; protein expression; protein purification; statistics; stem; tool; tool development; treatment strategy; undergraduate student

ABSTRACT The overall goal of this project is to engage undergraduate and graduate students in innovative studies in molecular toxinology that have the potential to contribute to important advances in our understanding of snake venom pathology and the development of novel antibody-based therapeutics. According to WHO statistics, it is estimated that in 2020 there will be 125,000 new deaths world-wide reported due to snake envenomation. In spite of its importance for snake bite envenomation as a neglected tropical disease and global health issue, very little is known of the mechanism of action of snake venom and intrinsic toxic/lethal contribution. The scientific foundation for our study is that a specific combination of snake venom toxins plays a critical role in snakebites by causing necrosis, coagulopathy, severe hemorrhaging and eventually death. Inasmuch as individual toxins have been evaluated, synergistic mechanism for snake venom pathology are lacking and warranted for antidote development. We hypothesize that precise toxicity evaluation and snake venom analytics of crude/fractionated venom and recombinant toxins will provide a model in which priority can be given to the most toxic venom component(s). To test our hypothesis, we will address (3) Specific Aims: Specific Aim 1: To profile snake venom composition, combinational assessment and intrinsic toxicity. We will purify and characterize snake venom of Crotalid snake species. Characterization will include profiling snake venom to identify the toxins in each fraction and abundance. Students will investigate the mechanism of action different combinations of venom fractions. We will characterize the lethal concentrations/doses of each venom fraction on HUVEC cells, blood coagulation, and will validate the toxicity score for each fraction using an in vivo lethality mouse model. Specific Aim 2: To produce recombinant venom peptides. We will clone these toxins from our collection of cDNA libraries and will generate plasmids and transform competent E.coli cells for protein expression and purification. Development of methods to produce recombinant toxins is essential for antivenom development and investigating toxicity and targets for medically relevant venom peptide. These studies will also be essential to use toxins in solving structures to venom toxins or structures in complexes with known targets. Specific Aim 3: To identify snake venom biomarkers in plasma exosomes. We will explore the “venom-reactome” using novel Evtrap technology and proteomics in discovery-based analysis of snake venom biomarkers from mouse plasma exosomes. Comprehensive snake venom biomarker analysis will shed new light on the pathophysiology, metabolic, biological, cellular and immunological responses to snake venom.

抽象的 该项目的总体目标是让本科和研究生从事创新研究 分子毒素学有潜力为我们的蛇理解做出重要进展 毒液病理学和基于新型抗体治疗的发展。根据谁的统计数据，这是 估计在2020年，全世界将有125,000例新死亡因蛇的影响。在 它对蛇咬入的重要性是一种被忽视的热带疾病和全球健康问题， 关于蛇毒和内在的有毒/致命贡献的作用机理，知之甚少。 我们研究的科学基础是，蛇毒毒素的特定组合在 通过引起坏死，凝血病，严重的出血和最终死亡而导致蛇位。恰当 已经评估了个体毒素，缺乏蛇毒病理学的协同机制 有必要用于解毒剂开发。我们假设精确的毒性评估和蛇毒 原油/分级毒液和重组毒素的分析将提供优先级的模型 可以给予最有毒的毒液成分。为了检验我们的假设，我们将解决（3）特定的 目的：特定目的1：介绍蛇毒组成，结合评估和内在的 毒性。我们将净化并表征crotalid蛇种的蛇毒。表征将包括 分析蛇毒液以鉴定每个馏分和抽象中的毒素。学生将调查 作用机理的毒液分数的不同组合。我们将描述致命 每个毒液的浓度/剂量在HUVEC细胞上，血液凝结，并将验证毒性 使用体内致死性鼠标模型为每个分数分数。特定目标2：产生重组 毒液肽。我们将从我们的cDNA库中克隆这些毒素，并产生质粒 并转化竞争性大肠杆菌细胞以进行蛋白质表达和纯化。开发方法 产生重组毒素对于抗蛇毒的发展和研究毒性和目标至关重要 医学相关的毒胡椒。这些研究也将对于将毒素求解结构进行 具有已知靶标的复合物中的毒素毒素或结构。特定目标3：识别蛇毒 血浆外泌体中的生物标志物。我们将使用新颖的Evtrap技术探索“毒液 - 复反应仪”和 蛋白质组学基于发现小鼠血浆外泌体的蛇毒生物标志物的蛋白质组学。 全面的蛇毒生物标志物分析将为病理生理，代谢， 生物学，细胞和免疫学对蛇毒的反应。

项目成果

Examination of the Efficacy and Cross-Reactivity of a Novel Polyclonal Antibody Targeting the Disintegrin Domain in SVMPs to Neutralize Snake Venom.

• DOI: 10.3390/toxins13040254
• 发表时间: 2021-03-31
• 期刊: Toxins
• 影响因子: 4.2
• 作者: Szteiter SS;Diego IN;Ortegon J;Salinas EM;Cirilo A;Reyes A;Sanchez O;Suntravat M;Salazar E;Sánchez EE;Galan JA
• 通讯作者: Galan JA