The function(s) of serpin-2 in mosquito immunity and physiology

Serpin-2 在蚊子免疫和生理学中的功能

• 批准号: 8259121
• 负责人: Kristin Michel
• 金额: $ 36.98万
• 依托单位: KANSAS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8259121
• 关键词: Adult; Advanced Development; Affect; African; Aftercare; Anopheles gambiae; Arthropods; Binding; Biochemical; Biochemical Genetics; Biochemistry; Biological; Cessation of life; Cleaved cell; Clinical; Clip; Complex; Crystallography; Culicidae; Data; Development; Double-Stranded RNA; Emerging Communicable Diseases; Female; Generations; Goals; Health; Hemolymph; Human; Immune; Immune response; Immune system; Immunity; Injection of therapeutic agent; Insecta; Insecticide Resistance; Insecticides; Knowledge; Laboratories; Lead; Life Tables; Longevity; Malaria; Measures; Methodology; Microfluidics; Mission; Modeling; Molecular; Molecular Target; Natural Immunity; Nature; Parasites; Peptide Hydrolases; Pharmaceutical Preparations; Phenotype; Physiological; Physiology; Population; Populations at Risk; Prevention; Proteins; Proxy; Public Health; Research; Resistance; Resistance development; Serine Protease; Serine Proteinase Inhibitors; Serpins; Site; Structure; Techniques; Testing; Time; United States National Institutes of Health; Vector-transmitted infectious disease; base; design; disease transmission; disorder control; extracellular; in vivo; inhibitor/antagonist; innovation; killings; mathematical model; mortality; novel; pathogen; pressure; prevent; pro-phenoloxidase; programs; public health relevance; reproductive; resistance mechanism; serpin-2; small molecule; success; transmission process; tumor; vector; vector control

DESCRIPTION (provided by applicant): Malaria control and eradication strategies continue to primarily rely on vector control that is stymied by insecticide resistance in vector populations. In addition to resistance management, novel insecticides with new modes of action - particularly for which efficacy is not affected by existing resistance mechanisms - are required. The long-term goal is to develop a malaria vector control methodology based on a new insecticide target, the serine protease inhibitor serpin (SRPN)2 from the African malaria mosquito, Anopheles gambiae s.s.. SRPN2 is a key negative regulator in the extracellular proteinase cascade that controls activation of prophenoloxidase (proPO) and thus melanization - a powerful, arthropod-specific innate immune response. SRPN2 depletion from the hemolymph of adult female mosquitoes significantly reduces longevity with escalating daily mortality nine days after treatment. The objectives in this application are to identify the interactions of SRPN2 with its proteinase targets and to evaluate the malaria control potential of a SRPN2 inhibitor. The rationale for the proposed research is that detailed information on SRPN2's biological proteinase targets and mode of action will ultimately allow the design of small molecule inhibitors of SRPN2 that could act as an insecticide for mosquito vector control. Guided by our preliminary data, the following three specific aims will be pursued: (1) Determine the molecular targets of SRPN2 inhibitory function; (2) Determine the molecular interface between SRPN2 and its target proteinase(s); and (3) Model the impact of SRPN2 depletion on disease transmission. Under the first aim, a combination of biochemical and genetic approaches that have been used successfully in the applicant's laboratory will be used to identify serine proteinase targets of SRPN2 based on their ability to (a) form covalent complexes with the serpin and (b) revert the serpin's depletion phenotype. Under the second aim, the molecular interactions between SRPN2 and one of its target proteinases, CLIPB9, will be identified by protein crystallography and mutational analysis. Under the third aim, the potential effect of SRPN2 inhibition on malaria transmission and resistance development against a potential SRPN2 inhibitor will be assessed by mathematical modeling. The proposed research is innovative, as it will for the first time evaluate the mosquito immune system as a physiological target for novel insecticides. Additionally, this project will use microfluidics as a highly innovative approach to the study of mosquito innate immunity, which if successful will be transformative to the field of insect biochemistry. This project is significant as it will provide fundamental knowledge of the nature of serine proteinase cascades that regulate melanization in mosquitoes. Ultimately, it has the potential to advance the development of new a generation of insecticide targets for malaria control. PUBLIC HEALTH RELEVANCE: Vector-borne diseases, especially malaria, continue to be a major public health threat world-wide, with roughly half of the world population at risk of malaria. The proposed project is relevant to public health because the identification of novel insecticide targets in mosquitoes will allow the development of new vector control measures for malaria control. Therefore, the proposed project is relevant to the NIH-NIAID's mission in relation to the understanding and prevention of re-emerging infectious diseases.

描述（由申请人提供）：疟疾控制和根除策略继续主要依赖于载体种群中杀虫剂阻碍的载体控制。除了抵抗管理外，还需要采用新的作用模式的新型杀虫剂，尤其是对这种杀虫剂不受现有阻力机制的影响。长期目标是基于新的杀虫剂靶标，丝氨酸蛋白酶抑制剂SERPIN（SRPN）2从非洲疟疾蚊子，Anopheles gambiae S.S. SRPN2开发出一种疟疾媒介控制方法。SRPN2是一个关键的负调节剂，从而控制了外细胞蛋白酶壳的蛋白酶激活（Propans），因此（srpn）是一个关键的负调节剂，因此（旋转蛋白酶）激活了（Prop）的激活（Promoxase）。节肢动物特异性的先天免疫反应。成年雌性蚊子血淋巴的SRPN2耗竭可显着降低寿命，而每天在治疗后每天死亡率升级。本应用程序中的目标是确定SRPN2与其蛋白酶靶标的相互作用，并评估SRPN2抑制剂的疟疾控制潜力。拟议的研究的基本原理是，有关SRPN2生物蛋白酶靶标和作用方式的详细信息最终将允许设计SRPN2的小分子抑制剂，这些小分子抑制剂可以用作蚊子载体控制的杀虫剂。在我们的初步数据的指导下，将追求以下三个特定目标：（1）确定SRPN2抑制功能的分子靶标； （2）确定SRPN2及其靶蛋白酶之间的分子界面； （3）建模SRPN2耗竭对疾病传播的影响。在第一个目的下，在申请人实验室成功使用的生化方法和遗传方法的结合将用于识别SRPN2的丝氨酸蛋白酶靶标，其基于其（A）与SERPIN和（B）恢复Serpin的DEPLETETION的现象的能力。在第二个目标下，将通过蛋白质晶体学和突变分析来鉴定SRPN2和其靶蛋白酶之一ClipB9之间的分子相互作用。在第三个目标下，将通过数学建模评估SRPN2抑制对潜在SRPN2抑制剂的疟疾传播和抵抗发展的潜在影响。拟议的研究具有创新性，因为它将首次评估蚊子免疫系统作为新型杀虫剂的生理靶标。此外，该项目将使用微流体作为一种高度创新的方法来研究蚊子先天免疫，如果成功的话，它将对昆虫生物化学领域进行转化。该项目非常重要，因为它将提供有关调节蚊子中黑素化的丝氨酸蛋白酶级联体性质的基本知识。最终，它有可能促进新产生的疟疾控制杀虫剂靶标的发展。 公共卫生相关性：媒介传播的疾病，尤其是疟疾疾病，在全球范围内仍然是一个主要的公共卫生威胁，大约有一半的世界人口面临疟疾风险。拟议的项目与公共卫生有关，因为蚊子中新型杀虫剂靶标的鉴定将允许制定疟疾控制的新载体控制措施。因此，拟议的项目与NIH-NIAID有关的使命与理解和预防再出现传染病有关。