Molecular mode of action of Juvenile hormone analogs

保幼激素类似物的分子作用模式

• 批准号: 10286849
• 负责人: SUBBA R PALLI
• 金额: $ 21.75万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10286849
• 关键词: Adolescent; Adult; Aedes; Affinity; Animals; Apoptosis; Autophagocytosis; Binding; Biological Assay; Biological Metamorphosis; CRISPR/Cas technology; Cells; Cessation of life; Code; Culicidae; Data; Development; Disease; Disease Vectors; Drosophila melanogaster; Dyes; Ecdysone; Electron Microscope; Ensure; Fleas; Fluorescence; Fluorescent Dyes; Genes; Genetic Transcription; Goals; Homologous Gene; Hormone Receptor; Human; Insecta; Insecticides; Juvenile Hormones; Knock-out; Knowledge; Larva; Life; Mediating; Medicine; Methods; Methoprene; Midgut; Molecular; Molting; Mosquito Control; Orthologous Gene; Phenotype; Plants; Proteins; Pupa; RNA; RNA Interference; Regulation; Reporter; Reporting; Research; Resistance; Resistance development; Salivary Glands; Stains; Testing; Tissues; Transgenic Organisms; Whiteflies; Work; XCL1 gene; Yellow Fever; activating transcription factor; analog; base; combat; experimental study; fly; genome editing; hormone analog; human pathogen; insect disease vector; knockout animal; mutant; novel; pathogen; preservation; prevent; response; screening; success; transcription factor; transgenic insect; transmission process; vector; vector control

Project Summary Adult mosquitoes transmit pathogens that cause deadly diseases in humans. Therefore, preventing adult emergence has been used as a strategy to block transmission of pathogens. Juvenile hormones (JH) prevent metamorphosis and adult emergence; juvenile hormone analogs (JHA) such as methoprene, hydroprene, and pyriproxyfen have been commercialized for controlling mosquitoes and other disease vectors. However, the molecular mode of action of JHAs in killing mosquitoes and other insects remains poorly understood. JHAs are thought to act by preventing larval-pupal metamorphosis. This is precisely how these compounds work in most lepidopteran insects. However, in dipteran insects such as the yellow fever mosquito, Aedes aegypti, JHA application does not prevent larval-pupal metamorphosis; the treated larvae undergo pupal ecdysis and die during the pupal stage. Preliminary studies showed that the pupae developed from methoprene treated larvae die due to the persistence of larval tissues. A transgenic CRISPR/Cas9 genome editing method was employed to knockout Ae. aegypti ortholog of the gene coding for ecdysone induced protein 93F (E93) which is known to regulate programmed cell death (PCD) of larval tissues. The phenotypes of Ae. aegypti E93 knockout animals are like that observed in JHA treated larvae, suggesting that JHA suppresses E93 expression in preventing the death of larval tissues, resulting in pupae containing both larval and pupal tissues leading to their death. This hypothesis will be tested by conducting experiments under two specific aims. In the first specific aim, larval tissues undergoing PCD will be examined to identify similarities and differences between JHA treatment and E93 knockout. The status of PCD in tissues dissected from JHA treated and E93 knockout larvae and pupae will be studied by staining with apoptosis and autophagy marker dyes. In the second specific aim, RNA and ATAC sequencing, RNAi and reporter assays will be employed to identify genes involved in larval tissue remodeling and mechanisms of regulation of their expression by JHA working through E93. The function of E93 acting as a pioneer and an activating transcription factor will be studied. Results from the proposed research will impact medicine by providing information for the development of vector control methods. The results from the proposed studies will answer long-standing questions on JHA action in mosquitoes and increase our understanding of JH action, which could help to develop novel, highly active, and safe methods to control mosquitoes and other vectors of pathogens that cause diseases in humans.

项目摘要 成年蚊子传播导致人类致命疾病的病原体。因此，防止成人 出现已被用作阻止病原体传播的策略。青少年激素（JH）预防 变形和成人出现；青少年激素类似物（JHA），例如甲丁烯，氢丁烯和 吡啶芬已被商业化用于控制蚊子和其他疾病媒介。但是， JHA在杀死蚊子和其他昆虫中的分子作用方式仍然很少了解。贾斯是 被认为是通过预防幼虫杆状变形而起作用的。这正是这些化合物在大多数的工作方式 鳞翅目昆虫。但是，在黄热病蚊子等双翅目昆虫中 应用不能预防幼虫型型变形；经过处理的幼虫经历了pupal骨的束缚并死亡 在Pupal阶段。初步研究表明，p pa是从甲酯处理的幼虫发展的 由于幼虫组织的持续存在而死亡。采用了转基因CRISPR/CAS9基因组编辑方法 淘汰AE。 ecdysone诱导蛋白93F（E93）的基因编码的埃及直系同源 调节幼虫组织的程序性细胞死亡（PCD）。 AE的表型。埃及E93淘汰动物 就像在JHA处理过的幼虫中观察到的那样，这表明JHA抑制了E93的表达 幼虫组织的死亡，导致pupa型含有幼虫和pup虫组织，导致其死亡。这 假设将通过在两个具体目标下进行实验来检验。在第一个特定目标中，幼虫 将检查正在接受PCD的组织，以确定JHA治疗和 E93淘汰赛。从经过JHA处理的组织和E93敲除幼虫和p的组织中的PCD状态 将通过凋亡和自噬标记染料染色来研究。在第二个特定目标中，RNA和 将使用ATAC测序，RNAi和记者测定法来鉴定涉及幼虫组织的基因 通过JHA通过E93进行的JHA对其表达调节的重塑和机制。功能 将研究充当先驱和激活转录因子的E93。提议的结果 研究将通过提供向向量控制方法的开发信息来影响医学。这 拟议的研究的结果将回答有关蚊子中JHA行动的长期问题 提高我们对JH行动的理解，这可能有助于开发新颖，高度活跃和安全的方法 控制蚊子和其他导致人类疾病的病原体载体。