Juvenile hormone transporters in disease vector physiology

疾病媒介生理学中的保幼激素转运蛋白

• 批准号: 10658269
• 负责人: Naoki Yamanaka
• 金额: $ 37.96万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-24 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10658269
• 关键词: Adult; Aedes; Arboviruses; Biological; Biological Assay; Biological Models; Biology; CRISPR/Cas technology; Carrier Proteins; Cell Line; Cell membrane; Cells; Cellular Membrane; Chemicals; Chikungunya virus; Circulation; Culicidae; Cultured Cells; Cytoplasm; Data; Dengue Virus; Development; Diffuse; Diffusion; Disease Vectors; Double-Stranded RNA; Drosophila genus; Drosophila melanogaster; Ecdysone; Endocrinology; Female; Genetic Techniques; Geographic Distribution; Goals; Growth; Hormone Receptor; Hormones; Human; Hydrophobicity; In Vitro; Injections; Insecta; Juvenile Hormones; Lipid Bilayers; Liposomes; Luciferases; Mass Fragmentography; Mediating; Membrane; Membrane Transport Proteins; Methodology; Modeling; Molecular; Molecular Genetics; Mosquito Control; Mutagenesis; Mutation; Names; Organism; Orthologous Gene; Peripheral; Phenocopy; Phenotype; Physiological; Physiology; Proteins; Quantitative Reverse Transcriptase PCR; Reagent; Reporter; Reproduction; Role; Techniques; Terpenes; Textbooks; Thyroid Hormones; Travel; Work; Yellow Fever; ZIKA; combat; disorder control; global health; hormone deficiency; human disease; in vivo; in vivo evaluation; inhibitor; insect disease vector; knock-down; lipophilicity; loss of function; mosquito reproduction; mutant; novel; pharmacologic; receptor; screening; solute; steroid hormone; tool; trafficking; transcriptome sequencing; vector

PROJECT SUMMARY/ABSTRACT Development and reproduction of insects, including human disease vectors such as mosquitoes, are mainly controlled by two lipophilic hormones: ecdysone and juvenile hormone (JH). Although these hormones need to enter their target cells to exert their biological effects, almost nothing is known regarding molecular mechanisms that regulate hormone transport across cellular membranes. This is due to the prevailing dogma in endocrinology that lipophilic hormones enter and exit cells by simple diffusion across lipid bilayers. However, despite this dominant assumption, the simple diffusion model of lipophilic hormone transport is not supported by any conclusive evidence in any organism. Indeed, recent studies now suggest that some lipophilic hormones, including the insect steroid hormone ecdysone, require membrane transporter proteins to travel across plasma membranes. The overall objective of this project is to identify and characterize membrane transporters required for JH trafficking across cellular membranes, and to thereby challenge the conventional paradigm that lipophilic hormones freely enter and exit cells by simple diffusion. The approach will combine in vitro and in vivo approaches to characterize JH Transporter (JHT), which was discovered in preliminary studies using the fruit fly model system. In Aim 1, functions of the JHT ortholog in the yellow fever mosquito Aedes aegypti, the primary vector for Zika, yellow fever, chikungunya, and dengue viruses, will be thoroughly investigated in vitro using an arsenal of molecular genetic tools. In Aim 2, JHT functions will be further studied genetically in Aedes. As JH controls both growth and reproduction in Aedes and other mosquitoes, characterization of Aedes JHT is expected to aid our effort to combat these deadliest disease vectors for humans. Indeed, in Aim 3, in vitro chemical screening will be conducted to identify compounds that can inhibit functions of Aedes JHT, and their effects will be tested in vivo. The significance of this project is therefore not just to overturn the long-standing dogma in endocrinology, but also to provide a critical proof of concept as well as seed compounds for developing novel pharmacological tools to control mosquitoes and other deadly disease vector insects.

项目摘要/摘要 昆虫的开发和繁殖，包括人类疾病媒介，例如蚊子，是 主要由两种亲脂激素控制：ecdysone和少年激素（JH）。虽然这些激素 需要输入其靶细胞以发挥其生物学作用，几乎一无所知 调节激素跨细胞膜的机制。这是由于盛行的教条 在内分泌学中，亲脂激素通过简单扩散脂质双层进入和退出细胞。然而， 尽管存在这种主要假设，但不支持亲脂激素转运的简单扩散模型 通过任何生物体中的任何确定证据。确实，最近的研究现在表明一些亲脂性 激素，包括昆虫类固醇激素ecdysone，需要膜转运蛋白才能传播 跨质膜。该项目的总体目的是识别和表征膜 JH贩运跨细胞膜所需的转运蛋白，从而挑战常规 通过简单扩散自由进入和退出细胞的范式。该方法将结合在一起 体外和体内方法的表征JH转运蛋白（JHT），这是在初步研究中发现的 使用果蝇模型系统。在AIM 1中，JHT直系同源物在黄热病蚊子中的功能 Aegypti是Zika，黄热病，Chikungunya和登革热病毒的主要向量 使用分子遗传工具的武库在体外研究。在AIM 2中，将进一步研究JHT功能 遗传在艾德斯中。随着JH控制艾德河和其他蚊子的生长和繁殖， 预计艾德斯JHT的表征将有助于我们与这些最致命的疾病向量作斗争的努力 人类。实际上，在AIM 3中，将进行体外化学筛查，以鉴定可以抑制的化合物 AEDES JHT的功能及其效果将在体内进行测试。因此，该项目的意义不是 只是为了推翻内分泌学中长期存在的教条，但也提供了关键的概念证明 作为开发新型药理学工具来控制蚊子和其他致命的种子化合物 疾病媒介昆虫。