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.

项目概要/摘要 昆虫（包括蚊子等人类疾病媒介）的发育和繁殖 主要由两种亲脂性激素控制：蜕皮激素和保幼激素（JH）。虽然这些荷尔蒙 需要进入其靶细胞以发挥其生物效应，但对于分子方面几乎一无所知 调节激素跨细胞膜运输的机制。这是由于盛行的教条 在内分泌学中，亲脂性激素通过跨脂质双层的简单扩散进入和离开细胞。然而， 尽管有这一主导假设，但亲脂性激素运输的简单扩散模型并未得到支持 任何生物体中任何确凿的证据。事实上，最近的研究表明，一些亲脂性 激素，包括昆虫类固醇激素蜕皮激素，需要膜转运蛋白才能移动 穿过质膜。该项目的总体目标是识别和表征膜 JH 跨细胞膜运输所需的转运蛋白，从而挑战传统的 亲脂性激素通过简单扩散自由进入和离开细胞的范例。该方法将结合 初步研究中发现的表征 JH 转运蛋白 (JHT) 的体外和体内方法 使用果蝇模型系统。在目标 1 中，JHT 直向同源物在黄热病伊蚊中的功能 埃及伊蚊是寨卡病毒、黄热病病毒、基孔肯雅病毒和登革热病毒的主要传播媒介，将被彻底消灭。 使用分子遗传工具库进行体外研究。在目标2中，将进一步研究JHT函数 伊蚊的遗传。由于 JH 控制伊蚊和其他蚊子的生长和繁殖， 伊蚊 JHT 的表征有望帮助我们抗击这些最致命的疾病媒介 人类。事实上，在目标 3 中，将进行体外化学筛选，以确定能够抑制 伊蚊 JHT 的功能及其效果将在体内进行测试。因此，这个项目的意义并不在于 只是为了推翻内分泌学中长期存在的教条，同时也提供了一个关键的概念证明 作为种子化合物，用于开发控制蚊子和其他致命药物的新型药理学工具 病媒昆虫。