Epigenetic regulation of hormone action in Tribolium and Aedes

谷盗和伊蚊激素作用的表观遗传调控

基本信息

批准号：
10417159
负责人：
SUBBA R PALLI
金额：
$ 29.87万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-02-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10417159
关键词：
Acetylation Adult Aedes Agonist Animals Antibodies Binding Bioinformatics Biological Assay Biological Metamorphosis Biological Models CRISPR/Cas technology Cell Culture Techniques Cells Complex Culicidae Data Deacetylation Development Developmental Process Disease Disease Vectors Ecdysone Ecdysteroids Ecdysterone Endocrinology Ensure Enzymes Epigenetic Process Female Food Genes Genome Goals Growth and Development function Histone Deacetylase Histone-Lysine N-Methyltransferase Histones Homologous Gene Hormone Receptor Hormones Human Insect Control Insect Hormones Insecta Insecticides Juvenile Hormones Knowledge Laboratories Life Ligands Literature Livestock Malignant Neoplasms Mass Spectrum Analysis Mediating Medicine Methods Methoprene Methylation Modeling Modification Molecular Pathway interactions Pest Control Physiological Physiological Processes Plants Play Principal Investigator Process Proteins Publishing RNA Interference Regulation Reporter Reproduction Reproductive Process Research Research Support Response Elements Role Steroid Receptors Testing Tissues Tribolium Vertebrates Western Blotting base chromatin immunoprecipitation demethylation ecdysone receptor enzyme activity epigenetic regulation experimental study gene product genome editing histone acetyltransferase histone demethylase histone modification hormone analog hormone regulation hormone response element human disease insect disease vector mass spectrometer programs promoter receptor receptor binding response success transcription factor transcriptome sequencing vector control yeast two hybrid system

项目摘要

PROJECT SUMMARY There has been a continuous demand for development of insect control methods that are target-specific. Juvenile hormone (JH) and ecdysteroids (20-hydroxyecdysone, 20E is the most active form) are the major hormones that regulate many physiological and developmental processes in insects. Since these hormones are not present in vertebrates, they represent attractive targets for development of insect control methods. Hindering this effort is the lack of understanding on the molecular basis of action of these hormones. There has been excellent recent progress in understanding the molecular basis of 20E and JH action. Key players, including receptors, co-activators, response elements and target genes involved in the action of these hormones have been identified. Non-steroidal ecdysone agonists and JH analogs have been discovered, and commercialized for the control of pests and disease vectors. Research supported by this program showed that epigenetic modifications, especially acetylation levels of histones regulated by histone acetyltransferases (HAT) and histone deacetylases (HDAC) modulate JH and 20E action. Hormone action requires multiple layers of regulation because the same circulating levels of hormones exert diverse functions in different tissues and under different physiological conditions. Based on the published literature and preliminary data included in this application, we hypothesize that epigenetic regulators HDACs, HATs, histone lysine methyltransferases (HKMT) and histone demethylases (HDM) modulate JH and 20E action in the regulation of postembryonic development and reproduction. The main goal of this proposal to test this hypothesis by studying the mechanisms of action of epigenetic modifiers in JH and 20E action using T. castaneum, TcA cells, Ae. aegypti and Aag2 cells as model systems. The two Specific Aims of the proposal are 1. To identify and characterize epigenetic modifiers required for 20E and JH regulation of postembryonic development and female reproduction and 2. To study mechanisms of action of epigenetic modifiers required for 20E and JH regulation of postembryonic development and female reproduction. In the first Specific Aim, RNAi, CRISPR/Cas9 genome editing methods, RNA sequencing, bioinformatics, RT-qPCR, enzyme assays and cell culture approaches will be used to identify and characterize HATs, HDACs, HKMTs and HDMs required for postembryonic development and female reproduction in T. castaneum and Ae. aegypti. In the second Specific Aim, the mechanisms of action of 10-12 epigenetic modifiers will be studied using western blots and ChIP, reporter, Two-hybrid and pull-down assays, and mass spectrometry approaches. Results from the proposed research will impact medicine by providing information for the development of vector control methods. This information could help in understanding epigenetics of human diseases such as cancer. The results from the proposed research may help to transform our understanding of endocrinology of disease vectors.

项目概要人们一直需要开发针对特定目标的昆虫控制方法。保幼激素（JH）和蜕皮类固醇（20-羟基蜕皮酮，20E 是最活跃的形式）是主要的调节昆虫许多生理和发育过程的激素。由于这些荷尔蒙虽然它们不存在于脊椎动物中，但它们代表了昆虫控制方法开发的有吸引力的目标。阻碍这一努力的是缺乏对这些激素作用的分子基础的了解。有最近在了解 20E 和 JH 作用的分子基础方面取得了出色的进展。关键球员，包括参与这些作用的受体、共激活剂、反应元件和靶基因激素已被鉴定。已发现非甾体蜕皮激素激动剂和 JH 类似物，并且商业化用于控制害虫和疾病媒介。该计划支持的研究表明表观遗传修饰，尤其是组蛋白乙酰转移酶调节的组蛋白乙酰化水平 (HAT) 和组蛋白脱乙酰酶 (HDAC) 调节 JH 和 20E 作用。激素作用需要多层因为相同的循环水平的激素在不同的组织中发挥不同的功能，在不同的生理条件下。根据已发表的文献和本报告中包含的初步数据应用中，我们假设表观遗传调节因子 HDAC、HAT、组蛋白赖氨酸甲基转移酶 (HKMT) 和组蛋白去甲基化酶 (HDM) 调节 JH 和 20E 在胚胎后调节中的作用发育和繁殖。该提案的主要目标是通过研究来检验这一假设表观遗传修饰剂在 JH 和 20E 作用中的作用机制，使用 T.castaneum、TcA 细胞、Ae。埃及伊蚊和 Aag2 细胞作为模型系统。该提案的两个具体目标是 1. 确定并描述胚胎后发育和雌性的 20E 和 JH 调节所需的表观遗传修饰剂 2. 研究 20E 和 JH 调节所需的表观遗传修饰剂的作用机制胚胎后发育和女性生殖。在第一个特定目标中，RNAi、CRISPR/Cas9 基因组编辑方法、RNA 测序、生物信息学、RT-qPCR、酶测定和细胞培养方法将用于识别和表征 HAT、HDAC、HKMT 和 HDM 所需的 T.castaneum 和 Ae 的胚胎后发育和雌性繁殖。埃及伊蚊。在第二个具体目的，将使用蛋白质印迹和 ChIP 研究 10-12 表观遗传修饰剂的作用机制，报告器、双杂交和下拉分析以及质谱方法。拟议的结果研究将通过为病媒控制方法的开发提供信息来影响医学。这信息可以帮助了解癌症等人类疾病的表观遗传学。结果来自拟议的研究可能有助于转变我们对疾病媒介内分泌学的理解。