Regulation and Function of Urocortins and their Receptor

尿皮质素及其受体的调节和功能

• 批准号: 9127637
• 负责人: Aditi Bhargava
• 金额: $ 34.09万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9127637
• 关键词: Address; Adrenal Glands; Adult; Adverse effects; Agonist; Amino Acids; Animal Disease Models; Animals; Anxiety; Atrophic; Behavior; Biological Factors; Blood; Blood Vessels; Brain; CRF receptor type 1; CRF receptor type 2; Cardiovascular system; Cell membrane; Cell physiology; Cells; Cellular Stress Response; Childhood; Clinical Trials; Co-Immunoprecipitations; Colitis; Corticotropin-Releasing Hormone; Corticotropin-Releasing Hormone Receptors; Coupled; Coupling; Crohn' s disease; Cues; Data; Diagnosis; Disease; Disease Outcome; Drug Targeting; Early Endosome; Endoplasmic Reticulum; Environment; Environmental Risk Factor; Etiology; Extravasation; Female; Flow Cytometry; Functional disorder; G Protein-Coupled Receptor Genes; GTP-Binding Proteins; Gastrointestinal Diseases; Gastrointestinal tract structure; Genetic; Genotype; Heat shock proteins; Hormones; Hypertrophy; ITGAM gene; Immune; Immune response; Immune system; Immunomodulators; Immunoprecipitation; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory disease of the intestine; Intervention; Intestines; Knock-out; Lead; Leadership; Ligands; MAPK14 gene; Mass Spectrum Analysis; Measures; Mediating; Membrane Microdomains; Mental Depression; Modeling; Molecular; Mood Disorders; Mus; Neuropeptides; Organ; Outcome; Pancreas; Pathogenesis; Pathology; Patients; Permeability; Phosphorylation; Phosphotransferases; Plasma; Police; Proteins; Regulation; Reporting; Research; Resolution; Role; Scaffolding Protein; Sex Characteristics; Signal Pathway; Signal Transduction; Site; Spleen; Stress; Stress and Coping; Symptoms; System; Testing; Thymus Gland; Ulcerative Colitis; United States; United States National Institutes of Health; acute pancreatitis; astressin; biological adaptation to stress; cost; disorder subtype; experience; hypothalamic-pituitary-adrenal axis; male; migration; mortality; neutrophil; novel; peptide hormone; preclinical study; psychological stressor; receptor; response; scaffold; sex; therapeutic target; trafficking; urocortin

SUMMARY Inflammatory bowel disease (IBD) is one of the most prevalent GI diseases in the United States, costing ~ $1.7 billion annually in treatment. The etiology of IBD remains unknown. Although IBD outcome is clearly influenced by interactions involving the brain, gut, immune system, and the environment, the contributing biological factors remain under-characterized and have been identified as a major gap in understanding the etiology of IBD. Our proposal directly addresses this gap by elucidating biological factors, such as the components of the corticotropin-releasing factors (CRF) system, whose modulation can directly influence the gut and the brain. In addition to the physical symptoms of IBD, patients often experience stress-related mood disorders. The CRF system mediates the classic stress response via activation of the hypothalamic-pituitary-adrenal (HPA) axis. However, the cellular mechanisms by which the components of the CRF system bring about their immunomodulatory and stress-coping actions during IBD remain unknown. Our preliminary data shows that the stress receptor CRF2 is a key determinant of cellular stress responses during an inflammatory insult. Our over- arching hypothesis is that CRF and urocortins, differentially signaling through CRF1 and CRF2, govern intestinal inflammation at the molecular, cellular and organ levels in a sex-specific manner. We propose to test our hypothesis in three specific aims. In Aim 1, we will determine the sex-specific role of the CRF2 receptor in modulating aspects of GI permeability, organ mass, and immune cell function at baseline and during colitis. Our pilot data shows that the gut is “leakier” in female than in male mice. This leakiness is abrogated in mice lacking the CRF2 receptor. By measuring vascular leak and immune cell changes by flow cytometry, we will determine whether CRF2 receptor deficiency alters immune cell recruitment or migration during colitis, locally at the site of inflammation, and in the blood and spleen. The role of kinases and their targets in manifesting sex-specific effects during IBD will be ascertained. Changes in the components of the CRF system in the brain during colitis will be correlated with changes in anxiety- and depression-like behaviors. In Aim 2, we will determine the sex-specific role of urocortins and CRF agonists in modulating aspects of GI permeability, organ mass, and immune cell function at baseline and during colitis. By using approaches defined in Aim 1, we will delineate the role of 4 different agonists for the CRF2 receptor in modulating of function of CRF2 in a sex-specific manner. In Aim 3, we will determine if heteromerization of CRF receptors (CRF1:CRF2) and/or their association with scaffold proteins and heat shock proteins results in differential trafficking and signaling by their four ligands. By making tagged versions of CRF receptors and using a combination of immunoprecipitation and mass spectrometry, we will identify ancillary proteins that interact with CRF receptors to mediate context-specific signaling and trafficking. This proposal will define sex-specific signaling pathways that can be targeted for therapeutic purposes for patients with IBD.

概括 炎症性肠病（IBD）是美国最普遍的胃肠道疾病之一，耗资约1.7美元 每年十亿次治疗。 IBD的病因仍然未知。尽管IBD结果显然受到影响 通过涉及大脑，肠道，免疫系统和环境的相互作用，促成生物学因素 保持特征不足，并被确定为理解IBD病因的主要差距。我们的 提案直接通过阐明生物学因素来解决这一差距，例如 皮质激素释放因子（CRF）系统，其调节可以直接影响肠道和大脑。 除了IBD的身体症状外，患者经常患有与压力有关的情绪障碍。 CRF 系统通过激活下丘脑 - 垂体 - 肾上腺（HPA）轴介导了经典的应力反应。 但是，CRF系统的组成部分带来的细胞机制 IBD期间的免疫调节和应力促进作用尚不清楚。我们的初步数据表明 应力受体CRF2是炎症损伤期间细胞应力反应的关键决定剂。我们的 拱形假设是，通过CRF1和CRF2发出不同信号的CRF和尿道素控制 以性别特异性方式在分子，细胞和器官水平上处于肠道炎症。我们建议 在三个特定目标中检验我们的假设。在AIM 1中，我们将确定CRF2的性别特定作用 基线时GI通透性，器官质量和免疫细胞功能的调节方面的受体 在结肠炎期间。我们的飞行员数据表明​​，女性的肠道比雄性小鼠“漏水”。这种泄漏是 在缺乏CRF2受体的小鼠中被actrog。通过测量血管泄漏和免疫细胞的变化 细胞仪，我们将确定CRF2受体缺乏症是否改变了免疫细胞募集或迁移 在结肠炎期间，在炎症部位，血液和脾脏。激酶及其的作用 将确定在IBD期间表现出性别特异性效应的目标。组件的变化 结肠炎期间大脑中的CRF系统将与动画和抑郁症状行为的变化有关。 在AIM 2中，我们将确定尿道素和CRF激动剂在调节方面的性别特异性作用 基线和结肠炎期间的GI通透性，器官质量和免疫细胞功能。通过使用 在AIM 1中定义的方法，我们将描述4种不同激动剂在CRF2受体中的作用 以性别特定方式调节CRF2的功能。在AIM 3中，我们将确定是否存在异构化 CRF受体（CRF1：CRF2）和/或它们与脚手架蛋白和热激蛋白的关联 导致四个配体的贩运差异和信号传导。通过制作CRF的标记版本 受体并结合免疫沉淀和质谱法，我们将确定辅助 与CRF受体相互作用以介导上下文特异性信号传导和运输的蛋白质。该提议将 定义性别特异性信号通路，可以针对IBD患者进行治疗目的。