Roles for Cbl and ESCRT Proteins in 5-HT Receptor Function

Cbl 和 ESCRT 蛋白在 5-HT 受体功能中的作用

• 批准号: 7684371
• 负责人: John R Raymond
• 金额: --
• 依托单位: RALPH H JOHNSON VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7684371
• 关键词: Acquired Immunodeficiency Syndrome; Address; Age; Aggressive behavior; Agonist; Anxiety; Asthma; Atherosclerosis; Award; Back; Binding; Biochemical; Biological Process; Blood Vessels; Cell Fractionation; Cell Proliferation; Cell Surface Proteins; Cell membrane; Cell surface; Cells; Cellular biology; Chimeric Proteins; Clinical; Complex; Confocal Microscopy; Cytoskeleton; Data; Diabetes Mellitus; Disease; Drug Delivery Systems; Electrolytes; Family; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; Gene Silencing; Growth Factor Receptors; HTR2A gene; Heart failure; Homeostasis; Hormonal; Hormones; Human; Human Genome; Hypertension; Immune System Diseases; Incidence; Intervention; Kidney Diseases; Lead; Length; Light; Liquid substance; Malignant Neoplasms; Mediating; Mediator of activation protein; Membrane; Mental Depression; Mental disorders; Methodology; Methods; Mission; Molecular; Moods; Nature; Neurodegenerative Disorders; Neurotransmitters; Odors; Patient Care; Peptides; Pharmaceutical Preparations; Physiology; Play; Population; Principal Investigator; Protein Family; Proteins; Publications; Recycling; Regulation; Research; Role; Serotonin; Severities; Sex Behavior; Signal Transduction; Sleep; Sleep Disorders; Sorting - Cell Movement; Source; Stimulus; Stroke; Taste Perception; Therapeutic; Transfection; Traumatic Brain Injury; Vascular Diseases; Vascular Headaches; Vesicle; Veterans; Wakefulness; Work; abstracting; base; body system; cell growth; clinically relevant; extracellular; fundamental research; hepatocyte growth factor-regulated tyrosine kinase substrate; immune function; insight; knowledge base; member; novel; novel therapeutics; programs; protein complex; protein transport; public health relevance; receptor; receptor function; receptor recycling; therapeutic target; trafficking

DESCRIPTION (provided by applicant): Project Summary/Abstract The objective of this proposal is to unravel novel molecular mechanisms of regulation of serotonin (5-HT) receptors, which are cell surface proteins that transduce extracellular signals into intracellular signals. 5-HT receptors are prototypical G protein-coupled receptors (GPCRs), constitute a significant portion of the human genome. They participate in nearly every aspect of physiology and in many disease states. Nevertheless, our understanding of how these receptors transfer signals remains incomplete. In particular, many aspects of their intracellular trafficking, and the specific proteins involved in determining the fate of GPCRs, remain mysterious. This will be accomplished using the following methodologies: confocal microscopy, subcellular fractionation, biochemical methods, fusion proteins and transfections into -- or silencing of genes expressed in -- Hek293 cells. The hypothesis upon which this work is based is that two key protein families known to be involved in degradation of growth factor receptors - Cbl and HRS - are critical mediators of recycling of 5-HT2A receptors. The first aim focuses on c-Cbl, and the second on HRS. The work is significant in that our preliminary data demonstrate unexpected roles for c-Cbl and HRS in recycling (rather than degradation) of 5-HT2A receptors. The work is clinically relevant in that 5-HT regulates mood, aggression, anxiety, sexual behavior, sleep-wakefulness, vascular tone, immune function, cell growth and proliferation, and fluid and electrolyte homeostasis. This work is particularly germane to the Veteran population in that those disorders are frequently encountered in VA clinical settings. Importantly, the incidence and severity of many of those maladies will worsen as our Veteran population continues to age. The topic of this proposal also is significant because of the fundamental importance of understanding GPCR signal transduction as it relates to all functions of human cells. GPCRs comprise the largest known family of integral membrane receptor proteins. GPCRs represent key therapeutic targets for the treatment of many diseases of Veterans including hypertension, atherosclerosis, stroke, heart failure, cancer, asthma, renal diseases, diabetes mellitus, traumatic brain injury, psychiatric diseases, neurodegenerative diseases, and immune disorders including AIDS. Ultimately, the type of fundamental research proposed in this application will likely lead to new therapeutic insights and strategies that will form the basis for clinical interventions in multiple disease states. PUBLIC HEALTH RELEVANCE: Relevance of the Research to the VA Patient Care Mission The work covered in this proposal is relevant to the VA patient care mission in two respects. First, it will add to our knowledge about how 5-HT regulates mood, aggression, anxiety, sexual behavior, sleep-wakefulness, vascular tone, immune function, cell growth and proliferation, and fluid and electrolyte homeostasis. This work is particularly germane to the Veteran population in that anxiety, sleep disorders, depression, vascular headache, atherosclerotic-thrombotic vascular diseases and hypertension are frequently encountered in VA clinical settings. Importantly, the incidence and severity of many of those maladies will worsen as our Veteran population continues to age. The second reason why the topic of this proposal is significant involves the fundamental importance of understanding GPCR signal transduction as it relates to all functions of human cells. GPCRs comprise the largest known family of integral membrane receptor proteins. Recent estimates suggest that the GPCRs represent H1,000 members, and the GPCR family makes up > 1% of the human genome. GPCRs respond to a wide variety of stimuli including hormones, neurotransmitters, light, odor, taste, and peptides. Consequently, they play critical roles in the regulation of all organ systems and hormonal functions. The central importance of GPCRs in human physiology and diseases is highlighted by the fact that more than half of all medications prescribed today are targeted at GPCRs. GPCRs represent key therapeutic targets for the treatment of many diseases of Veterans including hypertension, atherosclerosis, stroke, heart failure, cancer, asthma, renal diseases, diabetes mellitus, psychiatric diseases, neurodegenerative diseases, and immune disorders including AIDS. Ultimately, the type of fundamental research proposed in this application will likely lead to new therapeutic insights and strategies that will form the basis for clinical interventions in multiple disease states.

描述（由申请人提供）： 项目概要/摘要本提案的目的是揭示5-羟色胺（5-HT）受体调节的新分子机制，5-HT受体是将细胞外信号转换为细胞内信号的细胞表面蛋白质。5-HT受体是典型的G蛋白偶联受体（GPCR），构成人类基因组的重要部分。它们几乎参与生理学的各个方面和许多疾病状态。然而，我们对这些受体如何传递信号的理解仍然不完整。特别是，它们的细胞内运输的许多方面，以及参与决定GPCR命运的特定蛋白质，仍然是神秘的。这将使用以下方法完成：共聚焦显微镜、亚细胞分级分离、生物化学方法、融合蛋白和转染到Hek 293细胞中或沉默Hek 293细胞中表达的基因。这项工作所基于的假设是，已知参与生长因子受体降解的两个关键蛋白质家族- Cbl和HRS -是5-HT 2A受体再循环的关键介质。第一个目标集中在c-Cbl，第二个目标集中在HRS。这项工作是重要的，因为我们的初步数据表明c-Cbl和HRS在5-HT 2A受体的再循环（而不是降解）中具有意想不到的作用。这项工作与临床相关，因为5-HT调节情绪、攻击性、焦虑、性行为、睡眠-觉醒、血管张力、免疫功能、细胞生长和增殖以及液体和电解质稳态。这项工作是特别密切相关的退伍军人群体，这些疾病经常遇到的VA临床设置。重要的是，随着我们的退伍军人人口继续老龄化，许多这些疾病的发病率和严重程度将恶化。该提案的主题也很重要，因为理解GPCR信号转导的根本重要性，因为它涉及人类细胞的所有功能。GPCR包含已知最大的整合膜受体蛋白家族。GPCR代表用于治疗退伍军人的许多疾病的关键治疗靶标，所述疾病包括高血压、动脉粥样硬化、中风、心力衰竭、癌症、哮喘、肾病、糖尿病、创伤性脑损伤、精神疾病、神经变性疾病和免疫病症（包括AIDS）。最终，本申请中提出的基础研究类型可能会导致新的治疗见解和策略，这些见解和策略将成为多种疾病状态下临床干预的基础。 公共卫生相关性： 研究与VA患者护理使命的相关性本提案中涵盖的工作在两个方面与VA患者护理使命相关。首先，它将增加我们对5-HT如何调节情绪、攻击性、焦虑、性行为、睡眠-觉醒、血管张力、免疫功能、细胞生长和增殖以及液体和电解质稳态的认识。这项工作是特别密切的退伍军人人群中，焦虑，睡眠障碍，抑郁症，血管性头痛，动脉粥样硬化血栓性血管疾病和高血压经常遇到的VA临床设置。重要的是，随着我们的退伍军人人口继续老龄化，许多这些疾病的发病率和严重程度将恶化。本提案主题重要的第二个原因涉及理解GPCR信号转导的根本重要性，因为它涉及人类细胞的所有功能。GPCR包含已知最大的整合膜受体蛋白家族。最近的估计表明，GPCR代表H1,000成员，GPCR家族占人类基因组的1%以上。GPCR对各种各样的刺激做出反应，包括激素、神经递质、光、气味、味觉和肽。因此，它们在所有器官系统和激素功能的调节中发挥关键作用。GPCR在人类生理学和疾病中的核心重要性突出表现在以下事实：今天处方的所有药物中有一半以上是针对GPCR的。GPCR代表用于治疗退伍军人的许多疾病的关键治疗靶标，所述疾病包括高血压、动脉粥样硬化、中风、心力衰竭、癌症、哮喘、肾脏疾病、糖尿病、精神疾病、神经变性疾病和免疫病症（包括AIDS）。最终，本申请中提出的基础研究类型可能会导致新的治疗见解和策略，这些见解和策略将成为多种疾病状态下临床干预的基础。