Neurotrophins in the Lung

肺中的神经营养素

• 批准号: 7792333
• 负责人: Y. S. Prakash
• 金额: $ 37.78万
• 依托单位: MAYO CLINIC ROCHESTER
• 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/7792333
• 关键词: Address; Affinity; Agonist; Area; Asthma; Biochemistry; Brain-Derived Neurotrophic Factor; Bronchoconstrictor Agents; Cells; Chronic Bronchitis; Cyclic ADP-Ribose; Data; Designer Drugs; Development; Disease; Family; Fluorescence; Foundations; Functional disorder; Generations; Goals; Growth Factor; Human; Hypersensitivity; Image; Immune; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Interleukins; Knock-in Mouse; Knowledge; Literature; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Measurement; Mechanics; Mediating; Molecular Biology; Morphology; Mus; Muscle Contraction; Myosin Light Chains; Myosin Regulatory Light Chains; NGFR Protein; Nerve; Nerve Growth Factor Receptors; Nerve Growth Factors; Nervous system structure; Neurons; Neurotrophic Tyrosine Kinase Receptor Type 2; Neurotrophin 3; Ovalbumin; Pathway interactions; Pharmacology; Phospholipase C; Physiology; Publishing; Pulmonary Emphysema; Receptor Activation; Regulation; Relative (related person); Research; Resistance; Rho-associated kinase; Role; Ryanodine Receptor Calcium Release Channel; Ryanodine Receptors; Sarcoplasmic Reticulum; Second Messenger Systems; Shortness of Breath; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Source; Techniques; Tissues; Transgenic Mice; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Up-Regulation; Work; airway hyperresponsiveness; airway inflammation; base; cytokine; immunocytochemistry; mouse model; neurotrophic factor; new therapeutic target; novel; public health relevance; receptor; receptor expression; respiratory smooth muscle; response; second messenger; Address; Affinity; Agonist; Area; Asthma; Biochemistry; Brain-Derived Neurotrophic Factor; Bronchoconstrictor Agents; Cells; Chronic Bronchitis; Cyclic ADP-Ribose; Data; Designer Drugs; Development; Disease; Family; Fluorescence; Foundations; Functional disorder; Generations; Goals; Growth Factor; Human; Hypersensitivity; Image; Immune; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Interleukins; Knock-in Mouse; Knowledge; Literature; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Measurement; Mechanics; Mediating; Molecular Biology; Morphology; Mus; Muscle Contraction; Myosin Light Chains; Myosin Regulatory Light Chains; NGFR Protein; Nerve; Nerve Growth Factor Receptors; Nerve Growth Factors; Nervous system structure; Neurons; Neurotrophic Tyrosine Kinase Receptor Type 2; Neurotrophin 3; Ovalbumin; Pathway interactions; Pharmacology; Phospholipase C; Physiology; Publishing; Pulmonary Emphysema; Receptor Activation; Regulation; Relative (related person); Research; Resistance; Rho-associated kinase; Role; Ryanodine Receptor Calcium Release Channel; Ryanodine Receptors; Sarcoplasmic Reticulum; Second Messenger Systems; Shortness of Breath; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Source; Techniques; Tissues; Transgenic Mice; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Up-Regulation; Work; airway hyperresponsiveness; airway inflammation; base; cytokine; immunocytochemistry; mouse model; neurotrophic factor; new therapeutic target; novel; public health relevance; receptor; receptor expression; respiratory smooth muscle; response; second messenger

DESCRIPTION (provided by applicant): An exciting, new investigative theme in airway physiology and pathophysiology is neurotrophins (NTs): growth factors including brain-derived neurotrophic factor (BDNF) known for their diverse roles in the nervous system. NTs and their receptors have now been found in different lung components including airway smooth muscle (ASM), with altered expression observed in asthma, allergy, and even lung cancer. While NTs may be derived from several sources, our published and preliminary data suggest that ASM is a target of NTs, and that NTs contribute not only to ASM contractility under normal circumstances, but also to increased contractility with airway inflammation (such as that induced by TNFa). The long term goal of the proposed studies is to understand the role of NTs in ASM physiology and pathophysiology. The overall hypothesis is that NTs enhance a) sarcoplasmic reticulum (SR) Ca2+ release and Ca2+ influx; and b) Ca2+ sensitivity for force generation in ASM. We propose that BDNF is a key NT influencing ASM contractility. Finally, airway inflammation enhances BDNF signaling, leading to an overall enhancement of [Ca2+]i and force. In this proposal, we will use human ASM and the ovalbumin (OVA) mouse model to examine the relative role of the BDNF receptors (high affinity TrkB vs. low affinity p75NTR) vis-¿vis ASM contractility. We hypothesize that TrkB is more important for [Ca2+]i regulation, while p75NTR regulates force. Using biochemistry, pharmacology, molecular biology, immunocytochemistry, fluorescence Ca2+ imaging, force measurement techniques, and lung mechanics, we will focus on specific mechanisms that may be regulated by BDNF: the second messengers IP3 (via phospholipase C PLC) and cyclic ADP ribose (via CD38) (Aim 1); SR Ca2+ release (IP3 receptor vs. ryanodine receptor (RyR) channels) (Aim 2); Ca2+ influx via store-operated Ca2+ entry (SOCE) (Aim 3) and the force regulatory mechanisms myosin light chain (MLC20) and rhoA/rho-kinase (Aim 4). These in vitro studies in human ASM will be integrated into the OVA mouse model applied in focus studies to the TrkB knockin mouse (where TrkB functionality is reversibly inhibited). We will explore the idea that inflammation induced by TNFa increases constitutive BDNF receptor expression, and alters specific [Ca2+]i and force regulatory mechanisms, thus priming ASM for enhanced response to both BDNF and bronchoconstrictor. The Specific Aims are: Aim 1: To determine mechanisms by which BDNF modulates second messenger signaling in human ASM; Aim 2: To determine mechanisms by which BDNF modulates SR Ca2+ regulation in human ASM; Aim 3: To determine mechanisms by which BDNF modulates SOCE in human ASM; Aim 4: To determine mechanisms by which BDNF modulates force regulation in human ASM; Aim 5: To determine the role of BDNF in ASM contractility in a mouse model of airway inflammation and hyperresponsiveness. PUBLIC HEALTH RELEVANCE. There is increasing recognition that abnormalities in airway smooth muscle contractility (exacerbated by inflammation) contribute to exaggerated airway narrowing and accompanying shortness of breath in clinically important diseases such as asthma and chronic bronchitis. In this regard, the potential role of growth factors called neurotrophins in regulation of airway contractility is an exciting and emerging area of research. By establishing the role of neurotrophins in airway narrowing with or without inflammation, the proposed studies will the foundation for better understanding of airway diseases, and potential development of new therapeutic targets.

描述（由适用提供）：气道生理和病理生理学中令人兴奋的新调查主题是神经营养蛋白（NTS）：包括脑衍生的神经营养因子（BDNF）在内的生长因素，以其在神经系统中的发散作用而闻名。现在已经在包括气道平滑肌（ASM）在内的不同肺部成分中发现了NTS及其受体，并且在哮喘，过敏甚至肺癌中观察到表达改变。尽管NTS可能来自多种来源，但我们已发布和初步数据表明ASM是NTS的目标，并且NTS不仅在正常情况下会贡献ASM的收缩性，而且还增加了使用气道注入（例如TNFA诱导的）。拟议的研究的长期目标是了解NT在ASM生理学和病理生理学中的作用。总体假设是NTS增强了A）肌质网（SR）Ca2+释放和Ca2+影响； b）在ASM中产生力的Ca2+灵敏度。我们建议BDNF是ASM的关键影响ASM收缩力。最后，气道注入增强了BDNF信号传导，从而导致[Ca2+] I和力的总体增强。在此提案中，我们将使用人类ASM和卵形蛋白（OVA）小鼠模型来检查BDNF受体（高亲和力TRKB与低亲和力P75NTR）的相对作用。我们假设TRKB对于[Ca2+] I调节更为重要，而P75NTR调节力。使用生物化学，药理学，分子生物学，免疫细胞化学，荧光CA2+成像，力测量技术和肺机制，我们将重点介绍可能受BDNF的特定机制：第二个Messenger ip3（通过磷脂酶C PLC）和环状ADP肋骨（通过CD38）（通过CD38）（AIM）（AIM） SR Ca2+释放（IP3受体与Ryanodine受体（RYR）通道）（AIM 2）； CA2+通过商店经营的Ca2+进入（SOCE）（AIM 3）和力调节机制肌球蛋白轻链（MLC20）和RhoA/Rho-kinase（AIM 4）的影响。这些在人ASM中的体外研究将集成到焦点研究中的OVA小鼠模型中（其中TRKB功能可逆抑制）。我们将探讨这样一种观念，即TNFA诱导的炎症会增加组成型BDNF受体的表达，并改变特定的[Ca2+] I和力调节机制，从而启动ASM，以增强对BDNF和BRONCHOCONTICTOR的反应。具体目的是：目标1：确定BDNF调节人类ASM中第二信使信号传导的机制；目标2：确定BDNF调节人ASM中SR Ca2+调控的机制；目标3：确定BDNF调节人类ASM中SOCE的机制；目标4：确定BDNF调节人ASM的力调节的机制； AIM 5：确定BDNF在ASM收缩性中的作用在气道注入和反应性过高的小鼠模型中。公共卫生相关性。人们越来越认识到气道平滑肌收缩力异常（炎症加剧）有助于夸大气道变窄，并参与临床重要疾病（例如哮喘和慢性支气管炎）的呼吸急促。在这方面，称为神经营养蛋白在气道收缩力调节中的生长因子的潜在作用是一个令人兴奋且新兴的研究领域。通过确定神经营养蛋白在气道狭窄或不发炎中的作用，提出的研究将基础更好地理解气道疾病，并潜在的新治疗靶标的发展。