Contribution of Toll-like Receptors in the Pulmonary Response to Nanoparticles an

Toll 样受体在肺部对纳米颗粒反应中的贡献

• 批准号: 8346247
• 负责人: Tara L Sabo-Attwood
• 金额: $ 32.64万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-15 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8346247
• 关键词: Address; Affect; Animal Model; Animals; Antibodies; Asbestos; Bacterial Infections; Behavior; Binding; Biological; Biological Assay; Biological Models; Breathing; Caliber; Cell physiology; Cells; Chemistry; Complex; Custom; Data; Development; Diesel Exhaust; Dimensions; Engineering; Epithelial Cells; Exposure to; Fiber; Functional disorder; Genetic; Goals; Health; Human; Image; Immune response; Immunity; In Vitro; Infection; Infectious Agent; Inflammation; Inflammatory; Influenza; Influenza A virus; Injury; Interferons; Invaded; Knockout Mice; Knowledge; Lead; Life; Lung; Lung diseases; Measures; Mediating; Medical; Mission; Modification; Molecular; Mus; NF-kappa B; Organ; Outcome; Particulate Matter; Pathway interactions; Predisposition; Process; Production; Property; Protein Family; Receptor Activation; Reporting; Role; Route; Screening procedure; Series; Signal Transduction; Structure of parenchyma of lung; Surface; System; Technology; Testing; Therapeutic; Tissues; Toll-Like Receptor Pathway; Toll-like receptors; Toxic effect; United States National Institutes of Health; Virus; Virus Diseases; Work; base; cytokine; cytotoxic; design; electronic structure; fluorescence imaging; immunoregulation; improved; in vitro Model; in vivo; inhibitor/antagonist; innovation; interest; mouse model; nanomaterials; nanoparticle; pandemic disease; particle; pathogen; receptor; receptor expression; response; safety practice; single walled carbon nanotube; synergism; transcription factor; ultrafine particle

DESCRIPTION (provided by applicant): Despite wide-scale use, we have limited understanding of the biotoxicity associated with nanoparticle (NP) exposures which has led to concerns regarding potential adverse health effects. Inhalation is likely a primary exposure route and the association between respired particulate matter with pulmonary disease, underscores the critical need to comprehend how NP impact the lungs. We have a particular interest in single-walled carbon nanotubes (SWNT) as they are widely used and possess a superficial resemblance to asbestos which may be relevant to their long term health consequences. Additional concerns surround the ability of NP to modulate the behavior of infectious agents. Increased susceptibility to infections as a result of NP exposure can have immense consequences particularly for viruses, such as influenza A (IAV), that are notorious for causing global pandemics. As realistic exposure scenarios are likely to involve multiple agents, triggering conserved signaling mechanisms may lead to enhanced detrimental responses that contribute to more severe health outcomes. Based on these notions, the overall objective of this application is to characterize the mechanisms controlling the primary immune response of lung epithelial cells exposed to SWNT and IAV singly and in combination. Our mechanistic focus will center on toll-like receptors (TLRs) as they are an early line of defense to foreign agents that enter in the body. We will test the hypothesis that SWNT of distinct chirality/diameter stimulate TLRs resulting in the production of pro-inflammatory cytokines through activation of the transcription factors NF-k¿ and IRFs. Furthermore, combined exposures of SWNT and IAV will synergistically activate TLR-driven pathways leading to enhanced inflammation and injury. This premise will be tested in 3 comprehensive specific aims which employ state-of-the-art technologies to (1) determine stimulation of distinct TLRs by SWNT having different chiral wrapping angles and diameters as well as by IAV, (2) assess direct interactions of SWNT and IAV and investigate the role of select TLR modulation by these agents in lung epithelial cells and (3) examine the lung inflammatory and clearance response of single and sequential exposures of SWNT and IAV mediated, in part, by TLR-NF-k¿ and/or TLR-IRFs in vivo. In vitro studies will employ lung epithelial cells as a primary target of in vivo exposures, permitting us t define the molecular pathways which lead to organ dysfunction. We will utilize a series of human cell-based TLR screening assays, innovative binding studies and genetically modified mice to address these aims. In addition, we will employ a custom near-infrared fluorescence (NIRF) imaging system to track SWNT in lung tissues and systemically in live animals following SWNT exposures. Results of this work will generate a comprehensive understanding of how multiple aspects of NP affect cell function and will provide reliable in vitro model systems to evaluate and engineer safe nanomaterials. PUBLIC HEALTH RELEVANCE: Nanoparticles, matter having at least one dimension less than 100 nm, are being widely used in industrial, commercial and medical applications. While inhaled ultrafine particles are already associated with pulmonary disease, limited data currently exists regarding the potential effects of nanoparticle exposure on the lung. Additional concerns surround the ability of these particles to modulate the behavior of infectious agents, such as Influenza A virus that is notorious for global pandemics. The work proposed herein will begin to elucidate how normal lung cells and tissues respond to inhaled nanoparticles and Influenza A viruses and results will expose potential long term health consequences that are crucial to the development of improved safety practices and therapeutic regimes.

描述（由适用提供）：尽管使用了广泛使用，但我们对与纳米颗粒（NP）暴露相关的生物毒性的了解有限，这引起了人们对潜在不良健康影响的担忧。吸入可能是主要的暴露途径，并且在呼吸特定物质与肺部疾病之间的关联强调了理解NP如何影响肺的关键需求。我们对单壁碳纳米管（SWNT）特别感兴趣，因为它们被广泛使用，并且可能与长期健康后果有关。围绕NP调节感染剂行为的能力的其他问题。由于NP暴露，人们对感染的敏感性增加可能会产生巨大的后果，尤其是对造成全球大流行病的臭名昭著的病毒（例如影响力）。由于现实的暴露情况可能涉及多种代理，因此触发配置的信号机制可能会导致有害的有害反应增强，从而导致更严重的健康结果。基于这些说明，该应用的总体目标是表征控制肺上皮细胞的主要免疫反应的机制，该机制单独且结合了SWNT和IAV。我们的机械重点将集中在类似收费的受体（TLR）上，因为它们是对进入体内的外国特工的早期防御线。我们将检验以下假设：不同的手性/直径的SWNT刺激TLR，从而通过激活NF-K nf-K an和irfs的转录因子而产生促炎性细胞因子。此外，SWNT和IAV的组合暴露将协同激活TLR驱动的途径，从而导致炎症和损伤增强。该前提将在3个全面的特定目的中进行测试，以（1）通过具有不同的手性包裹角和直径的SWNT以及通过IAV来确定刺激明显的TLR，以及通过IAV进行评估，（2）评估直接相互作用，（2）SWNT和IAV的IAV的相互作用，并调查了这些响应细胞在lung epepther中调节lung epepersial and lung epectional and lung epection lugn和（3），（3 SWNT和IAV的顺序暴露是由TLR-NF-KTH和/或TLR-IRF在体内介导的。体外研究将利用肺上皮细胞作为体内暴露的主要靶标，使我们定义了导致器官功能障碍的分子途径。我们将利用一系列基于人类细胞的TLR筛选测定，创新的结合研究和转基因的小鼠来解决这些目标。此外，我们将采用自定义的近红外荧光（NIRF）成像系统来跟踪肺组织中的SWNT，并在SWNT暴露后全身在活动物中跟踪。这项工作的结果将对NP的多个方面如何影响细胞功能产生全面的了解，并将提供可靠的体外模型系统来评估和设计安全的纳米材料。 公共卫生相关性：纳米颗粒，至少一个小于100 nm的物质已被广泛用于工业，商业和医疗应用中。虽然涉及的超铁颗粒已经与肺部疾病有关，但目前关于纳米颗粒暴露对肺部的潜在影响的数据有限。围绕这些颗粒调节传染剂行为的能力的其他问题，例如众所周知的全球大流行病的流感病毒。本文提出的工作将开始阐明正常的肺细胞和组织对继承纳米颗粒和流感病毒的反应，结果将使潜在的长期健康后果暴露对改善安全实践和治疗方案的发展至关重要。