Chronic Granulomatous Lung Inflammation Elicited by Carbon Nanotubes

碳纳米管引起的慢性肉芽肿性肺部炎症

• 批准号: 8433120
• 负责人: Mary Jane Thomassen
• 金额: $ 36.82万
• 依托单位: EAST CAROLINA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8433120
• 关键词: Acute; Address; Adverse effects; Agonist; Air; Alveolar Macrophages; Animal Model; Animals; Antigens; Appearance; Area; Bicycling; Biological Assay; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Burn injury; Carbon; Carbon Nanotubes; Chronic; Cytokine Receptors; Data; Depressed mood; Diagnosis; Diesel Fuels; Disease; Environment; Environmental Health; Environmental Impact; Environmental Risk Factor; Etiology; Event; Experimental Animal Model; Experimental Models; Exposure to; Gene Deletion; Genes; Goals; Granuloma; Granulomatous; Human; Image; Individual; Industry; Inflammation; Inflammatory; Investigation; Knockout Mice; Learning; Ligands; Link; Lung; Lung Inflammation; Lung diseases; Marketing; Methane; Methodology; Modeling; Monitor; Mus; Nanotechnology; Nanotubes; Natural Gas; Occupational Health; PPAR gamma; Pathway interactions; Patients; Peroxisome Proliferator-Activated Receptors; Physicians; Plasmids; Propane; Publishing; Pulmonary Sarcoidosis; Quantitative Reverse Transcriptase PCR; Reagent; Reporting; Role; Sarcoidosis; Sepharose; Stimulus; Structure of parenchyma of lung; Students; Subfamily lentivirinae; Sunscreening Agents; Therapeutic Effect; Toxic effect; Transduction Gene; Up-Regulation; Wild Type Mouse; Wood material; World Trade Center disaster; base; chemokine; chemokine receptor; consumer product; experience; glucose metabolism; human disease; injured; laser capture microdissection; lipid metabolism; macrophage; nanomaterials; nanoparticle; novel; pathogen; public health relevance; pulmonary granuloma; receptor; response; rosiglitazone; vapor

DESCRIPTION (provided by applicant): Use of nanomaterials in manufactured consumer products is a rapidly expanding area but potential toxicities have not been established. Combustion-generated multiwall carbon nanotubes (MWCNT) or nanoparticles are ubiquitous in non-manufacturing environments and detectable in vapors from diesel fuel, methane, propane and natural gas. Carbon nanotubes induce granulomas or inflammation in some experimental animals. Pulmonary granulomas in human disease may form in response to environmental stimuli such as intracellular pathogens, inert materials, and organic antigens. In sarcoidosis, a prototypical granulomatous disease, etiology remains obscure. Multiple environmental risk factors have been linked to sarcoidosis, including exposure to wood-burning stoves, fireplaces, and firefighting - conditions that might favor carbon nanotube formation in ambient air. Investigation of putative nanotube environmental factors linked to sarcoidosis has not been done using animal models. This proposal will utilize a novel murine MWCNT-elicited, chronic granuloma model to investigate the impact of carbon nanotubes on specific host receptors, peroxisome proliferator-activated receptor? (PPAR?) and chemotactic cytokine receptor 5 (CCR5), reported to be dysregulated in sarcoidosis lung. PPAR?, a negative regulator of inflammation, is constitutively expressed in healthy alveolar macrophages but deficient in severe sarcoidosis. In contrast, proinflammatory CCR5 ligands are not found in healthy lung but are elevated in sarcoidosis. We noted similar findings in lungs of MWCNT-instilled mice: PPAR? is depressed and CCR5 ligands are elevated. Based on these data, we hypothesize that MWCNT repress PPAR? and upregulate CCR5 pathways to form pulmonary granulomas with chronic inflammation. Specific Aim 1 will determine the role of PPAR? in the MWCNT model by monitoring granuloma size and numbers in: (a) macrophage-specific PPAR?-null versus wild-type mice; and (b) mice treated with lentivirus-PPAR? plasmids, PPAR? agonist rosiglitazone; or PPAR? antagonist BADGE versus untreated mice. Specific Aim 2 will examine CCR5 involvement in MWCNT granulomas by quantifying CCR5 receptor and chemokines in bronchoalveolar lavage (BAL) derived alveolar macrophages, BAL fluids, and in granulomatous foci isolated by laser-capture microdissection [LCM], from untreated wild-type and PPAR? null mice versus mice treated with a CCR5 blocker. This study will afford students opportunities to gain experience with animal models of lung disease, learn effects of gene deletion in disease, and interact with physicians to learn environmental causes of human lung disease and how human lung disease is diagnosed and treated. Students will also learn lentivirus plasmid construction for gene transduction, quantitative RTPCR, LCM, Luminex assays, and imaging methodology. In summary, this unique investigation of MWCNT-elicited granulomatous lung disease is well suited for student participation in an area with environmental impact on human disease.

描述（由申请人提供）：纳米材料在消费产品中的使用正在迅速扩大，但潜在的毒性尚未确定。燃烧产生的多壁碳纳米管 (MWCNT) 或纳米颗粒在非制造环境中无处不在，并且可以在柴油、甲烷、丙烷和天然气的蒸气中检测到。碳纳米管会在一些实验动物中诱发肉芽肿或炎症。人类疾病中的肺肉芽肿可能是对环境刺激（如细胞内病原体、惰性物质和有机抗原）的反应而形成的。结节病是一种典型的肉芽肿性疾病，其病因仍不清楚。多种环境风险因素与结节病有关，包括接触燃木炉、壁炉和消防——可能有利于环境空气中碳纳米管形成的条件。尚未使用动物模型对与结节病相关的假定纳米管环境因素进行调查。该提案将利用新型小鼠多壁碳纳米管引发的慢性肉芽肿模型来研究碳纳米管对特定宿主受体（过氧化物酶体增殖物激活受体）的影响？ (PPAR?) 和趋化细胞因子受体 5 (CCR5)，据报道在肺结节病中失调。 PPAR？是炎症的负调节因子，在健康的肺泡巨噬细胞中持续表达，但在严重的结节病中缺乏。相比之下，促炎性 CCR5 配体在健康肺部中未发现，但在结节病中升高。我们在 MWCNT 注入小鼠的肺部发现了类似的发现：PPAR？被抑制并且CCR5配体被升高。根据这些数据，我们假设 MWCNT 会抑制 PPAR？并上调 CCR5 通路，形成伴有慢性炎症的肺肉芽肿。具体目标1将决定PPAR的作用？在 MWCNT 模型中，通过监测以下动物的肉芽肿大小和数量：(a) 巨噬细胞特异性 PPAR?-null 与野生型小鼠； (b) 用慢病毒-PPAR 治疗的小鼠？质粒、PPAR？激动剂罗格列酮；还是PPAR？拮抗剂 BADGE 与未治疗的小鼠的比较。具体目标 2 将通过量化支气管肺泡灌洗 (BAL) 衍生的肺泡巨噬细胞、BAL 液中的 CCR5 受体和趋化因子，以及通过激光捕获显微切割 [LCM] 从未经处理的野生型和 PPAR 分离的肉芽肿病灶中检查 CCR5 与 MWCNT 肉芽肿的关系？无效小鼠与用 CCR5 阻断剂治疗的小鼠。这项研究将为学生提供获得肺部疾病动物模型经验的机会，了解基因缺失对疾病的影响，并与医生互动以了解人类肺部疾病的环境原因以及如何诊断和治疗人类肺部疾病。学生还将学习用于基因转导的慢病毒质粒构建、定量 RTPCR、LCM、Luminex 测定和成像方法。总之，这项对多壁碳纳米管引起的肉芽肿性肺病的独特研究非常适合学生参与环境对人类疾病影响的领域。

项目成果

The role of PPARγ in carbon nanotube-elicited granulomatous lung inflammation.

PPARγ 在碳纳米管引起的肉芽肿性肺部炎症中的作用。

• 发表时间: 2013-01-23
• 影响因子: 5.8
• 作者: Huizar, Isham;Malur, Anagha;Patel, Janki;McPeek, Matthew;Dobbs, Larry;Wingard, Christopher;Barna, Barbara P;Thomassen, Mary Jane
• 通讯作者: Thomassen, Mary Jane