Targeting Airway Inflammation from Concentrated Animal Feeding Operation Dust

描述（由申请人提供）：集中动物饲养操作（cafo）的农民和工人患有与工作相关的呼吸道疾病，特别是慢性支气管炎和慢性阻塞性肺疾病（COPD）。虽然cafo中的多种物质可能导致疾病，但这些设施产生的粉尘被公认为是一种重要的呼吸道健康危害。我们以前的工作一直集中在确定CAFO粉尘导致肺部炎症的机制。重要的是，我们已经确定了这种CAFO粉尘诱导的肺部炎症机制的三个关键要素，我们提出了治疗这种重要的职业性肺部疾病的优秀治疗靶点：1)细胞因子释放，重点是tnf - α依赖的气道上皮细胞释放IL-6和IL-8，依次激活气道上皮蛋白激酶C亚型（PKC）， α随后是epsilon；2)环AMP依赖性蛋白激酶（PKA）的抗炎作用；3)促炎蛋白酶是CAFO粉尘中的触发因子。该提案概述了我们将如何使用临床前动物模型来解读靶向这三种可能抑制和/或逆转CAFO粉尘诱导的肺部疾病的机制元素的相对价值。为了达到这个目的，我们已经证明了吸入粉尘提取物会导致小鼠模型体内的呼吸道炎症，该模型具有在猪禁闭设施中工作的人所见的肺部疾病的所有突出特征。在这个更新我们提出一个策略来利用小鼠模型在临床前研究中针对上述治疗靶点的确定哪些是可行的和有效的。我们假设：CAFO粉尘诱导的肺部炎症可以通过阻断PKC异构体触发的气道细胞因子释放、激活PKA和抑制粉尘衍生的蛋白酶及其细胞靶点来治疗。我们将通过三个特定目标来验证这一假设：目标1：确定特异性靶向tnf - α， IL-6和IL-8的药物如何调节粉尘提取物诱导的体内肺部炎症。目的2：确定体外和体内增强PKA的药物，特别是治疗性β -肾上腺素能激动剂，如何抑制粉尘提取物诱导的PKC亚型激活和减轻肺部炎症。目的3：确定蛋白酶在尘埃提取物诱导的tnf - α /IL-6/IL-8体外和体内组织炎症中的重要性，并确定减轻尘埃提取物蛋白酶诱导的炎症变化的潜在靶点。我们的建议旨在提供临床前细胞、肺切片和动物数据，这些数据将促进旨在将潜在干预措施带入工作场所的转化研究。)

DESCRIPTION (provided by applicant): Farmers and workers in concentrated animal feeding operations (CAFOs) experience work-related respiratory disease, particularly chronic bronchitis and chronic obstructive pulmonary disease (COPD). Although multiple substances in CAFOs may contribute to disease, dust from these facilities is well recognized as an important respiratory health hazard. Our previous work has been focused on defining mechanisms by which CAFO dust results in lung inflammation. Importantly, we have identified three critical elements of this CAFO dust-induced lung inflammation mechanism that we propose make excellent therapeutic targets for treatment of this important occupational lung disorder: 1) cytokine release, focusing on the TNF-alpha-dependent airway epithelial cell release of IL-6 and IL-8 with sequential activation of the airway epithelial protein kinase C isoforms (PKC), alpha followed by epsilon; 2) the anti-inflammatory effects of the cyclic AMP dependent protein kinase (PKA); and 3) pro-inflammatory proteases as triggers present in CAFO dust. This proposal outlines how we will use a pre-clinical animal model to decipher the relative value of targeting these three mechanistic elements that may dampen and/or reverse CAFO dust-induced lung disease. Toward this end, we have demonstrated that inhaled dust extract causes respiratory inflammation in vivo in a mouse model that has all of the prominent features of the pulmonary disorders seen in persons working in swine confinement facilities. In this renewal we propose a strategy to utilize this mouse model in preclinical studies aimed at determining which of the therapeutic targets outlined above are feasible and efficacious. We hypothesize that: CAFO dust-induced lung inflammation is treatable by blocking PKC isoform-triggered airway cytokine release, activating PKA and inhibiting dust-derived proteases and their cellular targets. We will test this hypothesis via three specific aims: Aim 1: Establish how agents that specifically target TNF-alpha, IL-6, and IL-8 modulate dust extract-induced lung inflammation in vivo. Aim 2: Determine how agents that augment PKA, especially therapeutic beta-adrenergic agonists, dampen dust extract-induced PKC isoform activation and attenuate lung inflammation in vitro and in vivo. Aim 3: Determine the importance of proteases in dust extract-induced TNF-alpha/IL-6/IL-8 in vitro and in tissue inflammation in vivo and identify potential targets for attenuating the dust extract protease-induced inflammatory changes. Our proposal is designed to provide pre-clinical cell, lung slice, and animal data that will facilitate translational studies aimed at bringing potential interventions into the workplace. ) PUBLIC HEALTH RELEVANCE: In our previous work, we determined that dust extract from swine confined animal feeding operations causes cells lining airways to release specific inflammatory mediators, namely TNF-1, IL-6, and IL-8 via the intracellular signal protein kinase C (PKC). We have also demonstrated that this dust extract causes inflammation in a mouse model that has features similar to that seen in workers. In this application, we will perform pre-clinical studies using our mouse model to determine if targeting specific mediators (TNF-1, IL-6, and IL-8) and pathways (PKC and cAMP dependent protein kinase) as well as substances in the dust (proteases) will decrease inflammation in the lungs, with a long-term goal of developing new treatment strategies to reduce airway inflammation before it causes disease in workers.