Toxicogenomic studies of air pollutants upon a 3D supported respiratory epithelial cell model.

在英国，呼吸系统疾病的负担正在增加，自2004年以来，有五分之一的人死于呼吸系统疾病。目前，英国国家医疗服务体系每年要花费66亿英镑，每年有数百万个“卧床日”被呼吸道患者占用。因此，与我们的欧盟伙伴相比，英国的记录并不令人羡慕。呼吸系统疾病并不都是由吸烟引起的，在许多情况下，遗传和环境（特别是空气污染）因素都涉及其中。职业性和环境性接触环境颗粒污染会加重已有肺病患者的症状，也会降低健康人的正常肺功能。后者的结果是更易患病，生活质量下降，工作日减少，国民保健制度负担加重。仅在威尔士，每1000名儿童中就有57名因空气污染颗粒而长期肺功能受损，30%的哮喘急性发作与室外空气污染有关（卫生和环境保护局，2005年）。本项目以柴油尾气颗粒（DEP）为模型颗粒空气污染物（因为它占威尔士空气污染颗粒的80%），目的是研究正常患者肺组织中基因表达的变化。设计用于鉴定与深度暴露和损伤相关的分子生物标志物的毒物基因组学实验将通过qPCR进行并验证。通过进行差异基因表达研究，并将毒理学反应谱与那些特征明确的毒素进行比较，我们将了解DEP毒性的细胞机制效应。这些颗粒暴露的生物标志物的鉴定可能阐明肺部疾病的机制途径。进行此类生物标志物研究的关键是能够使用允许终点数据和急性和慢性毒物暴露窗口的肺模型系统。BeruBe博士领导卡迪夫大学的肺和粒子研究小组（LPRG），并开发了这样一个模型系统。Q Chip是一家总部位于卡迪夫的中小企业，专注于生命科学的微流体技术，特别是生物化学和生物聚合物工程的融合应用。Q Chip的微胶囊技术能够将生化试剂和功能试剂整合到均匀大小和剂量的聚合物微球中。Q Chip目前正致力于开发新型生物聚合物封装基质，内含稳定的功能细胞，这些细胞可以表达与治疗相关的分子，如儿茶酚胺。通过修饰微球形聚合物基质，使其含有额外的细胞外基质蛋白，如壳聚糖、纤维连接蛋白和胶原蛋白，将产生一系列新的3D生长支撑，以稳定BeruBe博士的呼吸上皮模型。该合作项目进一步旨在研究微球表面支撑细胞的行为、生长和寿命。为了进一步开发3D细胞模型以解决慢性呼吸道疾病，将研究支持细胞外基质和生长因子的组合，以增加细胞模型的寿命和有用性。人们认为，成功实现上述两个研究目标将使我们能够采用再生方法治疗（DEP）损伤的肺组织。成功鉴定遗传生物标志物和差异基因表达谱将使我们能够在体外监测和跟踪受伤组织样本的疾病进展。基于生物聚合物的呼吸系统疾病模型生长支持的成功开发将使我们能够研究治疗性生物材料植入物，这些植入物被添加了生长或营养因子，或被播种了健康的、有活力的细胞。许多疾病都有可能通过这种方式得到治疗，包括帕金森氏症和脊髓损伤。微球生长支持也可能适用于涉及多能和多能细胞的其他疾病模型和再生疗法。

The burden of respiratory disease in the UK is growing, with one in five people dying from it since 2004. It now costs the NHS £6.6 billion annually, with millions of 'bed days' taken up every year by respiratory patients. Consequently, the UK has an unenviable record compared to our EU partners. Respiratory illnesses are not all caused by smoking and there are genetic and environmental (particularly air-pollution) factors involved in many conditions. Occupational and environmental exposure to ambient particulate pollution exacerbates symptoms in persons with pre-existing lung disease and also reduces normal lung function in healthy individuals. The outcome of the latter is increased vulnerability to disease, reduction in quality of life, loss of work days and increased burden to the NHS. In Wales alone, 57 in 1000 children have long term impaired lung function due to air pollution particles and 30% of acute exacerbations of asthma are related to outdoor air pollution (HPA, 2005). Using diesel exhaust particles (DEP) as a model particulate air pollutant, (since it accounts for 80% of particles found in Welsh air pollution), an objective of this project is to investigate the gene-expression changes in lung tissue of normal patients. Toxicogenomic experiments designed to identify molecular biomarkers relating to DEP-exposure & injury will be performed and verified by qPCR. By performing differential gene expression studies and comparing the toxicological response profiles with those of well-characterised toxins, we will understand the cellular-mechanistic effects of DEP toxicity. The identification of such biomarkers for particle exposure may elucidate mechanistic pathways of lung disease. The key to conducting such biomarker research is the ability to use lung model systems which permit end-point data and both acute and chronic toxicant exposure windows. Dr. BeruBe leads Cardiff University's Lung and Particle Research Group (LPRG), and has developed such a model system. Q Chip is a Cardiff-based SME specialising in microfluidics technologies for life sciences, particularly convergent applications for biochemistry and biopolymer-engineering. Q Chip's micro-encapsulation technology enables the incorporation of biochemicals and functional reagents into uniformly sized & dosed polymer microspheres. Q Chip is currently working towards novel biopolymer encapsulant matrices containing stabilised, functional cells which express therapeutically-relevant molecules such as catecholamines. By modifying microspherical polymer matrices to contain additional extra-cellular matrix proteins such as chitosan, fibronectin & collagen, a new range of 3D growth supports will be generated to stabilise Dr. BeruBe's respiratory epithelium model. This collaborative project further aims to investigate the behaviour, growth and longevity of cells supported on microsphere surfaces. In order to further develop the 3D cell model to address chronic respiratory disease, combinations of supporting extracellular matrices and growth factors will be examined to increase the longevity and usefulness of the cell model. It is perceived that the successful attainment of the two research goals described above will enable us to approach regenerative methods for treatment of injured (by DEP) lung tissues. Successful identification of genetic biomarkers and differential gene-expression profiles will allow us to monitor and track disease progression in injured tissue samples in vitro. Successful development of biopolymer-based growth supports for the respiratory disease model will allow us to investigate therapeutic biomaterial implants which are dosed with growth or trophic factors or seeded with healthy, viable cells. Many diseases could potentially be addressed in this manner, including Parkinson's & spinal cord injury. Microsphere growth supports may also be applicable to other disease models and regenerative therapies involving pluripotent and multipotent cells.