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

在英国，呼吸道疾病的负担正在增加，自2004年以来，每五个人中就有一个死于呼吸道疾病。现在每年要花费英国国民健康保险制度66亿英镑，每年有数百万名呼吸道疾病患者需要卧床休息。因此，与我们的欧盟伙伴相比，英国的记录并不令人羡慕。呼吸道疾病并不都是由吸烟引起的，许多情况下都有遗传和环境因素(特别是空气污染)。职业和环境暴露于周围颗粒物污染会加剧既往肺部疾病患者的症状，还会降低健康人的正常肺功能。后者的结果是增加了疾病的脆弱性，降低了生活质量，损失了工作日，并增加了NHS的负担。仅在威尔士，每1000名儿童中就有57名长期因空气污染颗粒而受损，30%的哮喘急性加重与室外空气污染有关(HPA，2005)。以柴油机尾气颗粒物(DEP)为模型颗粒物(占威尔士大气污染颗粒物的80%)，研究正常患者肺组织基因表达的变化。将进行毒理基因组学实验，以确定与DEP暴露和损伤相关的分子生物标记物，并通过qPCR进行验证。通过进行差异基因表达研究，并将毒理学反应图谱与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。这些接触颗粒物的生物标志物的鉴定可能会阐明肺部疾病的机制。进行这种生物标记物研究的关键是能够使用肺模型系统，该系统允许终点数据以及急性和慢性毒物暴露窗口。贝鲁贝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是一家总部位于加的夫的中小企业，专门从事用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的微胶囊技术可以将生物化学物质和功能试剂加入到大小和剂量均一的聚合物微球中。Q Chip目前正在开发包含稳定的功能细胞的新型生物聚合物包埋剂基质，这些细胞表达与治疗相关的分子，如儿茶酚胺。通过修改微球聚合物基质以包含额外的细胞外基质蛋白，如壳聚糖、纤维连接蛋白和胶原，将产生一系列新的3D生长支撑，以稳定Berube博士的呼吸道上皮模型。这一合作项目进一步旨在研究支持在微球表面的细胞的行为、生长和寿命。为了进一步开发针对慢性呼吸道疾病的3D细胞模型，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实用性。人们认为，上述两个研究目标的成功实现将使我们能够探讨治疗(由DEP)损伤的肺组织的再生方法。成功识别遗传生物标记物和差异基因表达谱将使我们能够在体外监测和跟踪受损组织样本的疾病进展。成功开发用于呼吸系统疾病模型的生物聚合物生长支架将使我们能够研究添加生长或营养因子或种植健康、有活力的细胞的治疗性生物材料植入物。许多疾病都有可能以这种方式解决，包括帕金森氏症和脊髓损伤。微球生长载体也可能适用于其他疾病模型和涉及多潜能和多潜能细胞的再生疗法。

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.