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Computational and Cell Culture Models for Mucus Clearance

粘液清除的计算和细胞培养模型

基本信息

批准号：
7936939
负责人：
RICHARD SUPERFINE
金额：
$ 47.36万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-21 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7936939
关键词：
3-Dimensional Address Affect Air Allergens Animal Model Animals Area Asthma Automation Bacterial Infections Benchmarking Biochemical Biochemistry Biological Assay Biological Models Biophysics Breathing Cell Culture System Cell Culture Techniques Cell membrane Cell surface Cells Cereals Chemicals Chronic Chronic Obstructive Airway Disease Cilia Complement Complex Computer Simulation Computers Coughing Coupled Coupling Cystic Fibrosis Development Diffuse Disease Effectiveness Environment Environmental Monitoring Epithelial Cells Face Failure Floods Force of Gravity Generations Goals Grant Growth Health Height Hereditary Disease Human In Vitro Infection Infectious Agent Link Liquid substance Liver Lung Mathematics Measurement Membrane Microfluidics Microscope Modeling Mucociliary Clearance Mucous body substance Natural Immunity Organ Particulate Physics Positioning Attribute Race Regulation Reliance Research Research Personnel Respiratory physiology Rheology Role Screening procedure Speed Sterility Structure Structure of parenchyma of lung Surface System Technology Testing Theoretical model Thick Time Toxic Environmental Substances Toxic effect Toxicant exposure Toxicity Tests Toxin Tube Vision assault assay development base body system cell type density design effective therapy environmental stressor hazard high throughput screening human stem cells improved interfacial mathematical model novel pathogen predictive modeling prevent research study response shear stress success virtual viscoelasticity

项目摘要

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (06) Enabling Technologies and specific Challenge Topic 06-ES-102*: 3-D or virtual models to reduce use of animals in research: Creation of miniature multi-cellular organs for high throughput screening for chemical toxicity testing. Development of novel micro-scale systems of multiple cell types that replicate the macro-scale structure and function of major organ systems in response to environmental stressors linked with development of computational models of organ system function can accelerate testing of the multitude of chemicals in our environment for toxicity. Research which furthers the generation of 3-D biological models will provide new assays for rapid screening of toxicity in organs such as the lung and liver. Cell types, such as human stem cells, used in these systems would reduce the use of animals and improve our assessment of chemical hazards in the environment. Contact: Dr. David Balshaw, balshaw@niehs.nih.gov, (919) 541-2448 Title: Computational and Cell Culture Models of Mucus Clearance. Summary: The lung inhales over 1 million infectious and toxic agents each day. The defense of the lung begins with the layer of mucus that lines the epithelial cell layer. When these assaults land at the mucus/air inter- face, the race between infection and clearance begins. The particulates, pathogens and allergens are in a diffusive race for the cell membrane before they can be cleared by the entraining flow of mucus. To provide effective clearance, the body must accomplish several goals. First it must provide the barrier layer of mucus with a height and viscoelasticity that impose a significant transit time for the infectious agents. Second, propulsion mechanisms through cilia or airflow must move the mucus with a speed that clears the agents before they can diffuse to the cell membrane. Third, the viscoelasticity of the mucus must be sufficient to prevent flooding of the airway, yet be fluid enough to allow for transport by cilia and airflow. The goal of this project is to generate computational and cell culture based models for mucus clearance. Since failure of clearance can be under- stood as the first step to a cascade of lung failure scenarios, the establishment of effective model systems for toxicity testing is critical. The use of a cell culture based model allows the inclusion of complex biochemical and immunological responses. The use of ciliated cultures that have been shown to generate and maintain appropriate mucus layers, and to generate a coordinated array of cilia for propulsion, is the starting point for our model system. We will generate the first ciliated cell culture systems that place the cells into a geometry that replicates essential features of the lung: directional, linear flow with converging cross section that can be challenged for vertical transport against gravity. This model system will have the potential to act as a sensitive as- say for environmental effectors that compromise mucus height regulation, mucus rheology, cilia density and co- ordination. We will further develop the assay within a microfluidic system that has twelve isolated assays operating in parallel. This will bring this sophisticated cell based assay to medium-throughput screening. Beyond biophysical models, we require a computational model so that we may predict the consequences of environmental assaults and design effective therapies. We will develop theoretical models that in- corporate the propulsion mechanisms of cilia and of airflow. The latter is operative during effective clearance maneuvers such as cough, and it is understood that even in the absence of cilia propulsion, cough can maintain sterile airways. However, there is currently no predictive model of the role of mucus rheology and layer thickness, cilia effectiveness and airflow in producing sufficient clearance to maintain healthy lung function. By combining a team of researchers from Applied Mathematics, Physics, Biochemistry and Biophysics, and the UNC Cystic Fibrosis Center, the goal of this project is to develop cell-based biophysical assays that can test environmental assaults for their role in compromising mucus clearance, and use them to establish a computational model for clearance that will have the potential for creating in-silico testing of toxins. The goal of this targeted two year project is to jump-start technical advances, in cell cultures and mathematical modeling, that will contribute to the vision of effective, efficient and physiologically accurate toxicity assays. The availability of a cell culture-based clearance assay coupled with an in- silico computational model will reduce the reliance on animal models and more accurately predict the consequences of toxins on human health.

描述（由申请人提供）：本申请涉及广泛的挑战领域（06）使能技术和特定的挑战主题06-ES-102*：减少研究中使用动物的3-D或虚拟模型：创建用于化学毒性试验高通量筛选的微型多细胞器官。开发新型的多细胞类型的微尺度系统，复制主要器官系统的宏观尺度结构和功能，以响应与器官系统功能的计算模型的开发相关的环境应激源，可以加速对我们环境中多种化学品毒性的测试。进一步生成3D生物模型的研究将为快速筛选肺和肝等器官中的毒性提供新的测定方法。在这些系统中使用的细胞类型，如人类干细胞，将减少对动物的使用，并改善我们对环境中化学危害的评估。联系人：大卫巴尔肖博士，balshaw@niehs.nih.gov，（919）541-2448标题：粘液清除的计算和细胞培养模型。肺每天吸入超过100万种传染性和毒性物质。肺的防御始于上皮细胞层的粘液层。当这些攻击降落在粘液/空气界面时，感染和清除之间的竞赛就开始了。微粒、病原体和过敏原在被粘液的夹带流清除之前，处于对细胞膜的扩散竞争中。为了提供有效的清除，该机构必须实现几个目标。首先，它必须为粘液屏障层提供一定的高度和粘弹性，以使感染因子有显著的通过时间。第二，通过纤毛或气流的推进机制必须以一定的速度移动粘液，以便在介质扩散到细胞膜之前清除介质。第三，粘液的粘弹性必须足以防止气道的泛滥，但仍具有足够的流动性以允许通过纤毛和气流运输。该项目的目标是生成基于计算和细胞培养的粘液清除模型。由于清除失败可以被理解为肺衰竭级联的第一步，因此建立有效的毒性试验模型系统至关重要。使用基于细胞培养的模型允许包括复杂的生物化学和免疫学应答。使用纤毛文化，已被证明可以产生和维持适当的粘液层，并产生一个协调的纤毛阵列的推进，是我们的模型系统的起点。我们将产生第一个纤毛细胞培养系统，将细胞置于复制肺基本特征的几何形状中：具有会聚横截面的定向线性流动，可以挑战垂直运输对抗重力。该模型系统将有可能作为一个敏感的假设，环境效应，损害粘液高度调节，粘液流变学，纤毛密度和协调。我们将进一步开发一个微流控系统内的分析，有12个独立的分析并行操作。这将使这种复杂的基于细胞的测定达到中等通量筛选。除了生物物理模型之外，我们还需要一个计算模型，以便我们可以预测环境攻击的后果并设计有效的治疗方法。我们将发展理论模型，包括纤毛和气流的推进机制。后者在有效的清除操作（例如咳嗽）期间起作用，并且应当理解，即使在没有纤毛推进的情况下，咳嗽也可以保持无菌气道。然而，目前还没有粘液流变学和层厚度、纤毛有效性和气流在产生足够的清除率以维持健康的肺功能中的作用的预测模型。通过结合来自应用数学，物理学，生物化学和生物物理学的研究人员团队，以及Cystic Fibrosis Center，该项目的目标是开发基于细胞的生物物理测定，可以测试环境攻击在损害粘液清除中的作用，并使用它们建立清除的计算模型，这将有可能创建毒素的计算机测试。这个为期两年的目标项目的目标是启动细胞培养和数学建模方面的技术进步，这将有助于实现有效，高效和生理准确的毒性测定。基于细胞培养的清除试验结合计算机模拟计算模型的可用性将减少对动物模型的依赖，并更准确地预测毒素对人类健康的影响。