Aspects of airway epithelia cell physiology

气道上皮细胞由于吸入充满污染物和微生物的大气而持续暴露于显著的外部压力下。这些上皮细胞不仅仅是一个物理屏障，它们对这些损伤做出动态反应，并具有大量的机制来保护免受吸入外来物质的损害。我的研究项目调查了气道上皮细胞能够自我防御的多种方式的各个方面。最近，作为宿主防御研究计划的一部分，我们已经开始研究两种以前未被认识的蛋白质，它们在这一领域起着关键作用，并发现了这些蛋白质在气道生物学中的新颖和令人兴奋的作用。本提案分为两个总体目标，重点关注这两种蛋白质，以便为我们理解这些因素如何影响和影响气道的宿主防御做出重大贡献。具体而言，本提案的短期目标是研究与牛皮癣蛋白和IL-22在模型气道上皮细胞中的生物学和生理学作用有关的大量不同问题。这项提案中概述的工作符合我的长期全球研究计划，该计划旨在了解这些上皮细胞如何能够很好地保护自己免受外来物质的侵害。此外，它补充了我研究的其他临床方面，为理解上皮防御的这些问题提供了基本的生物学基础，目标是将这种理解整合到创新技术和治疗方法中。本提案将使用多种不同的方法和模型细胞系，目前在我的实验室中可用，以调查有关这些蛋白质的生物学作用的一系列广泛的问题。使用细胞培养模型对这些研究有许多好处；来自气道不同部位的细胞可用于模拟各种生理相关情况；它们相对便宜且易于维护；它们可以通过基因操纵来明确地研究一种特定蛋白质的作用。所采用的方法包括分子生物学和蛋白质检测技术、伤口愈合和增殖试验、细胞活力和凋亡测量、氧化应激测量和上皮屏障功能测量。基于我们在气道上皮宿主防御领域的知识和先前的发现，我们将研究关于这两种蛋白质的一些基本问题，以及它们在保护肺部免受伤害方面的确切作用。这项工作将增加我们对该系统生物学的基础知识，为这些药物在基本细胞过程中赋予新的生物学作用，并进一步加深我们在这一领域的知识。在所有情况下，所列出的研究问题仅仅是我们调查的初始起点，并将提供重要的初步发现，以便在探索基金的整个过程中进行更全面的调查。

Airway epithelial cells are continuously exposed to significant external stresses as a consequence of inhaling atmospheric air, which is full of pollutants and micro-organisms. More than just a physical barrier, these epithelial cells dynamically respond to these insults and have a large number of mechanisms to protect against damage from inhaled foreign material. My research program investigates various aspects of the multitude of ways airway epithelial cells are able to mount to defend themselves. Recently, as part of this research program into host defence, we have begun to investigate two previously unappreciated proteins which play key roles in this area, and have discovered novel and exciting roles for these proteins in airways biology. This proposal is divided into two overall objectives focused on these two proteins, in order to contribute significantly to our understanding of how these factors affect and impact host defence of the airways. Specifically, the short term objectives of this proposal are to investigate a large number of different questions pertaining to the biological and physiological roles of psoriasin and IL-22 in model airway epithelial cells. The work outlined in this proposal fits into my long-term global research program which aims to understand how these epithelial cells are able to defend themselves so well against insult from foreign material. Furthermore, it complements other, more clinical, aspects of my research by providing a fundamental biological basis for understanding these questions of epithelial defence, with the goal of integrating this understanding into innovative technologies and therapeutic approaches. This proposal will use a variety of different methodologies and model cell lines presently available in my laboratory to investigate a wide-ranging series of questions concerning the biological role of these proteins. The use of cell culture models has a number of benefits for these studies; cells derived from different parts of the airways can be used to model a variety of physiologically relevant situations; they are relatively cheap and easy to maintain; and they can be genetically manipulated to definitively investigate the role of one particular protein in isolation. The methodologies employed will range from molecular biology and protein detection techniques, wound healing and proliferation assays, cell viability and apoptosis measurements, measurements of oxidative stress and measurements of epithelial barrier function. Building on our knowledge and previous findings in the area of airway epithelial host defence, we will investigate a number of essential questions concerning these two proteins, and what their exact roles might be in protecting the lungs against the insults it receives. This work will increase our fundamental knowledge of the biology of this system, ascribe novel biological roles for these agents in essential cellular processes, and further our knowledge in this area. In all cases, the research questions outlined are merely initial starting points for our investigations, and will provide important preliminary findings to be investigated more fully over the full course of the Discovery grant.