Urothelial cell response to bacterial infection

描述(申请人提供)：尿路上皮对细菌感染有一套有效的防御系统。这些包括表面分子形式的静态防御，抑制细菌附着和主动反应，以消除既定的细菌感染。这项研究的长期目标是阐明感染调节抗菌防御的机制。我们的假设是，在细菌对抗生素的耐药性不断增加的环境中，了解自然的防御机制应该允许它们的增强作为预防或治疗感染的一种手段，特别是在易受感染的人群中。肠球菌属在细菌性尿路感染的主要原因中排名第二。在该属中普遍存在的高耐药性使得肠球菌感染的治疗成为一个治疗挑战。我们实验室之前的研究已经建立了三维生长的尿路上皮模型，该模型非常接近于膀胱尿路上皮的形态和功能基因组学。在这个模型系统中，可以精确地控制尿路上皮细胞的环境，可以添加特定的调节蛋白，或者在现实的细胞培养环境中抑制或增强特定基因的作用。我们将提出两个广泛的问题：1.尿路上皮对细菌感染有何反应？2.改变尿路上皮中特定基因的表达能否调节对细菌感染的反应？这种方法将与这个合作研究小组的其他成员进行的独立细菌和动物模型研究密切结合起来。我们的模型允许机械研究，其中特定分子在尿路上皮对感染的反应中的作用可以被测试。我们在这一支持阶段的主要目标是在我们实验室扩展目前与癌症相关的cDNA阵列技术，以研究三维培养中人尿路上皮细胞在经历肠球菌感染时基因组范围内的表达变化。我们计划将Clontech系统用于由8000多个命名的人类基因组成的塑料上。其目的是根据基因随着时间的推移的行为将其聚集在一起，识别参与尿路上皮细胞对感染反应的关键信号和反应途径，并学习如何操纵和解释大规模基因组数据。这些数据将构成R01或计划项目的基本要素，并将使PI能够开发一条新的泌尿学研究路线。

DESCRIPTION (provided by applicant): The urothelium maintains an active set of defenses against bacterial infection. These include static defenses in the form of surface molecules that inhibit bacterial attachment and active responses to eliminate an established bacterial infection. The long-term aim of this study is to elucidate the mechanisms by which antibacterial defenses are modulated by infection. Our hypothesis is that in an environment of increasing bacterial resistance to antibiotics, understanding the natural mechanisms of defense should permit their augmentation as a means to prevent or treat infection, particularly in vulnerable populations. Enterococcus spp. rank second among the leading cause of bacterial UTIs. The high antibiotic resistance prevalent among this genus makes treatment of enterococcal infections a therapeutic challenge. Previous research in our laboratory has established models for urothelium grown in 3 dimensions that closely mimics both the morphology and functional genomics of the bladder urothelium. In this model system, the environment of the urothelial cells can be controlled precisely and specific modulatory proteins can be added, or the actions of specific genes can be inhibited or augmented in a realistic cell culture environment. We will ask two broad questions: 1. How does the urothelium respond to bacterial infection? 2. Can altering expression of specific genes in the urothelium modulate the response to bacterial infection? This approach will integrate closely with independent bacterial and animal model studies carried out by other members of this collaborative research group. Our model permits mechanistic investigations in which the role of specific molecules in the response of urothelium to infection can be tested. Our main objective for this period of support is to expand on current cancer-related cDNA array technology in our laboratory to investigate genome-wide changes in the expression of human urothelial cells in 3-dimensional culture as they experience infection with enterococcus. We plan to use the Clontech system on plastic consisting of over 8,000 named human genes. The aims are to cluster genes according to their behavior over time, identify the key signaling and response pathways that are involved in the response of urothelial cells to infection, and learn how to manipulate and interpret large-scale genomic data. These data will form an essential element in either an R01 or a program project and will permit the PI to develop a new line of urologic research.