GENETIC AND CELLULAR BASIS OF H. PYLORI PATHOGENESIS

描述(改编自申请人的摘要)：建议的目标 研究目的是更好地了解幽门螺杆菌与其相互作用 人类的主人。幽门螺杆菌是胃炎和溃疡病的病原体， 而感染是胃病发生的重要危险因素。 癌症。在之前的研究中，这些研究人员已经表明，幽门螺杆菌CagA 被转运到宿主细胞，然后在酪氨酸上被磷酸化 残留物由宿主细胞的激酶活性决定。CagA插入宿主细胞的研究 而其随后的磷酸化与重组有关 宿主细胞的细胞骨架，以及宿主细胞形态的戏剧性变化。 研究人员建议检查CagA插入宿主细胞和 插入CagA后的一系列事件。此外，他们还提议 通过突变改变CagA的结构以确定特异性的作用 引起细胞变化的蛋白质结构域。 在第二阶段的工作中，这些调查人员已经获得了 幽门螺杆菌基因组和宿主基因阵列的人和小鼠的基因。他们 建议用幽门螺杆菌DNA芯片检测临床菌株的基因分型 从明确的流行病学研究中分离出来的菌株，以确定 特定的基因或基因组与离散的临床相关 像溃疡疾病或恶性肿瘤这样的症状。建议的主旨是 工作将集中在使用DNA阵列来跟踪两者的基因转录 细菌和宿主基因在人类极化培养物感染中的作用 上皮细胞。他们假设这将允许识别出 新的毒力基因类别。所测量的主机响应的顺序 建议通过基因转录来更好地了解寄主 细菌在感染过程中利用的细胞途径。最后，一个 将使用称为微阵列转座子标记幽门螺杆菌(Matt)的方法，该方法 允许从选择性突变中识别特定类别的突变 环境，包括受感染的细胞培养物和动物。

DESCRIPTION (Adapted from the Applicant's Abstract): The goal of the proposed research is to understand better the interaction between H. pylori and its human host. H. pylori is the causative agent of gastritis and ulcer disease, and infection is an important risk factor for the development of gastric cancer. In previous studies, these investigators have shown that H. pylori CagA is transported into the host cell, and is then phosphorylated on tyrosine residues by host cell kinase activity. The insertion of CagA into host cells and its subsequent phosphorylation is associated with reorganization of the host cell cytoskeleton, as well as a dramatic change in host cell morphology. The investigators propose to examine the insertion of CagA into host cells and the sequence of events following CagA insertion. In addition, they propose to alter the structure of CagA by mutagenesis to determine the role of specific protein domains in bringing about cellular changes. In a second phase of work, these investigators have obtained DNA arrays of the H. pylori genome and host gene arrays of both human and mouse genes. They propose to use the H. pylori DNA array to examine the genotype of clinical isolates from well-defined epidemiological studies to determine whether particular genes or groups of genes are associated with discrete clinical syndromes like ulcer disease or malignancy. The major thrust of the proposed work will focus on the use of DNA arrays to follow gene transcription of both bacterial and host genes during infection of polarized cultures of human epithelial cells. They hypothesize that this will permit the identification of new classes of virulence genes. The sequence of the host response as measured by gene transcription is suggested to permit better understanding of the host cell pathways that are exploited by the bacteria during infection. Finally, a method called microarray transposon tagged H. pylori (MATT) will be used, which permits the identification of specific classes of mutation from selective environments, including infected cell cultures and animals.