NADPH oxidase-associated transition from Barrett's esophagus to adenocarcinoma

说明：请参阅说明。说明应用程序的广泛、长期目标和具体目标，并提及与健康相关的 项目(即与该机构使命的相关性)。简明扼要地描述实现这些目标的研究设计和方法。描述 你将用来追求这些目标的基本原理和技术。 此外，用两三句简单明了的话描述这项研究与公共卫生的相关性。如果应用程序得到资助，则此 原样的描述将成为公开信息。因此，不包括专有/机密信息。不要超过空格 但前提是。 胃食道反流病(GERD)影响超过十分之一的40岁以上的成年人和 每四个60岁以上的成年人中就有一个。大约10%的GERD患者发展为巴雷特食道(BE)，这是 与患食管腺癌的风险增加近30-125倍有关 (EA)。从BE到异型增生和EA的进展机制尚不清楚。因此，我们将研究 一些可能参与这一进程的途径。 在BE和EA中，ROS升高，可能在这一过程中起关键作用 从BE到EA。我们发现一种新的NADPH氧化酶NOX5-S亚型在EA细胞系中过表达， EA组织和高度不典型增生的BE粘膜。NOX5-S基因首次在人胎肾中发现 在其他组织中没有被描述过。在Seg1 EA细胞中，脉冲酸暴露引起 NOX5-S和增加过氧化氢的产生。NOX5-S来源的ROS促进细胞增殖和 减少SEG1细胞的凋亡率。然而，酸诱导的NOX5-S上调和 NOX5-S依赖的细胞增殖增加和凋亡减少的机制尚不完全清楚。 因此，我们将测试这一假设，即酸暴露通过激活NADPH氧化酶上调NADPH氧化酶 Barrett化生细胞中的Rho激酶和MAP激酶，导致自由基产生过剩，这在 TURN可能通过激活核因子kappaB进一步增强NADPH氧化酶的表达 (核因子-kB)。此外，ROS还可上调细胞周期蛋白D1和死亡结构域沉默蛋白(SODD)的表达。 细胞周期蛋白D1和SODD的表达上调可促进细胞增殖，减少细胞凋亡。 因此，BE患者存在的持续性酸反流可能导致高水平的ROS，细胞数量增加 增殖和凋亡减少，这可能导致DNA损伤和突变增加， 有助于从BE到异型增生和EA的进展。 为了验证这一假说，我们将：1)研究酸诱导表达的信号转导途径 在BE细胞系、Barrett‘s粘膜活检组织和EA细胞(Seg1、Flo和OE33)中NADPH氧化酶的表达。 我们将重点介绍Rho激酶、MAP激酶和核因子-kB的作用。2)研究NADPH的作用机制 氧化酶依赖增加细胞增殖和减少细胞凋亡，其作用集中在 Cyclin D1和SODD，并检测NADPH氧化酶衍生的ROS是否导致DNA损伤。3)审查 NOX5-S发挥功能所需的调控蛋白主要集中在rac1和p22Phox上。 进一步认识酸诱导NADPH上调的信号转导途径 导致细胞增殖增加和细胞凋亡减少的氧化酶可能提供一种合理的方法 预防EA的发展。此外，NOX5-S可能是一个潜在的早期检测肺癌的生物标志物。 食道发育不良。我们的模型也可能适用于其他与炎症相关的癌症。

DESCRIPTION: See instructions. State the application's broad, long-term objectives and specific aims, making reference to the health relatedness of the project (i.e., relevance to the mission of the agency). Describe concisely the research design and methods for achieving these goals. Describe the rationale and techniques you will use to pursue these goals. In addition, in two or three sentences, describe in plain, lay language the relevance of this research to public health. If the application is funded, this description, as is, will become public information. Therefore, do not include proprietary/confidential information. DO NOT EXCEED THE SPACE PROVIDED. Gastroesophageal reflux disease (GERD) affects more than one in ten adults over 40 years of age and one in four adults over 60. Approximately 10% of GERD patients develop Barrett's esophagus (BE), which is associated with nearly a 30-125-fold increased risk for the development of esophageal adenocarcinoma (EA). The mechanism of progression from BE to dysplasia and to EA is unknown. We will therefore examine some of the pathways that may be involved in this progression. Reactive oxygen species (ROS) are elevated in BE and EA, and may play a key role in the progression from BE to EA. We found that a novel isoform of NADPH oxidase NOX5-S is overexpressed in EA cell lines, EA tissues, and BE mucosa with high grade dysplasia. NOX5-S was first identified in human fetal kidney and has not been described in other tissues. In SEG1 EA cells pulsed acid exposure causes upregulation of NOX5-S and increases H2O2 production. NOX5-S-derived ROS contribute to increased cell proliferation and decreased apoptosis in SEG1 cells. However, the mechanisms of acid-induced NOX5-S upregulation and NOX5-S-dependent increase in cell proliferation and decrease in apoptosis are not fully understood. We will therefore test the hypothesis that acid exposure upregulates NADPH oxidases via activation of Rho kinases and MAP kinases in Barrett's metaplastic cells, causing overproduction of free radicals, which in turn may further enhance the expression of NADPH oxidases through activation of the nuclear factor kappaB (NF-kB). In addition, ROS may upregulate cyclin D1 and the silencer of the death domain (SODD). Upregulation of cyclin D1 and SODD will increase cell proliferation and decrease apoptosis in these cells. Thus the persistent acid reflux present in BE patients may cause high levels of ROS, increased cell proliferation and decreased apoptosis, which may lead to DNA damage and increased mutations, contributing to the progression from BE to dysplasia and to EA. To test this hypothesis we will: 1) Examine the signal transduction pathway of acid-induced expression of NADPH oxidases in a BE cell line, in Barrett's mucosal biopsies, and in EA cells (SEG1, FLO, and OE33). We will focus on the role of Rho kinase, MAP kinases and NF-kB. 2) Examine the mechanisms of NADPH oxidase-dependent increase in cell proliferation and decrease in apoptosis, concentrating on the role of cyclin D1 and SODD, and examine whether NADPH oxidase-derived ROS cause DNA damage. 3) Examine the regulatory proteins required for NOX5-S to function, focusing on Rac1 and p22phox. A better understanding of the signal transduction pathway of acid induced upregulation of NADPH oxidases, leading to increased cell proliferation and decreased apoptosis, may provide a rational approach to the prevention of development of EA. In addition, NOX5-S may be a potential biomarker for early detection of esophageal dysplasia. Our model may also be applicable to other inflammation-associated cancers.