Regulation of differentiation in esophageal epithelia

描述（由申请人提供）：这是我的研究项目资助（R 01）更新申请的重新提交，标题为“食管上皮分化的调节”（DK 069984），于2011年3月31日结束。 该计划的首要目标是通过对关键转录调节因子Kr的机制研究来了解食管上皮分层、炎症和癌变的调节|ppel样因子4（KLF 4）。 该建议的重要性在于（a）食管疾病的流行，其是美国和世界上最常见的疾病之一，和（B）KLF 4在控制许多组织和细胞类型（包括食管角质形成细胞）中的基本细胞过程中的重要和新兴作用。 PI是一名经验丰富的研究者，是Kr方面的专家|ppel样因子（KLF），疾病的动物模型和食管鳞状细胞生物学，并得到了一流的研究团队的支持，并得到了专家合作者的补充。 在这项提案中，我们将利用新的小鼠模型，我们已经产生，并在体外系统利用二维和三维器官型培养互补，以测试假设KLF 4是一个关键的调节成人食管鳞状上皮细胞，控制过程，如角质形成细胞迁移和分化，炎症反应，鳞状细胞癌的发展。 为了探索KLF 4对食管上皮分层、炎症和癌变的调节，我们将进行以下相关的具体目标：（1）我们将探索KLF 4在食管上皮迁移和分层中的功能，（a）通过KLF 4和KLF 4靶点WNT 5A（一种非典型Wnt）的机制、上位性研究，以及（B）通过鉴定额外的，通过功能基因组学分析KLF 4相关靶基因~（2）我们将通过（a）检测KLF 4和p53在体外和体内食管上皮细胞中的功能相互作用，和（B）确定KLF 4在食管异型增生和鳞状细胞癌中丢失的机制和（3）我们将确定KLF 4促进炎症和非炎症的机制。通过（a）评估Rho Stases在KLF 4激活NF：B中的作用，（B）检查NF：B和TNF 1抑制对来自ED-L2/KLF 4小鼠的角质形成细胞增殖的影响，和（c）将ED-L2/KLF 4小鼠与角质形成细胞特异性Ikk 2缺失小鼠杂交。 这些互补的方法将证实和扩展我们当前资助周期的发现，如初步数据和我们最近在胃肠病学上的两篇出版物所述。 此外，拟议的研究将得到一流的学术环境和PI及其团队可用的特殊资源和设施的支持。 我们预计，这些研究将提供深入了解的因素，维持正常的食管上皮稳态和被破坏的途径，在食管疾病，良性和恶性。

DESCRIPTION (provided by applicant): This is a resubmission of the renewal application for my Research Project Grant (R01) entitled "Regulation of Differentiation in Esophageal Epithelia" (DK069984), which concluded on March 31, 2011. The overarching goal of this proposal is to understand the regulation of esophageal epithelial stratification, inflammation, and carcinogenesis through mechanistic studies of the key transcriptional regulator Kr|ppel-like factor 4 (KLF4). The significance of this proposal lies in (a) the prevalence of esophageal disorders, which are among the most common ailments in the United States and the world and (b) the important and emerging roles of KLF4 in the control of essential cellular processes in numerous tissues and cell types, including esophageal keratinocyctes. The PI is an experienced investigator who is an expert in the Kr|ppel-like factors (KLFs), animal models of disease, and esophageal squamous cell biology, and is supported by a superb research team, complemented by expert collaborators. In this proposal, we will take advantage of novel mouse models, which we have generated, and complementary in vitro systems utilizing two-dimensional and three-dimensional organotypic culture to test the hypothesis that KLF4 is a critical regulator within adult esophageal squamous epithelia, controlling processes such as keratinocyte migration and differentiation, the inflammatory response, and squamous cell cancer development. To explore the regulation of esophageal epithelial stratification, inflammation, and carcinogenesis by KLF4, we will undertake the following interrelated Specific Aims: (1) We will explore the function of KLF4 in esophageal epithelial migration and stratification (a) through mechanistic, epistatic studies of KLF4 and the KLF4 target WNT5A, a non-canonical Wnt, and (b) by identification of additional, relevant KLF4 target genes through functional genomics analyses~ (2) We will investigate the role of KLF4 loss in esophageal carcinogenesis by (a) examining the functional interaction of KLF4 and p53 in esophageal epithelial cells in vitro and in vivo, and (b) identifying the mechanisms of KLF4 loss in esophageal dysplasia and squamous cell cancer~ and (3) We will determine the mechanisms by which KLF4 promotes inflammation and the non-cell autonomous proliferative response by (a) evaluating the role of Rho Stases in NF:B activation by KLF4, (b) examining the effects of NF:B and TNF1 inhibition on proliferation of keratinocytes from ED-L2/KLF4 mice~ and (c) crossing ED-L2/KLF4 mice with esophageal-specific Ikk2 null mice. These complementary approaches will confirm and extend the findings from our current funding cycle, as outlined in Preliminary Data and our two recent publications in Gastroenterology. Moreover, the proposed research will be supported by the superb and collegial intellectual environment and the exceptional resources and facilities available to the PI and his team. We anticipate that these studies will provide insight into the factors that maintain normal esophageal epithelial homeostasis and the pathways that are disrupted in esophageal diseases, both benign and malignant.