Molecular Mechanisms Regulating Intestinal Homeostasis

描述（由申请方提供）：肠粘膜通过时空调节沿着肠隐窝-管腔轴的增殖、分化、迁移和细胞死亡来保持其完整性。协调这些事件的分子机制，特别是在损伤修复的背景下，还没有得到很好的理解。此外，目前尚不清楚 这些过程的扰动如何影响细胞结的形成和屏障功能。Kruppel样因子5（KLF 5）是转录因子家族中的一员，其功能在于调节细胞内的转录水平。 调节多种细胞过程，包括发育、增殖和分化。 KLF 5在肠上皮的增殖组分中高度表达，并且已经显示在体外和体内促进肠上皮细胞的增殖。最近的数据表明，KLF 5是维持肠隐窝结构和上皮屏障完整性所必需的。此外，最近显示KLF 5对于结肠炎的葡聚糖硫酸钠（DSS）小鼠模型中的粘膜愈合是必需的。基于这些结果，我们假设KLF 5控制上皮细胞沿着腔轴的更新和成熟，并且是肠上皮完整性所必需的。为了验证这一假设，我们提出了以下目标：（I）确定KLF 5在调节肠腺腔轴上皮细胞增殖、迁移和成熟中的作用。遗传小鼠模型和细胞培养系统将用于异位表达或抑制KLF 5，以探索KLF 5在调节肠上皮稳态过程中的作用。(II)确定KLF 5在维持肠功能中的需要量 上皮屏障功能将确定KLF 5表达对肠通透性和顶端连接复合物形成的影响。(III)研究KLF 5在肠上皮损伤修复中的作用。这些实验将测试KLF 5水平的增加是否可以改善DSS诱导的结肠炎的结果和上皮愈合。将测试几种增加小鼠结肠中KLF 5表达的方法，包括腺病毒递送、纳米颗粒递送和转基因表达。还将进行使用具有可诱导的丝氨酸特异性Klf 5缺失的小鼠的相互研究，以测试KLF 5是否是肠损伤修复所必需的。这些研究的长期目标是阐明在组织更新和损伤修复的情况下调节肠上皮完整性的分子机制。拟议的研究将有助于更好地了解协调增殖，迁移和分化过程以维持肠道结构和屏障功能的因素。特别是，这些研究将探索KLF 5在上皮成熟、迁移和细胞连接形成过程中的新作用，以及KLF 5作为炎症性肠道疾病等疾病治疗靶点的可能性。

DESCRIPTION (provided by applicant): The intestinal mucosa maintains its integrity through spatiotemporal regulation of proliferation, differentiation, migration, and cell death along the crypt-luminal axis of the intestine. The molecular mechanisms that coordinate these events, particularly in the context of injury repair, are not well understood. Furthermore, it is not known how perturbations of these processes impact cell junction formation and barrier function. Kruppel- like factor 5 (KLF5) is a member of a family of transcription factors that function in the regulation of diverse cellular processes including development, proliferation, and differentiation. KLF5 is highly expressed in the proliferative component of the intestinal epithelium, and has been shown to promote proliferation of intestinal epithelial cells in vitro and in vivo. Recent dat from mice with intestine-specific deletion of Klf5 indicate that KLF5 is required for maintenance of intestinal crypt architecture and epithelial barrier integrity. In addition, it was shown recenty that KLF5 is necessary for mucosal healing in the dextran sulfate sodium (DSS) mouse model of colitis. Based on these results, we hypothesize that KLF5 controls renewal and maturation of epithelial cells along the crypt-luminal axis and is required for intestinal epithelial integrity. he following aims have been developed to test this hypothesis: (I) Define the role of KLF5 in regulating proliferation, migration, and maturation of epithelial cells along the crypt-luminal axi of the intestine. Genetic mouse models and cell culture systems will be used for ectopic expression or inhibition of KLF5 to explore the role of KLF5 in regulating processes involved in intestinal epithelial homeostasis. (II) Determine the requirement of KLF5 in maintaining intestinal epithelial barrier function. The effects of KLF5 expression on intestinal permeability and the formation of apical junctional complexes will be determined. (III) Examine the role of KLF5 in intestinal epithelial wound repair. These experiments will test whether increased levels of KLF5 can improve outcome and epithelial healing in DSS-induced colitis. Several methods to increase KLF5 expression in the mouse colon will be tested, including adenoviral delivery, nanoparticle delivery and transgenic expression. Reciprocal studies using mice with inducible intestine-specific deletion of Klf5 will also be conducted to test whether KLF5 is necessary for repair of intestinal injury. The long-term goal of these studies is to elucidate the molecular mechanisms that regulate intestinal epithelial integrity, both in situations of tissue renewal and injury repar. The proposed studies will contribute to a better understanding of factors that coordinate the processes of proliferation, migration, and differentiation to maintain intestinal architecture and barrier function. In particular, these studies will explore novel roles for KLF5 in the processes o epithelial maturation, migration, and cell junction formation, and the possibility for KLF5 as a therapeutic target in disease conditions such as inflammatory bowel disease.