The role of microenvironment in esophageal epithelial homeostasis

描述(由申请人提供)：本提案描述了一个两年的综合导师培训计划，以及一个三年的独立计划，以发展胃肠病学的学术基础科学研究生涯。PI已经完成了她的细胞生物学博士学位，并试图以她现有的研究经验和技能为基础，在需要额外的基本多学科培训的研究领域成为一名成功的独立调查员。PI将获得独特的技能，研究关键炎症介质IKK在食道微环境调节中的作用。虽然细胞因子、趋化因子和炎症介质的激活在食道疾病中已经被发现，但关于这些疾病中这种激活的分子机制的信息很少。为了剖析相关途径，PI将通过正规课程、相关技术技能的掌握以及这些领域专家的指导，将免疫学和肿瘤微环境的概念整合在一起。候选人的导师乔纳森·卡茨博士是基因工程小鼠疾病模型和食道鳞状细胞生物学方面的专家。候选人的共同导师Anil Rustgi博士和Sandra Ryeom博士在转录调节、信号转导、三维培养、血管生成和微环境调节方面提供了额外的专业知识。一个由NIH资助的具有广泛专业知识的主要调查人员组成的卓越咨询委员会已经成立，以提供科学和专业指导。在这里，我们将利用新的小鼠模型和互补的体外系统，利用3D培养系统来测试这一假说，即食道上皮细胞内IKK途径的激活会产生一个加强食道不典型增生、癌症和其他疾病的微环境。为了探索这些进程，我们将实现三个相互关联的具体目标。在目标1(K99期)，我们将确定上皮样IKK信号在微环境和食道上皮-内皮细胞相互作用中的作用。这将使用一种新的转基因小鼠模型和3D组织背景下的原代食道上皮细胞来进行。在目标2(K99/R00期)，我们将确定上皮IKK信号在限制食道基质肌成纤维细胞扩张中的需求。在这里，我们将利用食道特异的IKK基因敲除小鼠和3D培养。在目标3(R00期)，我们将确定STAT3激活和IKK？/NF？B信号在食道上皮细胞炎症反应中的功能相互作用。为了检验这些相互作用，我们将使用与STAT3 Flosed小鼠杂交的ikk？基因敲入小鼠。拟议的研究将得到一流的和合作的知识环境以及特殊的资源和设施，可供国际和平研究所支持。我们预计这些研究将为调节正常的食道上皮稳态的因素、微环境以及在食道疾病中被破坏的途径提供洞察力，无论是良性的还是恶性的。

DESCRIPTION (provided by applicant): This proposal describes a two year integrated mentored training program followed by a three year independent program for the development of an academic basic science research career in gastroenterology. The PI has completed her Ph.D. in Cell Biology and seeks to build on her existing research experience and skills to become a successful independent investigator in an area of research that requires additional essential multi- disciplinary training. The PI will acquire unique skills set to study the role of he key inflammatory mediator IKK¿ in the regulation of the esophageal microenvironment. While activation of cytokines, chemokines, and inflammatory mediators has been identified in esophageal diseases, little information is available about the molecular mechanisms of this activation in these diseases. To dissect the relevant pathways, the PI will integrate concepts from immunology and the tumor microenvironment through formal coursework, mastering of relevant technical skills, and mentorship by experts in these fields. The candidate's Mentor, Dr. Jonathan Katz, is an expert in genetically-engineered mouse models of disease and esophageal squamous cell biology. The candidate's Co-Mentors, Dr. Anil Rustgi and Dr. Sandra Ryeom, provide additional expertise in transcriptional regulation, signal transduction, three-dimensional culture, angiogenesis, and the regulation of the microenvironment. A superb advisory committee composed of leading NIH-funded investigators with broad expertise has been formed to provide scientific and professional guidance. Here, we will take advantage of new mouse models and complementary in vitro systems utilizing 3D culture system to test the hypothesis that activation of the IKK¿ pathway within esophageal epithelial cells produces a microenvironment that potentiates esophageal dysplasia, cancer, and other diseases. To explore these processes, we will undertake three interrelated Specific Aims. In Aim 1 (K99 phase), we will define the role of epithelial IKK¿ signaling in the microenvironment and in epithelial-endothelial cell interactions i the esophagus. This will be undertaken using a novel transgenic mouse model and primary esophageal epithelial cells in a 3D tissue context. In Aim 2 (K99/R00 phases), we will determine the requirement for epithelial IKK¿ signaling in limiting expansion of esophageal stromal myofibroblasts. Here, we will utilize esophageal-specific IKK¿ knockout mice and 3D culture. In Aim 3 (R00 phase), we will determine the functional interplay of STAT3 activation and IKK¿/NF¿B signaling in the inflammatory response of esophageal epithelial cells. To examine these interactions, we will employ IKK¿ knock-in mice that are crossed with STAT3 floxed mice. The proposed research will be supported by the superb and collegial intellectual environment as well as the exceptional resources and facilities available to the PI. We anticipate that these studies will provide insight into the factors that regulate normal esophageal epithelial homeostasis, the microenvironment, and the pathways that are disrupted in esophageal diseases, both benign and malignant.