Mechanisms of Cellular Adaptation in Cystitis

 描述(由申请人提供)：以下建议概述了我的计划，成为一名独立研究人员，专注于为膀胱炎患者寻找解决方案。我是范德比尔特大学泌尿外科的助理教授，接受过儿科泌尿外科的奖学金培训。我将在接下来的5年职业生涯中致力于发展成为一名独立科学家的科学和专业能力的初步目标。膀胱炎，或膀胱炎，是一种流行的问题，每年由于尿路感染、膀胱梗阻、放射和化疗而影响数百万美国人。膀胱炎导致膀胱损伤的患者会出现大小便失禁、膀胱疼痛、血尿甚至肾脏损伤。许多膀胱炎患者最终将需要侵入性膀胱手术。在了解膀胱内的尿路上皮细胞如何在损伤后适应并在随后的炎症中做出反应方面，存在着一个基本的知识缺口。对这个问题有更深入的了解，对于开发减轻膀胱炎造成的短期和长期损害的治疗方法非常重要。我作为一名独立研究人员的发展将使我能够提出重要的科学问题，然后寻求答案，以改善膀胱炎患者的生活。我将努力通过一个有指导的科学实验和职业发展活动的结构化计划来建立我的独立能力。这些活动将包括定期的科学互动、研究生院的课程工作、技术培训、智力互动、校园研讨会、补助金撰写课程和实验室会议。导师将是这个项目的核心，一个由经验丰富的科学家组成的咨询委员会将指导我的科学和职业发展。我的机构有大量的资源来促进职业发展，我将充分利用这些机会。新的证据表明，缺氧诱导因子(HIF)途径的急性激活在损伤和炎症的背景下对上皮细胞有利。我们已经产生了初步数据，表明HIF激活发生在损伤后的膀胱中。解锁HIF信号通路潜在益处的关键在于含有Pro-4-羟基酶结构域(PHD)的蛋白质。PHD是上游的氧敏感酶，能拮抗HIF活性。来自多种不同器官损伤模型的新证据表明，抑制PHD活性会导致HIF途径的激活，从而促进上皮细胞的存活和保护。我们已经证明，抑制PHD在尿路上皮损伤和膀胱炎的环境中具有保护作用。基于我们的初步数据和已发表的其他器官的数据，我们假设损伤后的尿路上皮适应依赖于HIF介导的基因转录，而PHD抑制是这一过程的关键第一步。我们将通过执行三个具体目标来检验中心假设。目的1利用小鼠膀胱炎模型，研究抑制单个PHD2家族成员如何保护膀胱免受损伤。目的2探讨低氧诱导因子(HIF)介导的尿路上皮细胞在PHD抑制条件下的转录反应。最后，Aim 3将评估尿路上皮细胞特异性HIF表达在损伤性膀胱炎中的整体作用。

 DESCRIPTION (provided by applicant): The following proposal outlines my plan to become an independent researcher focused on finding solutions for patients with bladder inflammation. I am an Assistant Professor in the Department of Urologic Surgery at Vanderbilt University with fellowship training in Pediatric Urology. I will devote the next 5 years of my career to the primar goal of developing the scientific and professional capacity to be an independent scientist. Cystitis, or bladder inflammation, is a prevalent problem that affects millions of Americans every year as a result of urinary tract infections, bladder obstruction, radiation, and chemotherapy. Patients who develop bladder damage from cystitis suffer from incontinence, bladder pain, hematuria and even renal damage. Many patients with bladder inflammation will ultimately require invasive bladder surgery. A fundamental knowledge gap exists in understanding how the urothelial cells that line the bladder adapt following injury and respond during the ensuing inflammation. Gaining increased insight into this problem is important for developing treatments to mitigate the short and long term damage caused by cystitis. My development into an independent researcher will enable me to ask the important scientific questions and then seek out the answers to improve the lives of patients with bladder inflammation. I will work to establish my independent capacity through a structured program of mentored scientific experiments and career development activities. These activities will consist of regular scientific interaction, graduate school coursework, technical training, intellectual interactions, on-campus seminars, grant writing courses and laboratory meetings. Mentorship will be the centerpiece of this project and an advisory committee of experienced scientists will guide my scientific and career development. My institution has a multitude of resources to foster career development and I will make full use of those opportunities. Emerging evidence indicates that acute activation of the Hypoxia inducible factor (HIF) pathway is beneficial for epithelial cells in the setting of injury and inflammation. We have generated preliminary data indicating that HIF activation occurs in the bladder following injury. The key to unlocking the potential benefits of the HIF signaling pathway lies in Prolyl-4-hydroxylase domain-containing (PHD) proteins. PHDs are the up-stream oxygen sensitive enzymes that antagonize HIF activity. New evidence from multiple different models of organ injury shows that inhibiting PHD activity leads to activation of the HIF pathway and subsequently promotes epithelial survival and protection. We have shown that PHD inhibition is protective in the setting of urothelial injury and cystitis. Based on our preliminary data and published data in other organs, we hypothesize that urothelial adaptation after injury is dependent upon HIF-mediated gene transcription and that PHD inhibition is the critical first step in this process. We will test the central hypothesis by executing three specifi aims. Aim 1 seeks to understand how inhibition of individual PHD2 family member protects the bladder from injury using mouse models of cystitis. Aim 2 will explore the HIF-mediated transcriptional response of cultured urothelial cells under conditions of PHD inhibition. Finally, Aim 3 will evaluate the integral role of urothelial-cell specific HIF expression during injury-induced cystitis.