Mechanisms of Cellular Adaptation in Cystitis

 描述（由申请人提供）：以下建议概述了我成为一名独立研究人员的计划，专注于为膀胱炎症患者寻找解决方案。我是范德比尔特大学泌尿外科系的助理教授，接受过儿科泌尿学的奖学金培训。我将把未来5年的职业生涯的首要目标是发展科学和专业能力，成为一个独立的科学家。膀胱炎或膀胱炎症是一种普遍的问题，每年影响数百万美国人，这是尿路感染、膀胱梗阻、放射和化疗的结果。膀胱炎引起膀胱损伤的患者会出现尿失禁、膀胱疼痛、血尿甚至肾损伤。许多膀胱炎症患者最终需要进行侵入性膀胱手术。一个基本的知识差距存在于理解膀胱的尿路上皮细胞如何适应损伤后，并在随后的炎症反应。增加对这个问题的了解对于开发治疗方法以减轻膀胱炎引起的短期和长期损害非常重要。我发展成为一名独立的研究人员将使我能够提出重要的科学问题，然后寻找答案，以改善膀胱炎症患者的生活。我将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，研究生院课程，技术培训，智力互动，校园研讨会，赠款写作课程和实验室会议。导师制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。我所在的机构拥有多种资源来促进职业发展，我将充分利用这些机会。新出现的证据表明，急性激活缺氧诱导因子（HIF）途径是有益的上皮细胞在损伤和炎症的设置。我们生成的初步数据表明，损伤后膀胱中发生了HIF激活。释放HIF信号通路潜在益处的关键在于含脯氨酰-4-羟化酶结构域（PHD）蛋白。PHDs是上游氧敏感酶，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活性导致HIF通路的激活，随后促进上皮存活和保护。我们已经证明，PHD抑制在尿路上皮损伤和膀胱炎的情况下是保护性的。根据我们的初步数据和其他器官的已发表数据，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，PHD抑制是这一过程中关键的第一步。我们将通过执行三个具体目标来检验中心假设。目的1试图了解如何抑制个别PHD 2家族成员保护膀胱免受损伤使用小鼠模型的膀胱炎。目的2探讨PHD抑制条件下HIF介导的尿路上皮细胞转录反应。最后，目标3将评估损伤诱导的膀胱炎中尿路上皮细胞特异性HIF表达的整体作用。

 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.