Molecular and Cellular Pathobiology of Stone Forming Papillae

在上一个供资周期，项目小组在我们对以下机制的理解方面取得了重大进展 石块的形成和生长。项目3已经做了全面和必要的组织病理学 人类结石病中乳头的特征，并区分了两个主要表型，即 对结石疾病的发病机制至关重要：斑块沉积和肾小管内晶体堵塞。对…的把握 这些表型中的每一种对结石疾病类型的贡献对于理解 基础病理学，并最终开发有针对性的治疗方法。实现这一目标需要 对人类乳头内各种微环境进行了前所未有的、详细的研究 形成结石的病人。当前提案中的目标旨在执行这种深入的 使用最先进的技术研究这些关键事件背后的细胞和分子机制 研究团队的方法和技能。 间质钙浓度在斑块形成中的作用是一个基本概念。目标1将 牢固确立乳头间质钙增多在斑块发病机制中的重要性。 目的2探讨先天免疫激活在斑块形成中的作用。的关键下游影响 先天免疫，如氧化应激、炎症信号和晶体调节剂沉积 仔细研究过。预计这一目标将高度自信地定义炎症在疾病中的作用 人类结石病中斑块沉积的发病机制。 目标3将研究堵塞疾病如何在乳头中传播的关键机制，通过确定 带有塞子的管道可以刺激邻近区域以外的氧化应激和炎症信号 堵住了。这种串扰可能导致黏附分子的上调，从而促进更多的插入 相邻的管道。最后，目标4将研究很少研究的乳头状尿路上皮，通过建立 新结石在斑块上过度生长之前，其上覆的乳头状尿路上皮功能障碍。 在这些研究中使用的方法包括：激光显微解剖，乳头的三维成像 使用共聚焦显微镜、组织细胞术、最新的蛋白质分析和氧化还原蛋白质组分析。 这些新的目标结合在一起，并与项目1和2中的目标协同，将极大地推动我们的 对结石形成和生长的精确机制的了解，这有望转化为 对结石病的临床治疗更加有效。

During the last funding cycle, the PPG made significant advances in our understanding of the mechanisms of stone formation and growth. Project 3 has made a comprehensive and essential histopathological characterization of the papillae in human stone disease, and distinguished two main phenotypes that are essential to the pathogenesis of stone disease: plaque deposition and intratubular crystal plugging. A grasp of the contribution of each of these phenotypes to the type of stone disease is essential to understand the underlying pathology, and eventually developing targeted therapies. Achieving this objective requires the performance of unprecedented, detailed, studies of the various microenvironments within the human papillae of stone forming patients. The aims in the current proposal are designed to perform such in-depth investigations of the cellular and molecular mechanisms underlying these key events, using state-of-the-art methodology and skills of the research team. The role of interstitial calcium concentration in the formation of plaque is a fundamental concept. Aim 1 will firmly establish the importance of increased interstitial calcium in the papilla on the pathogenesis of plaque. Aim 2 will investigate the role of innate immune activation in plaque formation. Key downstream effects of innate immunity, such as oxidative stress, inflammatory signaling and crystal modulator deposition will be carefully studied. This aim is expected to define with high confidence, the role of inflammation in the pathogenesis of plaque deposition in human stone disease. Aim 3 will investigate a key mechanism of how plugging disease propagates in the papilla, by determining if ducts with plugs can stimulate oxidative stress and inflammatory signaling beyond the area contiguous to plugging. This cross -talk could lead to upregulation of adhesion molecules that facilitate more plugging in neighboring ducts. Finally, Aim 4 will investigate the rarely-studied papillary urothelium, by establishing if overgrowth of new stones over plaque is preceded by dysfunction of the overlying papillary urothelium. Methods used in these investigations will include: laser micro-dissection, 3 dimensional imaging of the papillae using confocal microscopy, tissue cytometry, state-of the art protein analysis and redox proteomic assays. Combined together, and in synergy with aims from project 1 and 2, these new aims, will greatly advance our understanding of the precise mechanisms of stone formation and growth, which will hopefully translate into more effective clinical treatments for stone disease.