Elucidating the Role of Biofilm-Forming Bacteria in Nephrolithiasis

标题：阐明生物膜形成细菌在肾结石中的作用 项目总结/摘要 肾结石是常见的，影响11%的美国人口。但 肾结石的发病机制仍不清楚。虽然已知10%的石头 感染性钙结石占结石的80%，以前没有发现 与细菌相联系。然而，最近的出版物已经利用下一代测序技术， 证明钙结石中存在细菌感染，尽管没有证据表明先前或同时存在 尿路感染的患者。在前期工作中，我们发现尿路致病菌 与对照组相比，触发草酸钙晶体生长增加。我们假设 细菌与钙结石相互作用并促进肾结石的繁殖。我们进一步 提出这种相互作用涉及细菌生物膜的形成。 我们将使用三个具体目标来测试这些假设： 1)检验尿路病原体启动结石生物膜形成的假设 2)为了验证假单胞菌已知的表面传感途径。和大肠杆菌 引发肾结石形成， 3)为了验证细菌在触发结晶过程中起重要作用的假设， 使用体外三维肾在尿特异性环境中的晶体聚集 模型 这些目标将通过利用多种成像和互补技术来实现 包括拉曼光谱和X射线荧光以及单细胞跟踪分析 和分子遗传学技术来研究生物膜的起始。 这项研究将为细菌在钙基细菌繁殖中的作用提供新的见解。 肾结石这也将促进我的独立生物医学研究员的进步 事业我将与非常成功的人合作，并获得他们的指导和监护， 经验丰富的导师致力于我的专业发展。我的研究 能力将通过微生物学、机械和生物技术方面的教学和实践培训得到增强。 学习，和高分辨率成像，从而为我准备了过渡到 完全独立的外科医生兼科学家最后，这笔资金将使我能够为 阐明肾结石的病因，将直接为肾结石的治疗提供初步资料。 我的第一个R 01应用程序。

TITLE: Elucidating the Role of Biofilm-Forming Bacteria in Nephrolithiasis PROJECT SUMMARY/ ABSTRACT Kidney stone disease is common, affecting 11% of the US population. However, the pathogenesis of kidney stones is still not well understood. While 10% of stones are known to be infectious, calcium-based stones, which make up 80% of stones, have not previously been shown to associate with bacteria. However, recent publications have utilized next-generation sequencing to demonstrate bacterial infestation in calcium stones despite no evidence of prior or concurrent urinary tract infection in those patients. In preliminary work, we found that uropathogenic bacteria trigger increased calcium oxalate crystal growth compared to controls. We hypothesize that bacteria interact with calcium stones and promote the propagation of kidney stones. We further propose that this interaction involves the formation of bacterial biofilm. We will test these hypotheses using three specific aims: 1) To test the hypothesis that uropathogens initiate biofilm formation in stones 2) To test the hypothesis that known surface sensing pathways of Pseudomonas spp. and E coli trigger kidney stone formation and 3) To test the hypothesis that bacteria play an important role in triggering crystallization and crystal aggregation in urine-specific environments using an in vitro three-dimensional kidney model. These aims will be accomplished by utilizing multiple imaging and complementary techniques including Raman spectroscopy and x-ray fluorescence as well as single-cell tracking analyses and molecular genetic techniques to investigate biofilm initiation. This study will provide new insights into the role of bacteria in the propagation of calcium-based kidney stones. It will also facilitate the advancement of my independent biomedical researcher career. I will collaborate with and obtain guidance and tutelage from highly successful, experienced mentors who are committed to my professional development. My research capabilities will be augmented by didactic and hands-on training in microbiology, machine learning, and high-resolution imaging, thus preparing me with the tools necessary to transition to full independence as a surgeon-scientist. Lastly, this funding will allow me to contribute to elucidating the etiology of nephrolithiasis and will directly enable provide the preliminary data for my first R01 application.