A Microbial Model for the Formation of Calcium Oxalate and Calcium Phosphate Stones

草酸钙和磷酸钙结石形成的微生物模型

基本信息

批准号：
10447853
负责人：
Qunfeng Dong
金额：
$ 20.6万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-25 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10447853
关键词：
16S ribosomal RNA sequencing Adherence Affect Bacteria Bacterial Genes Binding Biological Calcium Oxalate Coupled Crystal Formation Crystallization DNA sequencing Deposition Development Disease Economic Burden Escherichia coli Evaluation Excision Failure Flagella Functional disorder Gene Deletion Genes Genetic Genome Goals Growth In Vitro Kidney Kidney Calculi Knowledge Libraries Link Lipopolysaccharides Medical Membrane Methodology Methods Microbe Microbial Biofilms Modeling Mus Other Genetics Patients Persons Pilot Projects Pilum Population Prevalence Process Publishing Pyelonephritis Recurrence Research Risk Risk Factors Role Surface Technology Testing United States Urease Urinary Calculi Urinary tract Urinary tract infection Urine Vesicle biomineralization calcium phosphate capsule in vitro Assay in vivo in vivo evaluation metagenomic sequencing microbial microbiota mouse model mutant prognosis biomarker promoter renal calcium risk prediction struvite targeted treatment treatment strategy urinary

16S ribosomal RNA sequencing Adherence Affect Bacteria Bacterial Genes Binding Biological Calcium Oxalate Coupled Crystal Formation Crystallization DNA sequencing Deposition Development Disease Economic Burden Escherichia coli Evaluation Excision Failure Flagella Functional disorder Gene Deletion Genes Genetic Genome Goals Growth In Vitro Kidney Kidney Calculi Knowledge Libraries Link Lipopolysaccharides Medical Membrane Methodology Methods Microbe Microbial Biofilms Modeling Mus Other Genetics Patients Persons Pilot Projects Pilum Population Prevalence Process Publishing Pyelonephritis Recurrence Research Risk Risk Factors Role Surface Technology Testing United States Urease Urinary Calculi Urinary tract Urinary tract infection Urine Vesicle biomineralization calcium phosphate capsule in vitro Assay in vivo in vivo evaluation metagenomic sequencing microbial microbiota mouse model mutant prognosis biomarker promoter renal calcium risk prediction struvite targeted treatment treatment strategy urinary

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项目摘要

Project Summary/Abstract Urinary stones occur in 10% of people. The economic burden from urinary stones accounts for more than 2 billion dollars annually. Past studies have demonstrated that bacteria can be cultured from about one-third of urinary stones and we have provided the initial demonstration that DNA sequencing coupled with enhanced culture methods can be used to identify the urinary stone microbiota (the bacteria in urinary stones) in many more stones than by standard culturing alone. Yet, critical knowledge gaps exist concerning the underlying mechanisms of urinary stone pathophysiology. Alarmingly, prevalence of urinary stones is increasing, and new treatments are not being developed. Although urine supersaturated with calcium oxalate has been associated with stone formation, supersaturated urine is often present in non-stone formers. Accumulating evidence supports a bacterial role in urinary stone pathophysiology. First, bacteria aggregate to calcium oxalate (CaOx), the type of crystal responsible for most urinary stone disease. Second, initial in vitro studies indicate that bacterial flagella are important components in bacterial promotion of crystal aggregation. Third, bacteria increase the size of crystal clusters in an in vivo murine model. Our long-term research goal is to develop new urinary stone treatment strategies that reduce urinary stone recurrence. We hypothesize that flagella directly contribute to lithogenesis. To begin to determine the functional role of flagella in stone formation, we will test bacterial mutants lacking key flagella components and/or function for their ability to promote CaOx aggregation in vitro and in vivo. (Aim 1). Besides flagella, we also hypothesize that other genetic factors may contribute to lithogenesis as well. We will perform a bacteria mutant screen to identify non-flagellar genes involved in interactions with CaOx crystals. In addition, we will explore whether the identified genes are enriched in bacterial isolates associated with stones relative to urinary isolates not associated with stones (Aim 2).

项目摘要/摘要尿石发生在10％的人中。尿石的经济负担超过2个每年十亿美元。过去的研究表明，细菌可以从大约三分之一的尿石，我们提供了初始证明，即DNA测序与增强培养方法可用于鉴定许多尿石微生物群（尿石中的细菌）与单独培养相比，石头更多。然而，存在有关基础的关键知识差距尿石病理生理学的机制。令人震惊的是，尿石的患病率正在增加，并且没有开发新的治疗方法。尽管用草酸钙过饱和尿液已经与石材的形成相关，通常存在于非石材形成器中过饱和的尿液。累积证据支持在尿石病理生理学中的细菌作用。首先，细菌聚集至草酸钙（CAOX），这种晶体的类型负责大多数尿石疾病。第二，初始体外研究表明细菌鞭毛是细菌促进晶体聚集的重要组成部分。第三，细菌在体内鼠模型中增加晶体簇的大小。我们的长期研究目标是开发新的减少尿石复发的尿石处理策略。我们假设鞭毛直接有助于岩性生成。开始确定的功能作用石头形成的鞭毛，我们将测试缺乏关键鞭毛成分和/或功能的细菌突变体在体外和体内促进CAOX聚集的能力。（目标1）。除了鞭毛，我们还假设其他遗传因素也可能有助于岩性生成。我们将执行细菌突变筛查鉴定与CAOX晶体相互作用的非鞭毛基因。此外，我们将探索是否鉴定的基因富含与尿液分离株相关的结石相关的细菌分离株与石头相关（AIM 2）。