Cellular and Molecular Basis of Sex Specificity in UTI Pathogenesis

UTI 发病机制中性别特异性的细胞和分子基础

• 批准号: 10264141
• 负责人: Ashlee Miriam Earl
• 金额: $ 47.25万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10264141
• 关键词: Acute; Affect; Anatomy; Androgen Receptor; Androgenization; Androgens; Antibiotics; Automobile Driving; Bacteria; Bacterial Infections; Bacterial RNA; Biological Assay; Bladder; Bypass; Catheters; Cell Culture System; Cell Nucleus; Child; Chronic Cystitis; Chronic Kidney Failure; Cicatrix; Collaborations; Communities; Complement Factor B; Complex; Complication; Data; Epidemiology; Epithelial Cells; Escherichia coli Infections; Exhibits; Female; Flow Cytometry; Functional disorder; Gastrointestinal tract structure; Gene Expression; Genetic Transcription; Gonadal Steroid Hormones; Health; Healthcare; Histology; Hormonal; Human; Hybrids; Hypertension; Immune response; In Vitro; Inbred C3H Mice; Incidence; Infection; Infective cystitis; Kidney; Knock-out; Knockout Mice; Laboratories; Leukocytes; Life; Link; Mediator of activation protein; Methods; Microscopy; Modeling; Molecular; Morbidity - disease rate; Mouse Strains; Mus; Myofibroblast; Operative Surgical Procedures; Outcome; Pathogenesis; Pathogenicity; Pathway interactions; Phenotype; Polycystic Ovary Syndrome; Population; Pre-Clinical Model; Predisposition; Publishing; Pyelonephritis; Recurrence; Resolution; Risk; SHH gene; Severities; Sex Differences; Signal Pathway; Signal Transduction; Site; Specific qualifier value; Specificity; Techniques; Technology; Testing; Time; To specify; Transforming Growth Factors; Tubular formation; Universities; Urethra; Urinary tract; Urinary tract infection; Uropathogenic E. coli; Vesico-Ureteral Reflux; Virulence; Virulence Factors; Washington; Woman; Work; activin A; cell type; conditional knockout; epidemiologic data; experimental study; in vivo; infection management; infection risk; innovation; insight; kidney fibrosis; kidney infection; macrophage; male; men; minimally invasive; mortality; mutant; novel; pathogen; patient population; pre-clinical; prevent; programs; recruit; renal abscess; renal epithelium; renal scarring; response; sex; transcriptome sequencing; young adult

PROJECT SUMMARY In the proposed work, the Hunstad laboratory at Washington University will employ new models of urinary tract infection (UTI) in female, androgenized female, and male mice to determine molecular mechanisms by which host androgen exposure promotes uropathogenic Escherichia coli (UPEC) pyelonephritis as well as renal scarring, a common sequela of upper-tract UTI. Our recent findings indicate that the influence of sex, including androgen exposure, on these common bacterial infections is more complex than previously appreciated. Using newly developed and optimized models of UTI in mice, we recently demonstrated that androgen exposure is associated with increased risk for chronic cystitis and high-titer pyelonephritis, as well as formation of renal abscesses, in both male and female hosts. These findings correlate with epidemiologic data revealing higher morbidity and mortality in men who do suffer complicated UTI (compared with women), and higher incidence of UTI in women with a common hyperandrogenic condition (polycystic ovary syndrome). Moreover, we have demonstrated a separable effect of androgens on renal fibrosis (scarring), a common complication of pyelonephritis in children that can contribute to long-term sequelae such as hypertension and risk for chronic kidney disease. On the basis of these findings, we hypothesize that fundamental sex differences impact the host-pathogen interaction and cellular responses in pyelonephritis, thereby influencing pathogenicity, resolution, and subsequent renal scarring. To interrogate this hypothesis, and building on our published work, we will first comprehensively define host-sex-specific virulence requirements and sex influences on the host- pathogen interaction through single-nucleus RNA-seq, hybrid capture-enhanced bacterial RNA-seq, and insertion-site sequencing experiments on the infected kidney, in collaboration with MPI Dr. Earl and collaborator Dr. Humphreys. Candidate sex-discrepant host pathways and bacterial virulence factors will be confirmed and interrogated using germline and conditional knockout mice, bacterial mutants, and an array of bacterial pathogenesis studies. We will also define the molecular pathways underlying renal fibrosis following pyelonephritis, and how these pathways are modulated by sex and/or by androgen exposure at varying times before, during, and after UTI. In total, the proposed work will leverage new preclinical models and an array of conventional and cutting-edge experimental techniques. Our results will illuminate sex-specific host-pathogen interactions in the infected kidney and identify UPEC virulence factors important in sex-dependent outcomes of upper-tract UTI. In addition, our preclinical findings will also be translationally relevant to recurrent UTI and renal scarring in human patient populations.

项目摘要 在拟议的工作中，华盛顿大学的Hunstad实验室将采用新的尿路模型 雌性，雄性雌性和雄性小鼠的感染（UTI）确定分子机制 宿主雄激素暴露促进肝病大肠杆菌（UPEC）肾盂肾炎和肾脏 疤痕，上提取uti的常见后遗症。我们最近的发现表明，性的影响，包括 在这些常见的细菌感染上，雄激素暴露比以前所欣赏的要复杂。 我们使用新成立和优化的UTI模型在小鼠中，我们最近证明了雄激素 暴露与增加慢性膀胱炎和高点肾盂肾炎的风险增加有关 男性和雌性宿主的肾脓肿。这些发现与流行病学数据揭示 遭受复杂UTI（与女性相比）的男性发病率和死亡率更高，并且更高 具有常见性雌激素状况（多囊卵巢综合征）的女性UTI的发生率。而且， 我们已经证明了雄激素对肾纤维化（疤痕）的可分离作用，这是一种常见的并发症 儿童的肾盂肾炎，可以导致长期后遗症，例如高血压和慢性风险 肾脏疾病。根据这些发现，我们假设基本的性别差异影响 肾盂肾炎中的宿主 - 病原体相互作用和细胞反应，从而影响致病性， 分辨率和随后的肾脏疤痕。询问这一假设并基于我们发表的工作， 我们首先将全面定义宿主特定的毒力要求和对宿主的性影响 - 通过单核RNA-Seq，杂化捕获增强细菌RNA-Seq和 与MPI伯爵博士合作，在感染肾脏上进行插入站点测序实验 合作者Humphreys博士。候选性别裁员宿主途径和细菌毒力因素将是 使用种系和有条件的敲除小鼠，细菌突变体和一系列阵列确认并审问 细菌发病机理研究。我们还将定义肾脏纤维化的分子途径 肾盂肾炎，以及这些途径如何通过性别和/或在不同时间暴露于雄激素的调节 在UTI之前，期间和之后。总的来说，拟议的工作将利用新的临床前模型和一系列 常规和尖端的实验技术。我们的结果将阐明特定性别的宿主病原 感染肾脏中的相互作用并确定在性别依赖性结果中重要的UPEC毒力因素 上提取的UTI。此外，我们的临床前发现也将在翻译上与经常性的UTI和 人类患者人群中的肾脏疤痕。